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WO2025006499A2 - Gpc3-targeted molecules and uses thereof - Google Patents

Gpc3-targeted molecules and uses thereof Download PDF

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Publication number
WO2025006499A2
WO2025006499A2 PCT/US2024/035468 US2024035468W WO2025006499A2 WO 2025006499 A2 WO2025006499 A2 WO 2025006499A2 US 2024035468 W US2024035468 W US 2024035468W WO 2025006499 A2 WO2025006499 A2 WO 2025006499A2
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Prior art keywords
seq
amino acid
acid sequence
set forth
sequence set
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French (fr)
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WO2025006499A3 (en
Inventor
Xianhui Chen
Cokey Nguyen
Michel Sadelain
Leena HALIM
Michael Lopez
Shuai Yang
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Memorial Sloan Kettering Cancer Center
Atara Biotherapeutics Inc
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Memorial Sloan Kettering Cancer Center
Atara Biotherapeutics Inc
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Publication of WO2025006499A2 publication Critical patent/WO2025006499A2/en
Publication of WO2025006499A3 publication Critical patent/WO2025006499A3/en
Anticipated expiration legal-status Critical
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A61K40/4261—Proteoglycans, e.g. glypican, brevican or CSPG4
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/10—Cellular immunotherapy characterised by the cell type used
    • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
    • A61K40/31—Chimeric antigen receptors [CAR]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/303—Liver or Pancreas
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2239/00—Indexing codes associated with cellular immunotherapy of group A61K40/00
    • A61K2239/46—Indexing codes associated with cellular immunotherapy of group A61K40/00 characterised by the cancer treated
    • A61K2239/53—Liver
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • TECHNICAL FIELD [0001] The presently disclosed subject matter provides molecules that target glypican-3 (GPC3), cells comprising the molecules, and methods for treating GPC3-related diseases using such molecules and cells.
  • GPC3 glypican-3
  • Cell-based immunotherapy is a therapy with curative potential for the treatment of cancer. T-cells and other immune cells may be modified to target tumor antigens through the introduction of genetic material coding for artificial or synthetic receptors for antigen, termed Chimeric Antigen Receptors (CARs), specific to selected antigens. Targeted T-cell therapy using CARs has shown recent clinical success in treating hematologic malignancies.
  • CARs Chimeric Antigen Receptors
  • GPC3 is widely expressed in the placenta and fetal tissues, while its expression is significantly reduced in adult organs.
  • significantly high expression levels of GPC3 have been found in hepatocellular carcinoma (HCC) cells compared to normal liver and non-neoplastic liver lesions.
  • HCC hepatocellular carcinoma
  • GPC3-targeted molecules e.g., CARs or antibodies, for treating tumors, e.g., HCC. 3.
  • the GPC3-targeted CAR comprises an extracellular domain that binds to GPC3, a transmembrane domain, and an intracellular domain, wherein the extracellular domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: (a) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, and the V L comprises a V L CDR1, a V L CDR2, and a V L CDR3 of a V L comprising the amino acid sequence set forth in SEQ ID NO: 30; (b) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 47, and the V L comprises a V L CDR1, a V L CDR2, and a V L CDR3 of a V L comprising
  • the V H CDR1, V H CDR2, V H CDR3, V L CDR1, V L CDR2, and V L CDR3 are identified according to the Kabat numbering system, the Chothia numbering system, the AbM numbering system, the Contact numbering system, or the IMGT ® Information System. In certain embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system.
  • the VH and VL are characterized below: (a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative NAI-1540294631v3 3 modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof; (b) the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a
  • the VH and VL are characterized below: (a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28; NAI-1540294631v3 7 (b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the V L comprises a CDR1 comprising the amino acid sequence set forth
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the V L NAI-1540294631v3 10 comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
  • the VH comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262.
  • the VL comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263.
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263.
  • the V H and V L are characterized below: (a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; (b) the V H comprises the amino acid sequence set forth in SEQ ID NO: 47; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 48; (c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 62; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 63; NAI-1540294631v3 11 (d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; (e) the V H comprises the amino acid sequence set forth in SEQ ID NO: 89; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 90; (f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 101;
  • the VH and VL are characterized below: (a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; (b) the V H comprises the amino acid sequence set forth in SEQ ID NO: 47; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 48; (c) the V H comprises the amino acid sequence set forth in SEQ ID NO: 75; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 76; NAI-1540294631v3 12 (d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; (e) the V H comprises the amino acid sequence set forth in SEQ ID NO: 155; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 156; or (f) the VH comprises the amino acid sequence set forth in SEQ ID NO:
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 262; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 263.
  • the VH and the VL are connected via a linker.
  • the linker consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.
  • the linker consists of the amino acid sequence set forth in SEQ ID NO: 1.
  • the VH and the VL are positioned from the N- to the C-terminus: V L -V H . In certain embodiments, the V H and the V L are positioned from the N- to the C-terminus: V H -V L .
  • the extracellular domain comprises a variable fragment (Fv), a single-chain variable fragment (scFv), a Fab, or a F(ab) 2 . In certain embodiments, the extracellular domain comprises an scFv.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO:
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 264, or SEQ ID NO: 265.
  • the transmembrane domain comprises a CD8 polypeptide, a CD28 polypeptide, a CD3 ⁇ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3 NAI-1540294631v3 13 polypeptide, a 2B4 polypeptide, or a BTLA polypeptide.
  • the transmembrane domain comprises a CD28 polypeptide.
  • the CD28 polypeptide comprises amino acids 154 to 179 of the amino acid sequence set forth in SEQ ID NO: 308.
  • the intracellular domain comprises a CD3 ⁇ polypeptide.
  • the CD3 ⁇ polypeptide is a modified CD3 ⁇ polypeptide.
  • the modified CD3 ⁇ polypeptide comprises a native ITAM1, an ITAM2 variant consisting of two loss-of-function mutations, and an ITAM3 consisting of two loss-of-function mutations.
  • the native ITAM1 consists of the amino acid sequence set forth in SEQ ID NO: 313.
  • the ITAM2 variant consists of the amino acid sequence set forth in SEQ ID NO: 319.
  • the ITAM3 variant consists of the amino acid sequence set forth in SEQ ID NO: 323.
  • the modified CD3 ⁇ polypeptide comprises or consists of the amino acid sequence set forth in SEQ ID NO: 325.
  • the intracellular domain further comprises at least one co-stimulatory signaling region.
  • the at least one co-stimulatory signaling region comprises at least one intracellular signaling domain of a co-stimulatory molecule.
  • the co- stimulatory molecule is selected from the group consisting of CD28, 4-1BB, OX40, ICOS, DAP-10, CD27, CD28, CD30, CD40, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, CD8, CD4, B2C, CD80, CD86, DAP10, DAP12, MyD88, BTNL3, and NKG2D, and combinations thereof.
  • the co-stimulatory molecule is CD28.
  • the at least one co-stimulatory signaling region comprises amino acids 180 to 220 of the amino acid sequence set forth in SEQ ID NO: 308.
  • the CAR further comprises a signal peptide.
  • the signal peptide comprises the amino acid sequence set forth in SEQ ID NO: 278.
  • the CAR further comprises a self-cleaving P2A peptide.
  • the self-cleaving P2A peptide comprises the amino acid sequence set forth in SEQ ID NO: 329.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 333, SEQ ID NO: 335, SEQ ID NO: 337, SEQ ID NO: 339, SEQ ID NO: 341, SEQ ID NO: 343, SEQ ID NO: 345, SEQ ID NO: 347, SEQ ID NO: 349, SEQ ID NO: 351, SEQ ID NO: 353, or SEQ ID NO: 355.
  • the CAR is expressed from a vector.
  • the vector is a viral vector.
  • the viral vector is a retroviral vector.
  • the presently disclosed subject matter further provides nucleic acids encoding the GPC3- targeted CARs disclosed herein.
  • the nucleic acid further comprises a promoter that is operably linked to the CAR.
  • the promoter is endogenous or exogenous.
  • the exogenous promoter is selected from the group consisting of an elongation factor (EF)-1 promoter, a cytomegalovirus immediate-early promoter (CMV) promoter, a simian virus 40 early promoter (SV40) promoter, a phosphoglycerate kinase (PGK) promoter, a metallothionein promoter, and Ubiquitin C promoter.
  • the endogenous promoter is selected from the group consisting of a TCR alpha promoter, a TCR beta promoter, and a beta 2-microglobulin promoter.
  • the promoter is an inducible promoter.
  • the inducible promoter is selected from the group consisting of a NFAT transcriptional response element (TRE) promoter, a CD69 promoter, a CD25 promoter, an IL-2 promoter, a 4-1BB promoter, a PD1 promoter, and a LAG3 promoter.
  • TRE NFAT transcriptional response element
  • the nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO: 334, SEQ ID NO: 336, SEQ ID NO: 338, SEQ ID NO: 340, SEQ ID NO: 342, SEQ ID NO: 344, SEQ ID NO: 346, SEQ ID NO: 348, SEQ ID NO: 350, SEQ ID NO: 352, SEQ ID NO: 354, or SEQ ID NO: 356.
  • the presently disclosed subject matter provides vectors comprising the nucleic acids disclosed herein.
  • the vector is a viral vector.
  • the viral vector is a retroviral vector.
  • the presently disclosed subject matter further provides cells comprising the CARs, nucleic acids, or vectors disclosed herein.
  • the cell is an immunoresponsive cell or an immune effector cell.
  • the cell is a cell of the lymphoid lineage or a cell of the myeloid lineage.
  • the cell is selected from the group consisting of a T cell, a B cell, a Natural Killer (NK) cell, macrophage, an innate lymphoid cell (ILC), a cytokine induced killer (CIK) cell, a lymphokine activated killer (LAK) cell, a stem cell from which a lymphoid cell may be differentiated, a stem cell from which a myeloid cell may be differentiated, or a combination thereof.
  • the cell is a T cell.
  • the T cell is selected from the group consisting of helper T cells, cytotoxic T cells, memory T cells, regulatory T cells, tumor- infiltrating lymphocyte (TIL), Natural Killer T cells, mucosal associated invariant T cells, ⁇ T cells, and ⁇ T cells.
  • the cell is a NK cell.
  • the NK cell is NAI-1540294631v3 15 derived from a stem cell.
  • the stem cell is a pluripotent stem cell.
  • the pluripotent stem cell is an embryoid stem cell or an induced pluripotent stem cell.
  • the presently disclosed subject matter provides anti-GPC3 antibodies or antigen-binding fragments thereof.
  • the anti-GPC3 antibody or antigen- binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (V L ), wherein: (a) the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 of a V H comprising the amino acid sequence set forth in SEQ ID NO: 29, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a V L comprising the amino acid sequence set forth in SEQ ID NO: 30; (b) the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 of a V H comprising the amino acid sequence set forth in SEQ ID NO: 47, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising
  • the V H CDR1, V H CDR2, V H CDR3, V L CDR1, V L CDR2, and V L CDR3 are identified according to the Kabat numbering system, the Chothia numbering system, the AbM numbering system, the Contact numbering system, or the IMGT ® Information System. In certain embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system.
  • the VH and VL are characterized as below: (a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative NAI-1540294631v3 17 modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof; (b) the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in
  • the VH and VL are characterized as below: (a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28; NAI-1540294631v3 21 (b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the V L comprises a CDR1 comprising the amino acid sequence set
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the V L NAI-1540294631v3 24 comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
  • the VH comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262.
  • the VL comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263.
  • the V H and V L are characterized as below: (a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; (b) the V H comprises the amino acid sequence set forth in SEQ ID NO: 47; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 48; (c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 62; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 63; NAI-1540294631v3 25 (d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; (e) the V H comprises the amino acid sequence set forth in SEQ ID NO: 89; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 90; (f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 101
  • the VH and VL are characterized as below: (a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; (b) the V H comprises the amino acid sequence set forth in SEQ ID NO: 47; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 48; (c) the V H comprises the amino acid sequence set forth in SEQ ID NO: 75; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 76; NAI-1540294631v3 26 (d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; (e) the V H comprises the amino acid sequence set forth in SEQ ID NO: 155; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 156; or (f) the VH comprises the amino acid sequence set forth in SEQ ID NO:
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 262; and the V L comprises the amino acid sequence set forth in SEQ ID NO: 263.
  • the antigen-binding fragment thereof is a variable fragment (Fv), a single-chain variable fragment (scFv), a Fab, or a F(ab)2.
  • the antigen- binding fragment thereof is an scFv.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO:
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 264, or SEQ ID NO: 265.
  • the VH is connected to the VL via a linker.
  • the linker consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.
  • the linker consists of the amino acid sequence set forth in SEQ ID NO: 1.
  • the VH and the VL are positioned from the N- to the C-terminus: VL-VH.
  • the VH and the VL are positioned from the N- to the C-terminus: V H -V L .
  • the antibody comprises a human variable region framework region.
  • the antibody or antigen-binding fragment thereof comprises a Fc region.
  • the Fc region comprises the amino acid sequence set forth in SEQ ID NO: 357.
  • the antibody or antigen-binding fragment thereof is a fully human or an antigen-binding fragment thereof, a chimeric antibody or an antigen-binding fragment thereof, or a humanized antibody or an antigen-binding fragment thereof.
  • the presently disclosed subject matter further provides antibodies or antigen-binding fragments thereof that compete with any one of the anti-GPC3 antibodies or antigen-binding fragments thereof disclosed herein for binding to GPC3.
  • the presently disclosed subject matter provides antibodies or antigen-binding fragments thereof that bind to essentially the same epitope region on GPC3 as any one of the anti-GPC3 antibodies or antigen-binding fragments thereof disclosed herein.
  • the presently disclosed subject matter further provides conjugates comprising the antibodies or antigen-binding fragments thereof disclosed herein.
  • the antibody or antigen-binding fragment thereof is linked to a therapeutic agent, a detectable agent, or a diagnostic agent.
  • the therapeutic agent is a chemotherapeutic agent, a cytotoxin, or a drug.
  • the conjugate is an immunoconjugate.
  • the presently disclosed subject matter provides multispecific molecules comprising any one of the antibodies or antigen-binding fragments thereof disclosed herein that is linked to a second functional moiety. In certain embodiments, the second functional moiety has a different binding specificity than the antibody or antigen binding fragment thereof.
  • the presently disclosed subject matter provides nucleic acids that encode the antibodies or antigen-binding fragments thereof.
  • the nucleic acid comprises a first polynucleotide that encodes a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262; and/or a second polynucleotide that encodes a VL comprising the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90,
  • the first polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO: 33, SEQ ID NO: 51, SEQ ID NO: 66, SEQ ID NO: 79, SEQ ID NO: 93, SEQ ID NO: 105, SEQ ID NO: 123, SEQ ID NO: 141, SEQ ID NO: 159, SEQ ID NO: 174, SEQ ID NO: 187, SEQ ID NO: 205, SEQ ID NO: 219, SEQ ID NO: 237, SEQ ID NO: 250, or SEQ ID NO: 266.
  • the second polynucleotide comprises the nucleotide sequence set forth in NAI-1540294631v3 28 SEQ ID NO: 35, SEQ ID NO: 53, SEQ ID NO: 68, SEQ ID NO: 81, SEQ ID NO: 95, SEQ ID NO: 107, SEQ ID NO: 125, SEQ ID NO: 143, SEQ ID NO: 161, SEQ ID NO: 176, SEQ ID NO: 189, SEQ ID NO: 207, SEQ ID NO: 221, SEQ ID NO: 239, SEQ ID NO: 252, or SEQ ID NO: 268.
  • the presently disclosed subject matter provides vectors comprising the nucleic acids disclosed herein.
  • the vector is an expression vector.
  • the presently disclosed subject matter further provides host cells comprising the vectors disclosed herein.
  • the presently disclosed subject matter provides methods of producing a presently disclosed anti-GPC3 antibody or an antigen-binding fragment thereof.
  • the method comprises culturing a presently disclosed host cell under conditions to induce expression of the antibody or antigen-binding fragment thereof from the host cell.
  • the presently disclosed subject matter provides methods for detecting GPC3 in a whole cell or tissue.
  • the method comprises: (a) contacting a cell or tissue with a presently disclosed anti-GPC3 antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises a detectable label; and (b) determining the amount of the labeled antibody or antigen-binding fragment thereof bound to the cell or tissue by measuring the amount of detectable label associated with the cell or tissue, wherein the amount of bound antibody or antigen-binding fragment thereof indicates the amount of GPC3 in the cell or tissue.
  • the presently disclosed composition comprises the cells, the antibodies or antigen- binding fragments, the conjugates, or the multispecific molecules disclosed herein.
  • the composition is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.
  • the composition comprises between about 1 ⁇ 10 5 and about 5 ⁇ 10 8 cells.
  • the presently disclosed cells, antibodies or antigen-binding fragments thereof, conjugates, multispecific molecules, or compositions can be used in a therapy.
  • the presently disclosed cells, antibodies or antigen-binding fragments thereof, conjugates, multispecific molecules, or compositions can be used in a method for treating a disease or disorder associated with GPC3 in a subject.
  • the presently disclosed subject matter provides methods of treating a disease or disorder associated with GPC3 in a subject.
  • the method comprises: administering to the subject cells comprising a presently disclosed GPC3-targeted CAR, the anti-GPC3 antibody or NAI-1540294631v3 29 antigen-binding fragment thereof disclosed herein, the conjugate disclosed herein, the multispecific molecule disclosed herein, or the composition disclosed herein.
  • the presently disclosed subject matter provides uses of the presently disclosed cells, antibodies or antigen-binding fragments thereof, conjugates, multispecific molecules, or compositions in the manufacture of a medicament for treating a disease or disorder associated with GPC3.
  • the disease or disorder is a tumor. In certain embodiments, the disease or disorder is cancer.
  • the disease or disorder is selected from the group consisting of hepatocellular carcinoma, hepatoblastoma, lung squamous cell carcinoma, ovarian yolk sac tumor, melanoma, and urothelial carcinoma. In certain embodiments, the disease or disorder is hepatocellular carcinoma. In certain embodiments, the subject is a human subject. [0062]
  • the presently disclosed subject matter provides methods for producing a cell comprising a presently disclosed GPC3-targeted CAR. In certain embodiments, the method comprises: introducing into a cell a nucleic acid that encodes the presently disclosed GPC3-targeted CAR. 4.
  • Figures 1A to 1E depict the binding activity of scFv-Fc fusion proteins of 16 binders to cells expressing human GPC3 using fluorescence-activated cell sorting (FACS) analysis.
  • FACS fluorescence-activated cell sorting
  • FIG. 1 An anti-hIgG GPC3 antibody was used as a positive control, and a human IgG was used as an isotype control, both of which were diluted in the same manner as the tested proteins.
  • a PBS solution was used as a negative control.
  • Each of the purified scFv-Fc fusion proteins was also tested for binding to a negative control cell line, CHO-K1(-). Plates were read by flow cytometry reader. Binding curves were plotted using the geometric-mean fluorescence intensity of the fluorescence signal on the cells, and the EC50 values were calculated.
  • Figures 2A and 2B depict the generation of the presently disclosed GPC3-targeted scFv CAR T Cells.
  • Figure 2A depicts structures of CARs in accordance with certain embodiments of the presently disclosed subject matter.
  • Figure 2B depicts the flow cytometric analysis showing the expression levels of LNGFR for the indicated constructs in the VH-VK and VK-VH orientation.
  • Figure 3 depicts the GPC3 expression levels of Nalm-6 GPC3 + , Huh7, and PLC/PRF/5 tumor cells.
  • Figures 4A and 4B depict the in vitro cytotoxicity of the presently disclosed GPC3- targeted CAR T cells against Nalm-6 GPC3 + and Nalm-6 WT tumor cells.
  • Figure 4A shows the NAI-1540294631v3 30 results of 16 hour Cytotoxic T lymphocyte (CTL) mediated cytotoxicity using Nalm-6 GPC3 + tumor cells.
  • Figure 4B shows the results of 16 hour CTL using Nalm-6 wildtype (WT) tumor cells.
  • Figure 5 depicts the in vitro cytotoxicity of the presently disclosed GPC3-targeted CAR T cells against Huh-7 tumor cells.
  • Figure 5 shows the results of 16 hour CTL using Huh-7 tumor cells.
  • Figure 6 depicts the in vitro cytotoxicity of the presently disclosed GPC3-targeted CAR T cells against PLC/PRF/5 tumor cells.
  • Figure 6 shows the results of 16 hour CTL using PLC/PRF/5 tumor cells.
  • Figure 7 depicts an experimental layout of an assessment of the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells.
  • Figure 8 depicts the body weight changes of mice who received the presently disclosed GPC3-targeted CAR T cells.
  • Figure 9 depicts the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells.
  • Figure 10 depicts an experimental layout of an assessment of the in vivo safety and effective dose of the presently disclosed GPC3-targeted CAR with binders 8 and 16, respectively.
  • Figure 11 depicts the survival rates of mice treated with the presently disclosed GPC3- targeted CAR with binders 8 and 16, respectively, at difference doses.
  • Figure 12 depicts body weight changes of mice treated with the presently disclosed GPC3-targeted CAR with binders 8 and 16, respectively, at difference doses.
  • Figure 13 depicts the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR with binders 8 and 16, respectively, at difference doses.
  • Figure 14 depicts the safety of the presently disclosed GPC3-targeted CAR with binders 8 and 16, respectively, at difference doses.
  • Figures 15A to 15C depict the restimulation assay with Huh7 tumor cells.
  • Figure 15A depicts a scheme showing the principle of CAR T restimulation assay.
  • Figure 15B shows the results of the Huh7 tumor cell lysis of the VH-VK GPC3 CAR T cells.
  • Figure 15C shows the results of the proliferation of the CAR T cells.
  • Figure 16 depicts an assessment of the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells to orthotopically implanted Huh7 tumor cells in liver.
  • Figure 17 depicts an assessment of the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells to subcutaneously implanted Huh7 tumor cells.
  • Figure 18 depicts an assessment of the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells to Nalm-6 GPC3+ tumor cells injected via tail vein. NAI-1540294631v3 31
  • Figures 19A and 19B depict an analysis of GPC3 expression level in tumor liver samples versus normal liver samples.
  • Figure 19A shows the density plot of upregulated membrane-bound proteins from mass spectrometry data obtained from the Chinese Human Proteome Project (CNHPP) (Jiang et al., 2019, Nature 567: 257–261) and the Clinical Proteomic Tumor Analysis Consortium (CPTAC) (Gao et al., 2019, Cell 179 (2): 561 - 577.e22), respectively. Proteins below the 98th percentile ranking of expression are colored in light color, and those in and above the 98th percentile ranking are colored in dark color.
  • CNHPP Chinese Human Proteome Project
  • CTAC Clinical Proteomic Tumor Analysis Consortium
  • Figure 19B shows the density plot of upregulated membrane- bound transcripts from bulk RNAseq data obtained from The Cancer Genome Atlas (TCGA) (Ally et al., 2017, Cell 169 (7): 1327 - 1341.e23) and the CPTAC. Transcripts below the 98th percentile ranking of expression are colored in light color, and those in and above the 98th percentile ranking are colored in dark color. 5.
  • TCGA Cancer Genome Atlas
  • TCGA Cancer Genome Atlas
  • the molecule is an antibody or an antigen- binding fragment thereof.
  • the molecule is an antigen recognizing receptor.
  • the antigen recognizing receptor is a chimeric antigen receptor (CAR).
  • CAR chimeric antigen receptor
  • the presently disclosed subject matter further discloses cells comprising such GPC3-targeted molecules.
  • the cells can be immunoresponsive cells and/or immune effector cells, e.g., genetically modified immunoresponsive cells (e.g., T cells or NK cells), or pluripotent stem cell that can differentiate into immunoresponsive cells and/or immune effector cells.
  • Non-limiting embodiments of the present disclosure are described by the present specification and Examples. [0084] For purposes of clarity of disclosure and not by way of limitation, the detailed description is divided into the following subsections: 5.1. Definitions; 5.2. GPC3; 5.3. Chimeric Antigen Receptors (CARs); 5.4.
  • immunoresponsive cell refers to a cell that functions in an immune response or a progenitor, or progeny thereof. In certain embodiments, the immunoresponsive cell is a cell of lymphoid lineage.
  • Non-limiting examples of cells of lymphoid lineage include T cells, Natural Killer (NK) cells, B cells, and stem cells from which lymphoid cells may be differentiated.
  • the immunoresponsive cell is a cell of myeloid lineage.
  • the term “activates an immunoresponsive cell” refers to induction of signal transduction or changes in protein expression in the cell resulting in initiation of an immune response. For example, when CD3 chains cluster in response to ligand binding and immunoreceptor tyrosine-based inhibition motifs (ITAMs) a signal transduction cascade is produced.
  • ITAMs immunoreceptor tyrosine-based inhibition motifs
  • an immunological synapse occurs that includes clustering of many molecules near the bound receptor (e.g., CD4 or CD8, CD3 ⁇ / ⁇ / ⁇ / ⁇ , etc.). This clustering of membrane bound signaling molecules allows for ITAM motifs contained within the CD3 chains to become phosphorylated. This phosphorylation in turn initiates a T cell activation pathway ultimately activating transcription factors, such as NF- ⁇ B and AP-1. These transcription factors induce global gene expression of the T cell to increase IL-2 production for proliferation and expression of master regulator T cell proteins in order to initiate a T cell mediated immune response.
  • CD4 or CD8 CD3 ⁇ / ⁇ / ⁇ / ⁇ , etc.
  • the term “stimulates an immunoresponsive cell” refers to a signal that results in a robust and sustained immune response. In certain embodiments, this occurs after NAI-1540294631v3 33 immunoresponsive cell (e.g., T cell) activation or concomitantly mediated through receptors including, but not limited to, CD28, 4-1BB, OX40, CD40 and ICOS. Receiving multiple stimulatory signals can be important to mount a robust and long-term T-cell mediated immune response. T cells can quickly become inhibited and unresponsive to antigen.
  • immunoresponsive cell e.g., T cell activation or concomitantly mediated through receptors including, but not limited to, CD28, 4-1BB, OX40, CD40 and ICOS.
  • antibody covers, for example polyclonal antibodies, monoclonal antibodies (including agonist, antagonist, neutralizing antibodies, full length monoclonal antibodies), antibody compositions with polyepitopic or monoepitopic specificity, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), synthetic antibodies, chimeric antibodies, humanized antibodies, or human versions of antibodies having full length heavy and/or light chains.
  • the present disclosure also includes antibody fragments (and/or polypeptides that comprise antibody fragments) that retain ⁇ 5 integrin binding characteristics.
  • antibody fragments include antigen-binding regions and/or effector regions of the antibody, e.g., Fab, Fab’, F(ab’) 2 , Fv, scFv, (scFv) 2 , single chain antibody molecule, dual variable region antibody, single variable region antibody, linear antibody, V region, a multispecific antibody formed from antibody fragments, F(ab)2, Fd, Fc, diabody, di-diabody, disulfide-linked Fvs (dsFv), single-domain antibody (e.g., VHH, nanobody) or other fragments (e.g., fragments consisting of the variable regions of the heavy and light chains that are non-covalently coupled).
  • variable region domain may be any suitable arrangement of immunoglobulin heavy (V H ) and/or light (V L ) variable regions.
  • V H immunoglobulin heavy
  • V L light
  • the presently disclosed subject matter also includes tetrameric antibodies comprising two heavy chain and two light chain molecules, an antibody light chain monomer, and an antibody heavy chain monomer.
  • the variable region domain may be dimeric and contain VH-VH, VH-VL, or VL-VL dimers that bind ⁇ 5 integrin.
  • the V H and V L chains may be covalently coupled either directly or through a linker to form a single chain Fv (scFv).
  • scFv proteins are referred to herein as included in the category “antibody fragments.”
  • Another form of an antibody fragment is a peptide comprising one or more complementarity determining regions (CDRs) of an antibody.
  • CDRs can be obtained by constructing polynucleotides that encode the CDR of interest.
  • Such polynucleotides are prepared, for example, by using the polymerase chain reaction to synthesize the variable region using mRNA of antibody-producing cells as a template (see, for example, Larrick et al., Methods: A Companion to Methods in Enzymology, 2:106 (1991); Courtenay-Luck, “Genetic Manipulation of NAI-1540294631v3 34 Monoclonal Antibodies,” in Monoclonal Antibodies Production, Engineering and Clinical Application, Ritter et al.
  • Antibody fragments may be incorporated into single domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, variable domains of new antigen receptors (v-NAR), and bis-single chain Fv regions (see, e.g., Hollinger and Hudson, Nature Biotechnology, 23(9):1126-1136, 2005).
  • antibodies or antigen-binding fragments thereof comprise a light chain and/or a heavy chain constant region, such as one or more constant regions, including one or more IgG1, IgG2, IgG3 and/or IgG4 constant regions.
  • antibodies can include epitope-binding fragments of any of the above.
  • the antibodies described herein can be of any class (e.g., IgG, IgE, IgM, IgD, and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) of immunoglobulin molecule.
  • Antibodies can be antagonistic antibodies or agonistic antibodies.
  • the term “monospecific antibody” refers to an antibody that has one or more binding sites each of which bind to the same epitope of the same antigen, e.g., GPC3 (e.g., human GPC3).
  • the term “multispecific molecule” refers to a molecule (e.g., an antibody) that is capable of binding to at least two distinct antigenic determinants, for example two binding sites each formed by a pair of an antibody heavy chain variable domain (V H ) and an antibody light chain variable domain (V L ) binding to different antigens or to different epitopes on the same antigen. Such a multispecific molecule may have a 1+1 format.
  • Other multispecific molecule formats may be 2+1 or 1+2 formats (comprising two binding sites for a first antigen or epitope and one binding site for a second antigen or epitope) or 2+2 formats (comprising two binding sites for a first antigen or epitope and two binding sites for a second antigen or epitope).
  • a multispecific molecule comprises two antigen binding sites, each may bind to a different antigenic determinant.
  • Such a multispecific molecule may bind to two different epitopes on the same antigen (e.g., epitopes on GPC3) or on different antigens (e.g., an epitope on GPC3 and an epitope on an antigen different from GPC3).
  • epitope refers to the region of an antigen to which an antibody binds preferentially and specifically.
  • a monoclonal antibody binds preferentially to a single specific epitope of a molecule that can be molecularly defined.
  • multiple epitopes can be recognized by a multispecific antibody.
  • antibody fragment refers to any derivative of an antibody which is less than full-length. In exemplary embodiments, the antibody fragment retains at least a significant portion of the full-length antibody’s specific binding ability.
  • antibody fragments include, but are not limited to, Fab, Fab′, F(ab′)2, scFv, Fv, dsFv diabody, Fc, and Fd fragments.
  • the antibody fragment may be produced by any means.
  • the antibody fragment may be enzymatically or chemically produced by fragmentation of an intact antibody, it may be recombinantly produced from a gene encoding the partial antibody sequence, or it may be wholly or partially synthetically produced.
  • the antibody fragment may optionally be a single chain antibody fragment. Alternatively, the fragment may comprise multiple chains which are linked together, for instance, by disulfide linkages.
  • the fragment may also optionally be a multimolecular complex.
  • a functional antibody fragment will typically comprise at least about 50 amino acids and more typically will comprise at least about 200 amino acids.
  • Fab fragment refers to a fragment of an antibody comprising an antigen-binding site generated by cleavage of the antibody with the enzyme papain, which cuts at the hinge region N-terminally to the inter-H-chain disulfide bond and generates two Fab fragments from one antibody molecule.
  • F(ab′)2 fragment refers to a fragment of an antibody containing two antigen-binding sites, generated by cleavage of the antibody molecule with the enzyme pepsin which cuts at the hinge region C-terminally to the inter-H-chain disulfide bond.
  • the term “Fc fragment” refers to the fragment of an antibody comprising the constant domain of its heavy chain.
  • the term “Fv fragment” refers to the fragment of an antibody comprising the variable domains of its heavy chain and light chain.
  • the term “single chain variable fragment” or “scFv” refers to an Fv fragment in which the heavy chain domain and the light chain domain are linked.
  • One or more scFv fragments may be linked to other antibody fragments (such as the constant domain of a heavy chain or a light chain) to form antibody constructs having one or more antigen recognition sites.
  • the heavy chain domain and light chain domain are either joined directly or joined by a linker, which connects the N-terminus of the heavy chain domain with the C-terminus of the light chain domain, or the C-terminus of the heavy chain domain with the N-terminus of the light chain domain.
  • linker is art-recognized and refers to a molecule or group of molecules connecting two NAI-1540294631v3 36 compounds, such as two polypeptides.
  • the linker may be comprised of a single linking molecule or may comprise a linking molecule and a spacer molecule, intended to separate the linking molecule and a compound by a specific distance.
  • the linker is usually rich in glycine for flexibility, as well as serine or threonine for solubility.
  • the linker is a G4S linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, which is provided below: GGGGSGGGGSGGGGS [SEQ ID NO: 1]
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 2, which is provided below: GGGGSGGGGSGGGSGGGGS [SEQ ID NO: 2]
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 3, which is provided below: GGGGSGGGGSGGGGSGGGSGGGGS [SEQ ID NO: 3]
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 4, which is provided below: GGGGSGGGGSGGGGSGGGGSGGGSGGGSG
  • CDRs can be identified according to a number of known numbering systems.
  • the CDRs are identified according to the Kabat numbering system.
  • the Kabat CDRs are based on sequence variability and are the most commonly used (see e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)).
  • the CDRs can be identified according to the Chothia numbering system. Chothia refers to the location of the structural loops (see e.g., Chothia and Lesk, J. Mol. Biol.196:901-917 (1987)).
  • the CDRs can be identified according to the AbM numbering system.
  • the AbM hypervariable regions represent a compromise between the Kabat CDRs and Chothia structural loops, and are used by Oxford Molecular’s AbM antibody modeling software (see e.g., Martin, in Antibody Engineering, Vol.2, Chapter 3, Springer Verlag).
  • the CDRs can be identified according to the Contact numbering system (see, e.g., MacCallum RM et al., 1996, J Mol Biol 5: 732-745).
  • the Contact CDRs are based on an analysis of the available complex crystal structures.
  • the CDRs can be identified according to the ImMunoGeneTics (IMGT) ® Information System. A universal numbering system has been developed and widely adopted, IMGT ® Information System (Lefranc et al., Dev. Comp.
  • IMGT ® is an integrated information system specializing in immunoglobulins (IG), T cell receptors (TR) and major histocompatibility complex (MHC) of human and other vertebrates.
  • IG immunoglobulins
  • TR T cell receptors
  • MHC major histocompatibility complex
  • amino acid abbreviations used herein are conventional one letter codes for the amino acids and are expressed as follows: A, alanine; B, asparagine or aspartic acid; C, cysteine; D aspartic acid; E, glutamate, glutamic acid; F, phenylalanine; G, glycine; H histidine; I isoleucine; K, lysine; L, leucine; M, methionine; N, asparagine; P, proline; Q, glutamine; R, arginine; S, serine; T, threonine; V, valine; W, tryptophan; Y, tyrosine; Z, glutamine or glutamic acid.
  • polypeptide refers to a natural or synthetic molecule comprising two or more amino acids linked by the carboxyl group of one amino acid to the alpha amino group of another.
  • polypeptide fragment refers to a polypeptide in which amino acid residues are deleted as compared to the reference polypeptide itself, but where the remaining amino acid sequence is usually identical to that of the reference polypeptide. Such deletions may occur at the amino-terminus or carboxy-terminus of the reference polypeptide, or alternatively both.
  • Fragments typically are at least about 5, 6, 8 or 10 amino acids long, at least about 14 amino acids long, at least about 20, 30, 40 or 50 amino acids long, at least about 75 amino acids long, or at least about 100, 150, 200, 300, 500 or more amino acids long.
  • a fragment can retain one or more of the biological activities of the reference polypeptide.
  • a fragment may comprise an enzymatic activity and/or an interaction site of the reference polypeptide.
  • a fragment may have immunogenic properties.
  • the term “substantially identical” or “substantially homologous” refers to a polypeptide or a nucleic acid molecule exhibiting at least about 50% identical or homologous to a reference amino acid sequence (for example, any of the amino acid sequences described herein) or a reference nucleotide sequence (for example, any of the nucleic acid sequences described herein). In certain embodiments, such a sequence is at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, or at least about 100% identical or homologous to the amino acid sequence or the nucleic acid sequence used for comparison.
  • identity refers to sequence identity between two nucleic acid molecules or polypeptides. Identity can be determined by comparing a position in each sequence which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base, then the molecules are identical at that position. A degree of similarity or identity between nucleic acid or amino acid sequences is a function of the number of identical or matching nucleotides at positions shared by the nucleic acid sequences.
  • Various alignment algorithms and/or programs may be used to calculate the identity between two sequences, including FASTA, or BLAST which are available as a part of the GCG sequence analysis package (University of Wisconsin, Madison, Wis.), and can be used with, e.g., default setting.
  • BLASTP the percent similarity is based on the BLASTP positives score and the percent sequence identity is based on the BLASTP identities score.
  • BLASTP “Identities” shows the number and fraction of total NAI-1540294631v3 39 residues in the high scoring sequence pairs which are identical; and BLASTP “Positives” shows the number and fraction of residues for which the alignment scores have positive values and which are similar to each other.
  • nucleic acid refers to a natural or synthetic molecule comprising a single nucleotide or two or more nucleotides linked by a phosphate group at the 3’ position of one nucleotide to the 5’ end of another nucleotide.
  • the nucleic acid is not limited by length, and thus the nucleic acid can include deoxyribonucleic acid (DNA) or ribonucleic acid (RNA).
  • the term “operably linked to” refers to the functional relationship of a nucleic acid with another nucleic acid sequence. Promoters, enhancers, transcriptional and translational stop sites, and other signal sequences are examples of nucleic acid sequences operably linked to other sequences.
  • operable linkage of DNA to a transcriptional control element refers to the physical and functional relationship between the DNA and promoter such that the transcription of such DNA is initiated from the promoter by an RNA polymerase that specifically recognizes, binds to and transcribes the DNA.
  • signal sequence or “leader sequence” refers to a peptide sequence (e.g., 5, 10, 15, 20, 25 or 30 amino acids) present at the N-terminus of proteins that directs their entry to the secretory pathway.
  • variant refers to an amino acid or peptide sequence having conservative amino acid substitutions, non-conservative amino acid substitutions (i.e. a degenerate variant), substitutions within the wobble position of each codon (i.e.
  • DNA and RNA encoding an amino acid, amino acids added to the C-terminus of a peptide, or a peptide having 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% sequence identity to a reference sequence.
  • a conservative sequence modification refers to an amino acid modification that does not significantly affect or alter the binding characteristics of the presently disclosed GPC3-targeted binding comprising the amino acid sequence.
  • Conservative modifications can include amino acid substitutions, additions and deletions. Modifications can be introduced by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis.
  • Amino acids can be classified into groups according to their physicochemical properties such as charge and polarity. Conservative amino acid substitutions are ones in which the NAI-1540294631v3 40 amino acid residue is replaced with an amino acid within the same group.
  • amino acids can be classified by charge: positively-charged amino acids include lysine, arginine, histidine, negatively-charged amino acids include aspartic acid, glutamic acid, neutral charge amino acids include alanine, asparagine, cysteine, glutamine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.
  • amino acids can be classified by polarity: polar amino acids include arginine (basic polar), asparagine, aspartic acid (acidic polar), glutamic acid (acidic polar), glutamine, histidine (basic polar), lysine (basic polar), serine, threonine, and tyrosine; non-polar amino acids include alanine, cysteine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, and valine.
  • one or more amino acid residues within a CDR region can be replaced with other amino acid residues from the same group and the altered antibody can be tested for retained function using the functional assays described herein.
  • no more than one, no more than two, no more than three, no more than four, no more than five residues within a specified sequence or a CDR region are altered.
  • vector refers to a nucleic acid sequence capable of transporting into a cell another nucleic acid to which the vector sequence has been linked.
  • expression vector includes any vector, (e.g., a plasmid, cosmid or phage chromosome) containing a gene construct in a form suitable for expression by a cell (e.g., linked to a transcriptional control element).
  • an effective amount refers to an amount sufficient to affect a beneficial or desired clinical result upon treatment.
  • An effective amount can be administered to a subject in one or more doses.
  • an effective amount can be an amount that is sufficient to palliate, ameliorate, stabilize, reverse or slow the progression of the disease, or otherwise reduce the pathological consequences of the disease.
  • the effective amount can be determined by a physician on a case-by-case basis and is within the skill of one in the art. Several factors are typically taken into account when determining an appropriate dosage to achieve an effective amount. These factors include age, sex and weight of the subject, the condition being treated, the severity of the condition and the form and effective concentration of the cells administered. [00118]
  • pharmaceutically acceptable refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit/risk ratio.
  • treatment refers to the medical management of a subject with the intent to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder.
  • This term includes active treatment, that is, treatment directed specifically toward the improvement of a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder.
  • this term includes palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder.
  • An “individual” or “subject” herein is a vertebrate, such as a human or a non-human animal, for example, a mammal. Mammals include, but are not limited to, humans, primates, farm animals, sport animals, rodents and pets.
  • Non-limiting examples of non-human animal subjects include rodents such as mice, rats, hamsters, and guinea pigs, rabbits, dogs, cats, sheep, pigs, goats, cattle, horses, and non-human primates such as apes and monkeys.
  • the subject is a mammal.
  • the subject is a human.
  • the term “GPC3 antigen” and, in particular, “GPC3” is intended to include fragments, variants (e.g., allelic variants), and derivatives of the antigen molecule, e.g., the GPC3 molecule.
  • GPC3 is also known as glypican-3.
  • Glypican-3 Glypican-3 (GPC3) is a 70 kDa protein that is anchored to the cell surface via glycosylphosphatidylinositol (GPI). GPC3 is widely expressed in the placenta and fetal tissues such as liver, lung, and kidney, and it regulates morphogenesis or growth. However, its expression is significantly reduced in adult organs.
  • GPC3 is a member of the heparan sulfate (HS) proteoglycan family that consists of six members, GPC1 to GPC6, which serve as receptors or coreceptors for a number of ligand molecules, including morphogens, growth factors, adhesion, and matrix molecules, to regulate their signaling and distribution.
  • HS heparan sulfate
  • the presently disclosed binding molecule binds to a human GPC3 antigen.
  • the GPC3 is a human GPC3.
  • the GPC3 is a wild-type human GPC3.
  • the GPC3 antigen comprises or consists of the amino acid sequence with a Uniprot Reference No: P51654-1 (SEQ ID NO: 22), which is provided below.
  • the presently disclosed binding molecule binds to an epitope of GPC3.
  • the epitope is within amino acids 524 to 563 of SEQ ID NO: 22.
  • the epitope is at the C-terminus of GPC3.
  • Chimeric Antigen Receptors [00127] The presently disclosed subject matter provides chimeric antigen receptors (CARs) that targets GPC3. [00128] CARs are engineered receptors, which graft or confer a specificity of interest onto an immune effector cell.
  • the GPC3-targeted CAR comprises an extracellular domain that specifically binds to GPC3, a transmembrane domain, and an intracellular domain.
  • the extracellular domain is fused to the transmembrane domain, which is fused to the intracellular domain.
  • the extracellular domain of the CAR comprises a GPC3-binding region, and is responsible for antigen recognition.
  • the extracellular domain further comprise a signal peptide (SP) so that the CAR can be glycosylated and anchored in the cell membrane of a cell (e.g., an immunoresponsive cell).
  • SP signal peptide
  • the transmembrane domain (TD) connects the extracellular domain to the intracellular domain and resides within the cell membrane when expressed by a cell (e.g., an immunoresponsive cell).
  • the intracellular domain transmits an activation signal to the cell (e.g., an immunoresponsive cell) after antigen recognition.
  • the intracellular domain comprises an intracellular signaling domain (ISD). In certain embodiments, the intracellular domain further comprises a co-stimulatory signaling region (CSR).
  • a “signaling domain (SD)” generally comprises immunoreceptor tyrosine-based activation motifs (ITAMs) that activate a signaling cascade when the ITAM is phosphorylated.
  • the term “co-stimulatory signaling region (CSR)” refers to an intracellular signaling domain from a costimulatory protein receptor, such as CD28, 41BB, and ICOS, that are able to enhance the activation of an immunoresponsive cell (e.g., a T cell).
  • the CAR can be for example (and without limitation), a TRUCK, Universal CAR, a Self-driving CAR, an Armored CAR, a Self-destruct CAR, a Conditional CAR, a Marked CAR, a TenCAR, a Dual CAR, or a sCAR.
  • TRUCKs T cells redirected for universal cytokine killing
  • CAR chimeric antigen receptor
  • Cytokine expression may be constitutive or induced by T cell activation. Targeted by CAR specificity, localized production of pro-inflammatory cytokines recruits endogenous immune cells to tumor sites and may potentiate an antitumor response.
  • Universal, allogeneic CAR T cells are engineered to no longer express endogenous T cell receptor (TCR) and/or major histocompatibility complex (MHC) molecules, thereby preventing graft-versus-host disease (GVHD) or rejection, respectively.
  • Self-driving CARs co-express a CAR and a chemokine receptor, which binds to a tumor ligand, thereby enhancing tumor homing.
  • CAR T cells engineered to be resistant to immunosuppression may be genetically modified to no longer express various immune checkpoint molecules (e.g., cytotoxic T lymphocyte-associated antigen 4 (CTLA4) or programmed cell death protein 1 (PD-1)).
  • immune checkpoint molecules e.g., cytotoxic T lymphocyte-associated antigen 4 (CTLA4) or programmed cell death protein 1 (PD-1)
  • CTL4 cytotoxic T lymphocyte-associated antigen 4
  • PD-1 programmed cell death protein 1
  • Exemplary “Knockdown” and “Knockout” techniques include, but are not limited to, RNA interference (RNAi) (e.g., asRNA, miRNA, shRNA, siRNA, etc.) and CRISPR interference (CRISPRi) (e.g., CRISPR- Cas9).
  • CRISPRi CRISPR interference
  • CRISPRi CRISPR interference- Cas9
  • the extracellular ligand-binding domain of the immune checkpoint molecule is fused to a transmembrane membrane in order to compete for ligand binding.
  • the extracellular ligand-binding domain of PD-1 may be fused to a CD8 transmembrane domain, thus competing for PD-1 ligand from the target cell.
  • CAR T cells are engineered to express an immune checkpoint switch receptor to exploit the inhibitory immune checkpoint ligand present on a target cell.
  • the extracellular ligand-binding domain of the immune checkpoint molecule is fused to a signaling, stimulatory, and/or co-stimulatory domain.
  • the extracellular ligand-binding domain of PD-1 may be fused to a CD28 domain, thus providing CD28 co-stimulation while blocking PD-1 signaling.
  • the CAR T cells may be administered with an aptamer or a monoclonal antibody that blocks immune checkpoint signaling.
  • the CAR T cells e.g., CAR T cell therapy
  • the CAR T cells and PD-1 pathway-blocking antibodies are administered conjointly.
  • inducible apoptosis of the T cell may be achieved based on ganciclovir binding to thymidine kinase in gene-modified lymphocytes or the more recently described system of activation of human caspase 9 by a small-molecule dimerizer.
  • a conditional CAR T cell is by default unresponsive, or switched “off”, until the addition of a small molecule to complete the circuit, enabling full transduction of both signal 1 and signal 2, thereby activating the CAR T cell.
  • T cells may be engineered to express an adaptor- specific receptor with affinity for subsequently administered secondary antibodies directed at target antigen.
  • Marked CAR T cells express a CAR plus a tumor epitope to which an existing monoclonal antibody agent binds. In the setting of intolerable adverse effects, administration of the monoclonal antibody clears the CAR T cells and alleviates symptoms with no additional off-tumor effects.
  • a tandem CAR (TanCAR) T cell expresses a single CAR consisting of two linked single- chain variable fragments (scFvs) that have different affinities fused to intracellular co-stimulatory domain(s) and a CD3 ⁇ domain. TanCAR T cell activation is achieved only when target cells co- express both targets.
  • a dual CAR T cell expresses two separate CARs with different ligand binding targets; one CAR includes only the CD3 ⁇ domain and the other CAR includes only the co-stimulatory domain(s). Dual CAR T cell activation requires co-expression of both targets on the tumor.
  • a safety CAR consists of an extracellular scFv fused to an intracellular inhibitory domain. sCAR T cells co-expressing a standard CAR become activated only when encountering target cells that possess the standard CAR target but lack the sCAR target.
  • the CAR comprises one signaling domain.
  • the CAR comprises one or more signaling domain (co-stimulatory signaling region).
  • the one or more signaling domain may be a polypeptide selected from the group consisting of CD8, CD3 ⁇ , CD3 ⁇ , CD3 ⁇ , CD3 ⁇ , Fc ⁇ RI- ⁇ , FcyRIII-y, Fc ⁇ RI ⁇ , Fc ⁇ RIy, DAP10, DAP12, CD32, CD79a, CD79b, CD28, CD3C, CD4, B2C, 4-1BB (CD137), ICOS, CD27, CD288, CD80, NKp30, OX40, and mutants thereof. 5.3.1.
  • the extracellular domain of the CARs disclosed herein generally comprise an antigen recognition domain that binds a GPC3 antigen. Such antigen-specific binding domains are typically derived from an antibody.
  • the extracellular domain of the CAR comprises or is a functional antibody fragment or derivative thereof (e.g., a single chain variable fragment (scFv) or a Fab, or any suitable antigen binding fragment of an antibody).
  • the extracellular domain of the CAR comprises an scFv.
  • the scFv is from a monoclonal antibody (mAb).
  • the scFv can be a human scFv, a humanized scFv, or a murine scFv.
  • the scFv is a human scFv.
  • the scFv can be derived from fusing the heavy chain variable region (V H ) and light chain variable region (V L ) of an antibody.
  • the scFv may be derived from Fab’s (instead of from an antibody, e.g., obtained from Fab libraries).
  • the extracellular domain of the CAR comprises a Fab.
  • the Fab is crosslinked.
  • the extracellular domain of the CAR comprises a F(ab)2.
  • Any of the foregoing molecules may be comprised in a fusion protein with a heterologous sequence to form the extracellular domain of the CAR.
  • the extracellular domain of the CAR binds to a GPC3 antigen (e.g., a human GPC3 antigen) with a EC50 of between about 1 ng/mL and about 600 ng/mL, between about 1 ng/mL and about 500 ng/mL, between about 1 ng/mL and about 400 ng/mL, between about 1 ng/mL and about 300 ng/mL, between about 1 ng/mL and about 200 ng/mL, between about 1 ng/mL and about 150 ng/mL between about 1 ng/mL and about 100 ng/mL, between about 1 ng/mL and about 60 ng/mL, between about 1 ng/mL and
  • a GPC3 antigen e.g
  • the extracellular domain of the CAR binds to GPC3 antigen (e.g., a human GPC3 antigen) with a EC50 of between about 1 ng/mL and about 60 ng/mL. In certain embodiments, the extracellular domain of the CAR binds to GPC3 antigen (e.g., a human GPC3 antigen) with a EC50 of between about 4 ng/mL and about 40 ng/mL.
  • GPC3 antigen e.g., a human GPC3 antigen
  • Binding of the extracellular domain of the CAR can be confirmed by, for example, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), FACS analysis, bioassay (e.g., growth inhibition), or Western Blot assay.
  • ELISA enzyme-linked immunosorbent assay
  • RIA radioimmunoassay
  • FACS analysis bioassay (e.g., growth inhibition)
  • bioassay e.g., growth inhibition
  • Western Blot assay Western Blot assay.
  • Each of these assays generally detect the presence of protein-antibody complexes of particular interest by employing a labeled reagent (e.g., an antibody, or a scFv) specific for the complex of interest.
  • a labeled reagent e.g., an antibody, or a scFv
  • the scFv can be radioactively labeled and used in a radioimmunoassay (RIA) (see, for example, Weintraub, B., Principles of Radioimmunoassays, Seventh Training Course on Radioligand Assay Techniques, The Endocrine Society, March, 1986, which is incorporated by reference herein).
  • the radioactive isotope can be detected by such means as the use of a ⁇ counter or a scintillation counter or by autoradiography.
  • the GPC3-targeted extracellular domain is labeled with a fluorescent marker.
  • fluorescent markers include green fluorescent protein (GFP), blue fluorescent protein (e.g., EBFP, EBFP2, Azurite, and mKalama1), cyan fluorescent protein (e.g., ECFP, Cerulean, and CyPet), and yellow fluorescent protein (e.g., YFP, Citrine, Venus, and YPet).
  • GFP green fluorescent protein
  • blue fluorescent protein e.g., EBFP, EBFP2, Azurite, and mKalama1
  • cyan fluorescent protein e.g., ECFP, Cerulean, and CyPet
  • yellow fluorescent protein e.g., YFP, Citrine, Venus, and YPet.
  • V H heavy chain variable region
  • V L light chain variable region
  • the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a V H comprising the amino acid sequence of SEQ ID NO: 29; and/or ii) the V L comprises a V L CDR1, a V L CDR2, and a V L CDR3 as set forth in a V L comprising the amino acid sequence of SEQ ID NO: 30.
  • SEQ ID NO: 29 and SEQ ID NO: 30 are disclosed in Table 1.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25.
  • SEQ ID NOs: 23-25 are provided in Table 1.
  • the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29.
  • the V H comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 29.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28.
  • V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30.
  • the V L comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30.
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 30.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 NAI-1540294631v3 49 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 29
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 30.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the VH and VL are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be V H -V L or V L -V H .
  • the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28.
  • the scFv comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 29, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 30.
  • the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31.
  • SEQ ID NO: 31 is provided in Table 1.
  • the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- V H .
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28.
  • the scFv comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 29, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 30.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 32.
  • SEQ ID NO: 32 is provided in Table 1. [00167]
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 29 an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 29 is set forth in SEQ ID NO: 33, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 29 is set forth in SEQ ID NO: 34, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 30 is set forth in SEQ ID NO: 35, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 30 is set forth in SEQ ID NO: 36, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 31 is set forth in SEQ ID NO: 37, which is provided below.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43.
  • the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 47.
  • the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 47.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 47.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, NAI-1540294631v3 54 and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • SEQ ID NOs: 44-46 are provided in Table 2.
  • the V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 48.
  • the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 48.
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 48.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 47
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 48.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the VH and VL are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be V H -V L or V L -V H .
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the NAI-1540294631v3 55 amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 47, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 48.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 49.
  • SEQ ID NO: 49 is provided in Table 2.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 47, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 48.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 50.
  • SEQ ID NO: 50 is provided in Table 2. [00186] In certain embodiments, the CDRs are identified according to the Kabat numbering system.
  • Table 2 (ATA002 or ATA018) CDRs 1 2 3 VI TYN NTNNN KFK TTY E ID NAI-1540294631v3 56 V H -V L QVQLQQSGPEVVRPGVSVKISCKGSGYTFTDYAIHWVKQSHAKSLEWIGVISTY scFv NGNTNNNQKFKGKATMTVDKSSSTAYLELARLTSEDSAIYYCARSGTTYWGQGT A i TLTVSSGGGGSGGGGSGGGGSDVVMTQTPLTLSVTIGQPASISCKSSQSLLDSE , sequence of SEQ ID NO: 47 is set forth in SEQ ID NO: 51, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 47 is set forth in SEQ ID NO: 52, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 48 is set forth in SEQ ID NO: 53, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 48 is set forth in SEQ ID NO: 54, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 49 is set forth in SEQ ID NO: 55, which is provided below.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61.
  • VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 62.
  • the V H comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 62.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 62.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • SEQ ID NOs: 26, 27, and 46 are provided in Table 3.
  • the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 63.
  • the V L comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous NAI-1540294631v3 60 to the amino acid sequence set forth in SEQ ID NO: 63.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 63.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof; and V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 62
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 63.
  • the VH the VL can be linked one after another, e.g., by a linker.
  • the V H and V L are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be VH-VL or VL-VH.
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the V H and the V L are positioned from the N- to the C-terminus as V H -V L .
  • the scFv comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 62, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 63.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid NAI-1540294631v3 61 sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 64.
  • SEQ ID NO: 64 is provided in Table 3.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 62, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 63.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 65.
  • SEQ ID NO: 65 is provided in Table 3. [00205] In certain embodiments, the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 62 CDRs 1 2 3 V H DYAVH (SEQ ID NO: VISTYNGNTNYNQKFKG SGTSY (SEQ ID Q NAI-1540294631v3 62 [00206]
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 62 is set forth in SEQ ID NO: 66, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 62 is set forth in SEQ ID NO: 67, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 63 is set forth in SEQ ID NO: 68, which is provided below.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, NAI-1540294631v3 65 and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25.
  • SEQ ID NOs: 23, 60, and 25 are provided in Table 4.
  • the V H comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 75.
  • the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 75.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 75.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • SEQ ID NOs: 74, 45, and 46 are provided in Table 4.
  • the V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 76.
  • the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 76.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 76.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification, and a CDR3 comprising the amino acid sequence set NAI-1540294631v3 66 forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 75
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 76.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the V H and V L are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be VH-VL or VL-VH.
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 76.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 77. SEQ ID NO: 77 is provided in Table 4.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 76.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 78.
  • SEQ ID NO: 78 is provided in Table 4. [00224]
  • the CDRs are identified according to the Kabat numbering system.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 75 is set forth in SEQ ID NO: 79, which is provided below CAGGTCCAACTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTATGCTATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATCAGTACTTACAATGGTAATACAAACTACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAGG ATTCTGCCATCTATTACTGTCTAAAGAACGACTACTGGGGTCAAG
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 76 is set forth in SEQ ID NO: 82, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 77 is set forth in SEQ ID NO: 83, which is provided below.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43.
  • V H comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 89.
  • the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 89.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 89.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set NAI-1540294631v3 71 forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 90.
  • the V L comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 90.
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 90.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 89
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 90.
  • the VH the VL can be linked one after another, e.g., by a linker.
  • the VH and VL are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be V H -V L or V L -V H .
  • the VH is positioned at the N-terminus of the extracellular domain, i.e., the V H and the V L are positioned from the N- to the C-terminus as V H -V L .
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a NAI-1540294631v3 72 CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 89, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 90.
  • the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 91. SEQ ID NO: 91 is provided in Table 5.
  • the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- V H .
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 89, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 90.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 92.
  • SEQ ID NO: 92 is provided in Table 5. [00243] In certain embodiments, the CDRs are identified according to the Kabat numbering system.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 89 is set forth in SEQ ID NO: 94, which is provided below CAGGTGCAGCTGCAGCAGTCCGGCCCCGAGGTGGAAAGACCTGGCGTTAGCGTGAAGATCA
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 90 is set forth in SEQ ID NO: 96, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 91 is set forth in SEQ ID NO: 97, which is provided below.
  • the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 101.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • SEQ ID NOs: 44-46 are provided in Table 6.
  • the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 102.
  • the V L comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or NAI-1540294631v3 77 homologous to the amino acid sequence set forth in SEQ ID NO: 102.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 102.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 101
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 102.
  • the VH the VL can be linked one after another, e.g., by a linker.
  • the V H and V L are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be VH-VL or VL-VH.
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the V H and the V L are positioned from the N- to the C-terminus as V H -V L .
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 101, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 102.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid NAI-1540294631v3 78 sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 103.
  • SEQ ID NO: 103 is provided in Table 6.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 101, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 102.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 104.
  • SEQ ID NO: 104 is provided in Table 6. [00262]
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 101 an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 101 is set forth in SEQ ID NO: 105, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 101 is set forth in SEQ ID NO: 106, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 102 is set forth in SEQ ID NO: 107, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 102 is set forth in SEQ ID NO: 108, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 103 is set forth in SEQ ID NO: 109, which is provided below.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: NAI-1540294631v3 82 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115.
  • SEQ ID NOs: 113-115 are provided in Table 7.
  • the V H comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 119.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • SEQ ID NOs: 116-118 are provided in Table 7.
  • the V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 120.
  • the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 120.
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 120.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification, and a CDR3 comprising the amino acid sequence NAI-1540294631v3 83 set forth in SEQ ID NO: 118 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 119
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 120.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the VH and VL are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be V H -V L or V L -V H .
  • the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the scFv comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 119, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 120.
  • the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 121.
  • SEQ ID NO: 121 is provided in Table 7.
  • the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- V H .
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set NAI-1540294631v3 84 forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 120.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 122.
  • SEQ ID NO: 122 is provided in Table 7. [00281]
  • the CDRs are identified according to the Kabat numbering system.
  • Table 7 (ATA007) CDRs 1 2 3 V H TYGVH (SEQ ID NO: VIWSGGSTDYNASFIS RGNDGPPFAY Q G Q D G Q Y Y D G K R [00282]
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 119 is set forth in SEQ ID NO: 123, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 119 is set forth in SEQ ID NO: 124, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 120 is set forth in SEQ ID NO: 125, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 120 is set forth in SEQ ID NO: 126, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 121 is set forth in SEQ ID NO: 127, which is provided below.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133.
  • SEQ ID NOs: 131-133 are provided in Table 8.
  • the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 137.
  • the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 137.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 137.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino NAI-1540294631v3 88 acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
  • V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 138.
  • the V L comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 138.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 138.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof; and V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 137
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 138.
  • the VH the VL can be linked one after another, e.g., by a linker.
  • the V H and V L are linked via a linker.
  • the linker comprises or consists of NAI-1540294631v3 89 the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be VH-VL or VL-VH.
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 138.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 139.
  • SEQ ID NO: 139 is provided in Table 8.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the V H and the V L are positioned from the N- to the C-terminus as V L - VH.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 138.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 140.
  • SEQ ID NO: 140 is provided in Table 8. [00300] In certain embodiments, the CDRs are identified according to the Kabat numbering system.
  • Table 8 (ATA008 or ATA020) CDRs 1 2 3 D N - 5 0 9 63 v3 90 VL RASGNIHNYLA (SEQ NAKTLAD (SEQ ID NO: QHFWTTPFT (SEQ ID NO: 134) 135) ID NO: 136) V LK SGPGLV PS SLSITCTVSGFSLNSYGVHWVR SPGKGLEWLGAIWSSG M A G M G D A G K N sequence of SEQ ID NO: 137 is set forth in SEQ ID NO: 141, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 137 is set forth in SEQ ID NO: 142, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 138 is set forth in SEQ ID NO: 143, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 138 is set forth in SEQ ID NO: 144, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 139 is set forth in SEQ ID NO: 145, which is provided below.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151.
  • the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 155.
  • the V H comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 155.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 155.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154.
  • SEQ ID NOs: 152-154 are provided in Table 9.
  • the V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 156.
  • the VL comprises an amino NAI-1540294631v3 94 acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 156.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 156.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof; and V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 155
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 156.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the V H and V L are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be VH-VL or VL-VH.
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 156.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 157. SEQ ID NO: 157 is provided in Table 9.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the V H and the V L are positioned from the N- to the C-terminus as V L - VH.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 156.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 158.
  • SEQ ID NO: 158 is provided in Table 9. [00319] In certain embodiments, the CDRs are identified according to the Kabat numbering system.
  • Table 9 (ATA009 or ATA021) CDRs 1 2 3 V DAYIH (SEQ ID NO: GIDPENDNTEYDPKFPG RPLGLPFDY : : N Q F L N Q S E NAI-1540294631v3 96 V L -V H DIVLSQSPSSLAVSVGENVTMNCKSSQRLLDSSNQRNYLAWYQQKPGQSPQLLIF scFv WASTRESGVPDRFTGSGSGTDFTLTIRSVKAEDLAVYYCQQYYSYPFTFGSGTKL A i EIKGGGGSGGGGSGGGGSEVQLQQSGAEHVKPGTSVRLSCTASGFNIKDAYIHWV A sequence of SEQ ID NO: 155 is set forth in SEQ ID NO: 159, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 155 is set forth in SEQ ID NO: 160, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 156 is set forth in SEQ ID NO: 161, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 156 is set forth in SEQ ID NO: 162, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 157 is set forth in SEQ ID NO: 163, which is provided below.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 NAI-1540294631v3 99 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168.
  • SEQ ID NOs: 131, 167, and 168 are provided in Table 10.
  • the V H comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 170.
  • the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 170.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 170.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • SEQ ID NOs: 134, 135, and 169 are provided in Table 10.
  • the V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 171.
  • the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 171.
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 171.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification NAI-1540294631v3 100 thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 170
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 171.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the VH and VL are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be V H -V L or V L -V H .
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 170, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 171.
  • the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 172.
  • SEQ ID NO: 172 is provided in Table 10.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- V H .
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 NAI-1540294631v3 101 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 170, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 171.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 173.
  • SEQ ID NO: 173 is provided in Table 10. [00338]
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 170 an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 170 is set forth in SEQ ID NO: 174, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 170 is set forth in SEQ ID NO: 175, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 171 is set forth in SEQ ID NO: 176, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 171 is set forth in SEQ ID NO: 177, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 172 is set forth in SEQ ID NO: 178, which is provided below.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182.
  • V H comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 183.
  • the V H comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about NAI-1540294631v3 105 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 183.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 183.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • SEQ ID NOs: 26, 27, and 46 are provided in Table 11.
  • the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 184.
  • the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 184.
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 184.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 183
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 184.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the VH and VL are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be V H -V L or V L -V H .
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 184.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 185.
  • SEQ ID NO: 185 is provided in Table 11.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- V H .
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 184.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 186.
  • SEQ ID NO: 186 is provided in Table 11. [00357]
  • the CDRs are identified according to the Kabat numbering system.
  • Table 11 (ATA011) CDRs 1 2 3 V H DYAIH (SEQ ID NO: VISTYNGNTNYNRKFKD DDY (SEQ ID Q N V L E N V L C L E K I [00358]
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 183 is set forth in SEQ ID NO: 187, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 183 is set forth in SEQ ID NO: 188, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 184 is set forth in SEQ ID NO: 189, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 184 is set forth in SEQ ID NO: 190, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 185 is set forth in SEQ ID NO: 191, which is provided below.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197.
  • SEQ ID NOs: 195-197 are provided in Table 12.
  • the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 201.
  • the V H comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 201.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 201.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200.
  • SEQ ID NOs: 198-200 are provided in Table 12.
  • the V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 202.
  • the VL comprises an amino NAI-1540294631v3 111 acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 202.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 202.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof; and V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 201
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 202.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the V H and V L are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be VH-VL or VL-VH.
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 202.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 203.
  • SEQ ID NO: 203 is provided in Table 12.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the V H and the V L are positioned from the N- to the C-terminus as V L - VH.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 202.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 204.
  • SEQ ID NO: 204 is provided in Table 12. [00376]
  • the CDRs are identified according to the Kabat numbering system.
  • Table 12 (ATA012) CDRs 1 2 3 V SYYMS (SEQ ID NO: AINFYGGLAYYPDTVKG HYRYDNAMDY Q G G Y G G T Y Y G 113 Amino GSGGGGSGGGGSDVKLVESGGGLVKLGGSLKLSCAASGFTFSSYYMSWVRQTPEK Acid RLDLVAAINFYGGLAYYPDTVKGRFTISRDNAKNTLYLQMNSLKSEDTALYYCVR HYRYDNAMDYWGQGTSVTVSS (SEQ ID NO 204) sequence of SEQ ID NO: 201 is set forth in SEQ ID NO: 205, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 201 is set forth in SEQ ID NO: 206, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 202 is set forth in SEQ ID NO: 207, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 202 is set forth in SEQ ID NO: 208, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 203 is set forth in SEQ ID NO: 209, which is provided below.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 NAI-1540294631v3 116 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168.
  • SEQ ID NOs: 213, 167, and 168 are provided in Table 13.
  • the V H comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 215.
  • the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 215.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 215.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 216.
  • the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 216.
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 216.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification NAI-1540294631v3 117 thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 215, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 216.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the VH and VL are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be V H -V L or V L -V H .
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 215, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 216.
  • the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 217. SEQ ID NO: 217 is provided in Table 13.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- V H .
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 NAI-1540294631v3 118 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 215, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 216.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 218.
  • SEQ ID NO: 218 is provided in Table 13. [00395]
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 215 an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 215 is set forth in SEQ ID NO: 219, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 215 is set forth in SEQ ID NO: 220, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 216 is set forth in SEQ ID NO: 222, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 217 is set forth in SEQ ID NO: 223, which is provided below.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229.
  • SEQ ID NOs: 227-229 are provided in Table 14.
  • the V H comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 233.
  • the V H comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about NAI-1540294631v3 122 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 233.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 233.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232.
  • SEQ ID NOs: 230-232 are provided in Table 14.
  • the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 234.
  • the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 234.
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 234.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 233
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 234.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the VH and VL are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be V H -V L or V L -V H .
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 233, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 234.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 235.
  • SEQ ID NO: 235 is provided in Table 14.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- V H .
  • the extracellular domain of the CAR is an scFv that comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 233, and a V L comprising the amino acid sequence set forth in SEQ ID NO: 234.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 236.
  • SEQ ID NO: 236 is provided in Table 14. [00414]
  • the CDRs are identified according to the Kabat numbering system.
  • NAI-1540294631v3 124 Table 14 (ATA014) CDRs 1 2 3 V H NYWMH (SEQ ID NO: NIYPGSCTSNYDERFKN GNYDYYFDY (SEQ Q S Q H S Q W F H G Q R [00415]
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 233 is set forth in SEQ ID NO: 237, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 233 is set forth in SEQ ID NO: 238, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 234 is set forth in SEQ ID NO: 239, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 234 is set forth in SEQ ID NO: 240, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 235 is set forth in SEQ ID NO: 241, which is provided below.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115.
  • SEQ ID NOs: 113, 245, and 115 are provided in Table 15.
  • the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 247.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • SEQ ID NOs: 116-118 are provided in Table 15.
  • the V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 247.
  • the VL comprises an amino NAI-1540294631v3 128 acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 247.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 247.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 246, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 247.
  • the V H the V L can be linked one after another, e.g., by a linker.
  • the V H and V L are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be VH-VL or VL-VH.
  • the V H is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 247.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 248.
  • SEQ ID NO: 248 is provided in Table 15.
  • the V L is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the V H and the V L are positioned from the N- to the C-terminus as V L - VH.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 247.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 249.
  • SEQ ID NO: 249 is provided in Table 15. [00433]
  • the CDRs are identified according to the Kabat numbering system.
  • Table 15 (ATA015) CDRs 1 2 3 V TYGVH (SEQ ID NO: VIWSGGSTDYNAAFIS RGNDGPPFAY Q G Q D G Q Y Y D G 130 Amino GSGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTTYGVHWVRQSPGK Acid GLDWLGVIWSGGSTDYNAAFISRLSISKDNSKSQVFFKMNSLQTNDTAIYYCARR GNDGPPFAYWGQGTLVTVSA (SEQ ID NO 249) sequence of SEQ ID NO: 246 is set forth in SEQ ID NO: 250, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 246 is set forth in SEQ ID NO: 251, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 247 is set forth in SEQ ID NO: 252, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 247 is set forth in SEQ ID NO: 253, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 248 is set forth in SEQ ID NO: 254, which is provided below.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260.
  • SEQ ID NOs: 258-260 are provided in Table 16.
  • the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 263.
  • the V H comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 263.
  • the VH comprises the amino acid sequence set forth in SEQ ID NO: 263.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
  • SEQ ID NOs: 230, 231, and 261 are provided in Table 16.
  • the V L comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 263.
  • the V L comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 263.
  • the VL comprises the amino acid sequence set forth in SEQ ID NO: 263.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof; and V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 NAI-1540294631v3 134 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
  • the V H comprises the amino acid sequence set forth in SEQ ID NO: 262
  • the V L comprises the amino acid sequence set forth in SEQ ID NO: 63.
  • the VH the VL can be linked one after another, e.g., by a linker.
  • the V H and V L are linked via a linker.
  • the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4.
  • the variable regions from the N- to the C-terminus can be VH-VL or VL-VH.
  • the VH is positioned at the N-terminus of the extracellular domain, i.e., the V H and the V L are positioned from the N- to the C-terminus as V H -V L .
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
  • the scFv comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 262, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 263.
  • the V H and V L are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 264.
  • SEQ ID NO: 264 is provided in Table 16.
  • the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the V H and the V L are positioned from the N- to the C-terminus as V L - VH.
  • the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
  • the scFv comprises a V H comprising the amino acid sequence set forth NAI-1540294631v3 135 in SEQ ID NO: 262, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 263.
  • the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 265.
  • SEQ ID NO: 265 is provided in Table 16. [00452]
  • the CDRs are identified according to the Kabat numbering system.
  • Table 16 (ATA0016 or ATA022) CDRs 1 2 3 V H NYWLH (SEQ ID NO: NIYPGYGTSNYDEKFTN GGYDYYFDY (SEQ Q Y Q H Y Q W Y H G Q R [00453]
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 262 is set forth in SEQ ID NO: 266, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 262 is set forth in SEQ ID NO: 267, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 263 is set forth in SEQ ID NO: 268, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 263 is set forth in SEQ ID NO: 269, which is provided below.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 264 is set forth in SEQ ID NO: 270, which is provided below.
  • a total of 1 to 10 amino acids are substituted, inserted and/or deleted in a specific sequence (e.g., SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, SEQ ID NO: 262, SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ
  • the extracellular domain comprises V H and/or V L sequence selected from SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NAI-1540294631v3 139 NO: 246, SEQ ID NO: 262, SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138
  • the extracellular domain can comprise a leader or a signal peptide that directs the nascent protein into the endoplasmic reticulum.
  • Signal peptide or leader can be essential if the CAR is to be glycosylated and anchored in the cell membrane.
  • the signal sequence or leader can be a peptide sequence (about 5, about 10, about 15, about 20, about 25, or about 30 amino acids long) present at the N-terminus of newly synthesized proteins that directs their entry to the secretory pathway.
  • the signal peptide is covalently joined to the 5’ terminus (N- terminus) of the extracellular domain.
  • the signal peptide comprises a CD8 polypeptide, e.g., the CAR comprises a truncated CD8 signal peptide.
  • the signal peptide is generated from the antibody from it is derived.
  • the signal peptide is linked to the N-terminus of a V H disclosed herein.
  • the signal peptide is linked to the N-terminus of a VL disclosed herein.
  • the signal peptide comprises or consists of the amino acid sequence set forth in SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 282, SEQ ID NO: 283, SEQ ID NO: 284, SEQ ID NO: 285, SEQ ID NO: 286, SEQ ID NO: 287, SEQ ID NO: 288, or SEQ ID NO: 289.
  • MGWSCIIFFLVATATGVLS (SEQ ID NO: 274) MKCSWVIFFLMAVVTGVNS (SEQ ID NO: 275) MNFGLRLIFLVLVLKGVLC (SEQ ID NO: 276) MGWSCIILFLVATATGVHS (SEQ ID NO: 277) MGWSCIIFFLVATATGVHS (SEQ ID NO: 278) MGWSSIILFLVATASGVHS (SEQ ID NO: 279) MAVLGLLFCLVTFPSCVLS (SEQ ID NO: 280) MSPAQFLFLSVLWIRETNG (SEQ ID NO: 281) MSPAQFLFLLVLWIREIHG (SEQ ID NO: 282) NAI-1540294631v3 140 MDSQAQVLMLLLLWVSGTCG (SEQ ID NO: 283) MSPAQFLFLLVLWNRETNG (SEQ ID NO: 284) MESHTQAFVFAFLWLSGVDG (SEQ ID NO: 285) MSPAQFLFLLVLWIRETNG (SEQ ID NO: 286)
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 275 is set forth in SEQ ID NO: 291.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 276 is set forth in SEQ ID NO: 292.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 277 is set forth in SEQ ID NO: 293 or SEQ ID NO: 294.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 278 is set forth in SEQ ID NO: 295.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 279 is set forth in SEQ ID NO: 296.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 280 is set forth in SEQ ID NO: 297.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 281 is set forth in SEQ ID NO: 298.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 282 is set forth in SEQ ID NO: 299.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 283 is set forth in SEQ ID NO: 300.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 284 is set forth in SEQ ID NO: 301.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 285 is set forth in SEQ ID NO: 302.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 286 is set forth in SEQ ID NO: 303 or SEQ ID NO: 304.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 287 is set forth in SEQ ID NO: 305.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 288 is set forth in SEQ ID NO: 306.
  • an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 289 is set forth in SEQ ID NO: 307.
  • SEQ ID NOs: 290-307 are provided below.
  • the signal peptide is linked to the N-terminus of the VH.
  • the extracellular domain of the CAR comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 47 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278.
  • the signal peptide is linked to the N-terminus of the V H .
  • the extracellular domain of the CAR comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 62 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278.
  • the signal peptide is linked to the N-terminus of the V H .
  • the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75 and a signal peptide comprising the amino acid NAI-1540294631v3 142 sequence set forth in SEQ ID NO: 278.
  • the signal peptide is linked to the N-terminus of the VH.
  • the extracellular domain of the CAR comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 89 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 274.
  • the signal peptide is linked to the N-terminus of the V H .
  • the extracellular domain of the CAR comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 101 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the V H .
  • the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280.
  • the signal peptide is linked to the N-terminus of the VH.
  • the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280.
  • the signal peptide is linked to the N-terminus of the VH.
  • the extracellular domain of the CAR comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 155 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 275.
  • the signal peptide is linked to the N-terminus of the V H .
  • the extracellular domain of the CAR comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 170 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280.
  • the signal peptide is linked to the N-terminus of the V H .
  • the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278.
  • the signal peptide is linked to the N-terminus of the V H .
  • the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201 and a signal peptide comprising the amino NAI-1540294631v3 143 acid sequence set forth in SEQ ID NO: 276.
  • the signal peptide is linked to the N-terminus of the VH.
  • the extracellular domain of the CAR comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 215 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280.
  • the signal peptide is linked to the N-terminus of the V H .
  • the extracellular domain of the CAR comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 233 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 279.
  • the signal peptide is linked to the N-terminus of the V H .
  • the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280.
  • the signal peptide is linked to the N-terminus of the VH.
  • the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 262 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 279. In certain embodiments, the signal peptide is linked to the N-terminus of the VH.
  • the extracellular domain of the CAR comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 30, and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 281. In certain embodiments, the signal peptide is linked to the N-terminus of the V L .
  • the extracellular domain of the CAR comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 48 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 286. In certain embodiments, the signal peptide is linked to the N-terminus of the V L . [00482] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 63 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 286. In certain embodiments, the signal peptide is linked to the N-terminus of the V L .
  • the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 76 and a signal peptide comprising the amino acid NAI-1540294631v3 144 sequence set forth in SEQ ID NO: 286.
  • the signal peptide is linked to the N-terminus of the VL.
  • the extracellular domain of the CAR comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 90 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 286.
  • the signal peptide is linked to the N-terminus of the V L .
  • the extracellular domain of the CAR comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 102 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 282. In certain embodiments, the signal peptide is linked to the N-terminus of the V L . [00486] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 120 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 287. In certain embodiments, the signal peptide is linked to the N-terminus of the VL.
  • the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 138 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 288. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00488] In certain embodiments, the extracellular domain of the CAR comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 156 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 283. In certain embodiments, the signal peptide is linked to the N-terminus of the V L .
  • the extracellular domain of the CAR comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 171 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 288. In certain embodiments, the signal peptide is linked to the N-terminus of the V L . [00490] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 184 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 284. In certain embodiments, the signal peptide is linked to the N-terminus of the V L .
  • the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 202 and a signal peptide comprising the amino NAI-1540294631v3 145 acid sequence set forth in SEQ ID NO: 285.
  • the signal peptide is linked to the N-terminus of the VL.
  • the extracellular domain of the CAR comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 216 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 288.
  • the signal peptide is linked to the N-terminus of the V L .
  • the extracellular domain of the CAR comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 234 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 289. In certain embodiments, the signal peptide is linked to the N-terminus of the V L . [00494] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 247 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 287. In certain embodiments, the signal peptide is linked to the N-terminus of the VL.
  • the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 263 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 289. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00496] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 31, and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 49 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00498] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 77 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 139 and a signal peptide comprising NAI-1540294631v3 146 the amino acid sequence set forth in SEQ ID NO: 278.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 157 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 264 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 32, and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 50 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00504] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 78 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 140 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00506] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 158 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 265 and a signal peptide comprising NAI-1540294631v3 147 the amino acid sequence set forth in SEQ ID NO: 278.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 31, and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 49 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00510] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 64 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 77 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 91 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 103 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 121 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 139 and a signal peptide comprising NAI-1540294631v3 148 the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR f comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 157 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 172 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 185 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 203 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 217 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 235 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 248 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv.
  • the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 264 and a signal peptide comprising NAI-1540294631v3 149 the amino acid sequence set forth in SEQ ID NO: 277.
  • the signal peptide is linked to the N-terminus of the scFv. 5.3.2. Transmembrane Domain of a CAR
  • the transmembrane domain of the CAR comprises a hydrophobic alpha helix that spans at least a portion of the membrane.
  • the transmembrane domain may be derived either from a natural or from a synthetic source. Where the source is natural, the domain may be derived from any membrane-bound or transmembrane protein.
  • the transmembrane region may be derived from (e.g., comprise at least the transmembrane region(s) of) the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8 (e.g., CD8 alpha, CD8 beta), CD9, CD16, CD22, CD27, CD33, CD37, CD40, CD64, CD80, CD86, CD134, CD137, CD154, ICAM-1, KIRDS2, OX40, CD2, NKGD2, LFA-1 (CD11a, CD18) , ICOS (CD278), 4-1BB (CD137), GITR, CD40, BAFFR, HVEM (LIGHTR) , SLAMF7, NKp80 (KLRF1), CD160, CD166, CD19, IL2R beta, IL2R gamma, IL7R ⁇ , ITGA1, VLA1, CD49a, ITGA4,
  • the transmembrane domain of the CAR comprises a native or modified transmembrane domain of CD8, of CD28, of CD3 ⁇ , of CD4, of 4-1BB, of OX40, of ICOS, of CD84, of CD166, of CD8a, of CD8b, of ICAM-1, of CTLA-4, of CD27, of CD40, of NKGD2, or a combination thereof.
  • the transmembrane domain may be synthetic, in which case it comprises predominantly hydrophobic residues such as leucine and valine. In some cases, a triplet of phenylalanine, tryptophan and valine can be found at each end of a synthetic transmembrane domain.
  • a short oligo- or polypeptide linker such as between 2 and 10 amino acids in length, may form the linkage between the transmembrane domain and the intracellular domain of the CAR.
  • the CAR comprises more than one transmembrane domain, which can be a repeat of the same transmembrane domain, or can be different transmembrane domains.
  • the transmembrane domain of the CAR comprises a CD28 polypeptide (e.g., a transmembrane domain of CD28 or a portion thereof).
  • NAI-1540294631v3 150 the transmembrane domain of the CAR comprises a transmembrane domain of human CD28 or a portion thereof.
  • the CD28 polypeptide can comprise or consist of an amino acid sequence that is at least about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99% or 100% identical or homologous to the sequence having a NCBI Reference No: NP_006130 (SEQ ID NO: 308), or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions.
  • the CD28 polypeptide comprises or consists of an amino acid sequence that is a consecutive portion of SEQ ID NO: 308, which is at least about 20, or at least about 30, or at least about 40, or at least about 50, or at least about 60, and/or up to about 70, up to about 80, up to about 90, up to about 100, up to about 150, up to about 200, or up to about 220 amino acids in length.
  • the CD28 polypeptide comprises or consists of the amino acid sequence of amino acids 1 to 220, 1 to 50, 50 to 100, 100 to 150, 154 to 179, 150 to 200, 153 to 179, or 200 to 220 of SEQ ID NO: 308.
  • the transmembrane domain of the CAR comprises a CD28 polypeptide comprising or consisting of the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308.
  • SEQ ID NO: 308 is provided below. MLRLLLALNLFPSIQVTGNKILVKQSPMLVAYDNAVNLSCKYSYNLFSREFRASLHKGLDSAVEVCV VYGNYSQQLQVYSKTGFNCDGKLGNESVTFYLQNLYVNQTDIYFCKIEVMYPPPYLDNEKSNGTIIH VKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGP TRKHYQPYAPPRDFAAYRS [SEQ ID NO: 308] [00528] An exemplary nucleotide sequence encoding the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308 is set forth in SEQ ID NO: 309,
  • the transmembrane domain of the CAR comprises a CD8 polypeptide (e.g., a transmembrane domain of CD8 or a portion thereof). In certain embodiments, the transmembrane domain of the CAR comprises a transmembrane domain of human CD8 or a portion thereof.
  • the CD8 polypeptide comprises or has an amino acid sequence that is at least about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the sequence having a NCBI Reference No: NP_001139345.1 (SEQ ID NO: 310) or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions.
  • the CD8 polypeptide comprises or consists of an amino acid sequence that is a consecutive portion of SEQ ID NO: 310, which is at least about 20, or at least about 30, or at least about 40, or at least NAI-1540294631v3 151 about 50, and up to about 235 amino acids in length.
  • the CD8 polypeptide comprises or consists of an amino acid sequence of amino acids 1 to 235, 1 to 50, 50 to 100, 100 to 150, 150 to 200, 137 to 209 or 200 to 235 of SEQ ID NO: 310.
  • the transmembrane domain of the CAR comprises a CD8 polypeptide comprising or consisting of the amino acid sequence of amino acids 137 to 209 of SEQ ID NO: 310. SEQ ID NO: 310 is provided below.
  • the CAR further comprise a spacer/hinge domain that links the extracellular domain to the transmembrane domain.
  • a hinge domain is a short sequence of amino acids that facilitates antibody flexibility (see, e.g., Woof et al., Nat. Rev. Immunol.4(2): 89-99 (2004)).
  • the hinge domain can be any suitable sequence derived or obtained from any suitable molecule.
  • the hinge domain can be flexible enough to allow the extracellular domain to orient in different directions to facilitate antigen recognition while preserving the activating activity of the CAR.
  • the hinge domain sequence is derived from a CD8a molecule or a CD28 molecule.
  • the hinge/spacer domain of the CAR comprises a native or modified hinge region of CD8, of CD28, of CD3 ⁇ , of CD40, of 4-1BB, of OX40, of CD84, of CD166, of CD8a, of CD8b, of ICOS, of ICAM-1, of CTLA-4, of CD27, of CD40, of NKGD2, or a combination thereof.
  • the hinge/spacer region can be the hinge region from IgG1, or the CH2CH3 region of immunoglobulin and portions of CD3, a portion of a CD28 polypeptide (e.g., a portion of SEQ ID NO: 308), a portion of a CD8 polypeptide (e.g., a portion of SEQ ID NO: 310), a variant of any of the foregoing which is at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100% identical or homologous thereto, or a synthetic spacer sequence.
  • the hinge domain of the CAR comprises a native or modified hinge region of CD28.
  • the hinge domain of the CAR comprises a native hinge region of CD28. In certain embodiments, the hinge domain of the CAR comprises the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308.
  • An exemplary nucleotide sequence encoding the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308 is set forth in SEQ ID NO: 311, which is provided below.
  • NAI-1540294631v3 152 ATTGAAGTTATGTATCCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGA AAGGGAAACACCTTTGTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTT [SEQ ID NO: 311] 5.3.3.
  • the CAR comprises an intracellular domain.
  • the intracellular domain of the CAR comprises a CD3 ⁇ polypeptide.
  • CD3 ⁇ can activate or stimulate a cell (e.g., an immune effector cell, e.g., a cell of the lymphoid lineage, e.g., a T-cell) after antigen recognition.
  • the intracellular domain of the CD3 ⁇ -chain is the primary transmitter of signals from endogenous TCRs.
  • the intracellular domain of the CAR transmits a signal to the immune effector cell expressing the CAR after antigen recognition, activating at least one of the normal effector functions of the immune effector cell.
  • the effector function of a T cell is cytolytic activity or helper activity, including secretion of cytokines.
  • the intracellular domain can comprise the “intracellular signaling domain” of a T cell receptor (TCR) and optional co-receptors. While usually the entire intracellular signaling domain can be employed, in many cases it is not necessary to use the entire chain. To the extent that a truncated portion of the intracellular signaling domain is used, such truncated portion may be used in place of the intact chain as long as it transduces the effector function signal.
  • Cytoplasmic signaling sequences that regulate primary activation of the TCR complex that act in a stimulatory manner may contain signaling motifs which are known as immunoreceptor tyrosine-based activation motifs (ITAMs).
  • ITAMs immunoreceptor tyrosine-based activation motifs
  • Examples of ITAM containing cytoplasmic signaling sequences include those derived from CD8, CD3 ⁇ , CD3 ⁇ , CD3 ⁇ , CD3 ⁇ , CD32 (Fc gamma RIIa), DAP10, DAP12, CD79a, CD79b, Fc ⁇ RI ⁇ , Fc ⁇ RIII ⁇ , Fc ⁇ RI ⁇ (FCERIB), and Fc ⁇ RI ⁇ (FCERIG).
  • the intracellular signaling domain is derived from CD3 zeta (CD3 ⁇ ) (TCR zeta, GenBank acc. no. BAG36664.1).
  • CD3 ⁇ T-cell surface glycoprotein CD3 zeta (CD3 ⁇ ) chain, also known as T-cell receptor T3 zeta chain or CD247 (Cluster of Differentiation 247), is a protein that in humans is encoded by the CD247 gene.
  • Wild type (“native”) CD3 ⁇ comprises three functional immunoreceptor tyrosine-based activation motifs (ITAMs), three functional basic-rich stretch (BRS) regions (BRS1, BRS2 and BRS3).
  • ITAMs immunoreceptor tyrosine-based activation motifs
  • BRS basic-rich stretch
  • the intracellular tails of the CD3 molecules comprise a single ITAM, which is responsible for the signaling capacity of the TCR.
  • the intracellular tail of the ⁇ chain (CD3 ⁇ ) comprises three (3) ITAMs.
  • the intracellular domain of the CAR comprises a native CD3 ⁇ .
  • the native CD3 ⁇ comprises or consists of an amino acid sequence that is at NAI-1540294631v3 153 least about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence having a NCBI Reference No: NP_932170 (SEQ ID NO: 312), or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions.
  • the CD3 ⁇ polypeptide comprises or consists of an amino acid sequence that is a consecutive portion of SEQ ID NO: 312, which is at least about 20, or at least about 30, or at least about 40, or at least about 50, and up to about 164 amino acids in length.
  • the native CD3 ⁇ comprises or consists of the amino acid sequence of amino acids 1 to 164, 1 to 50, 50 to 100, 52 to 164, 100 to 150, or 150 to 164 of SEQ ID NO: 312.
  • the intracellular signaling domain of the CAR comprises a native CD3 ⁇ comprising or consisting of the amino acid sequence of amino acids 52 to 164 of SEQ ID NO: 312. SEQ ID NO: 312 is provided below.
  • the intracellular domain of the CAR comprises a modified CD3 ⁇ polypeptide.
  • the modified CD3 ⁇ polypeptide comprises a mutation, such as a point mutation, in at least one ITAM so as to render the ITAM non-functional.
  • the modified CD3 ⁇ polypeptide is missing either two membrane-proximal ITAMs (ITAM1 and ITAM2) or two membrane-distal ITAMs (ITAM2 and ITAM3).
  • the modified CD3 ⁇ polypeptide is missing ITAM2. Removing at least one ITAM from the introduced CAR may reduce CD3 ⁇ -mediated apoptosis. Alternatively, removing at least one ITAM from the introduced CAR can reduce its size without loss of function.
  • the modified CD3 ⁇ polypeptide comprises one, two or three ITAMs. In certain embodiments, the modified CD3 ⁇ polypeptide comprises a native ITAM1.
  • the native ITAM1 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 313.
  • NAI-1540294631v3 154 QNQLYNELNLGRREEYDVLDKR [SEQ ID NO: 313]
  • An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 313 is set forth in SEQ ID NO: 314, which is provided below.
  • the modified CD3 ⁇ polypeptide comprises an ITAM1 variant comprising one or more loss-of-function mutations.
  • the modified CD3 ⁇ polypeptide comprises a native ITAM2.
  • the native ITAM2 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 317, which is provided below.
  • QEGLYNELQKDKMAEAYSEIGMK [SEQ ID NO: 317]
  • An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 317 is set forth in SEQ ID NO: 318, which is provided below.
  • the modified CD3 ⁇ polypeptide comprises an ITAM2 variant.
  • the ITAM2 variant comprises or consists of one or more loss-of-function mutations.
  • the ITAM2 variant comprises or consists of two loss-of-function mutations.
  • each of the one or more (e.g., two) the loss of function mutations comprises a mutation of a tyrosine residue in ITAM2.
  • the ITAM1 variant consists of two loss-of-function mutations.
  • the ITAM2 variant comprises or consists of the amino acid sequence set forth in SEQ ID NO: 319, which is provided below.
  • QEGLFNELQKDKMAEAFSEIGMK [SEQ ID NO: 319] NAI-1540294631v3 155 [00547]
  • An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 319 is set forth in SEQ ID NO: 320, which is provided below.
  • the modified CD3 ⁇ polypeptide comprises a native ITAM3.
  • the native ITAM3 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 321, which is provided below.
  • HDGLYQGLSTATKDTYDALHMQ [SEQ ID NO: 321]
  • An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 321 is set forth in SEQ ID NO: 322, which is provided below.
  • CACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAG [SEQ ID NO: 322]
  • the modified CD3 ⁇ polypeptide comprises an ITAM3 variant.
  • the ITAM3 variant comprises or consists of two loss-of-function mutations.
  • each of the one or more (e.g., two) the loss of function mutations comprises a mutation of a tyrosine residue in ITAM3.
  • the ITAM3 variant comprises or consists of two loss-of-function mutations.
  • the ITAM3 variant comprises or consists of the amino acid sequence set forth in SEQ ID NO: 323, which is provided below.
  • the intracellular domain of the CAR comprises a modified CD3 ⁇ polypeptide comprising a native ITAM1, an ITAM2 variant comprising or consisting of one or more (e.g., two) loss-of-function mutations, and an ITAM3 variant comprising or consisting of one or more (e.g., two) loss-of-function mutations.
  • the intracellular domain of the CAR comprises a modified CD3 ⁇ polypeptide comprising a native ITAM1, an ITAM2 variant consisting of two loss-of-function mutations, and an ITAM3 variant consisting of two loss-of- function mutations.
  • the intracellular domain of the CAR comprises a NAI-1540294631v3 156 modified CD3 ⁇ polypeptide comprising a native ITAM1 consisting of the amino acid sequence set forth in SEQ ID NO: 313, an ITAM2 variant consisting of the amino acid sequence set forth in SEQ ID NO: 319, and an ITAM3 variant consisting of the amino acid sequence set forth in SEQ ID NO: 323.
  • the modified CD3 ⁇ polypeptide comprises or consists of the amino acid sequence set forth in SEQ ID NO: 325. SEQ ID NO: 325 is provided below.
  • a CAR comprising an intracellular domain comprising a modified CD3 ⁇ polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 325 is designated as an “1XX” CAR.
  • the intracellular domain of the CAR comprises a modified CD3 ⁇ polypeptide comprising or consisting of an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, at least about 100% identical to SEQ ID NO: 43 or a fragment thereof, and/or may optionally comprise up to one or up to two or up
  • SEQ ID NO: 326 An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 325 is set forth in SEQ ID NO: 326, which is provided below.
  • the intracellular domain of the CAR further comprises at least one co-stimulatory signaling
  • the at least one co-stimulatory region comprises a co-stimulatory molecule or a portion thereof. In certain embodiments, the at least one co-stimulatory region comprises an intracellular domain of at least one co-stimulatory molecule or a portion thereof.
  • a “co-stimulatory molecule” refers to a cell surface molecule other than antigen receptor or its ligand that can provide an efficient response of lymphocytes to an antigen. A co-stimulatory molecule can provide optimal lymphocyte activation.
  • the co-stimulatory molecule can bind to a co-stimulatory ligand, which is a protein expressed on cell surface that upon binding to its receptor produces a co-stimulatory response, i.e., an intracellular response that effects the NAI-1540294631v3 157 stimulation provided when an antigen-recognizing receptor (e.g., a chimeric antigen receptor (CAR)) binds to its target antigen.
  • a 4-1BB ligand i.e., 4-1BBL
  • 4-1BB may bind to 4-1BB for providing an intracellular signal that in combination with a CAR signal induces an effector cell function of the CAR + T-cell.
  • Non-limiting examples of co-stimulatory molecules include CD28, 4-1BB (CD137), OX40, ICOS, DAP-10, CD27, CD28, CD30, CD40, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, CD8, CD4, B2C, CD80, CD86, DAP10, DAP12, MyD88, BTNL3, and NKG2D. and combinations thereof.
  • LFA-1 lymphocyte function-associated antigen-1
  • the intracellular domain of the CAR comprises a co-stimulatory signaling region that comprises a CD28 polypeptide, e.g., an intracellular domain of CD28 or a portion thereof. In certain embodiments, the intracellular domain of the CAR comprises a co- stimulatory signaling region that comprises an intracellular domain of human CD28 or a portion thereof.
  • the CD28 polypeptide comprised in the co-stimulatory signaling region of the CAR comprise or consists of an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, at least about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 308, or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions.
  • the CD28 polypeptide comprised in the co-stimulatory signaling region comprises or consist of an amino acid sequence that is a consecutive portion of SEQ ID NO: 308, which is at least about 20, or at least about 30, or at least about 40, or at least about 50, and up to about 220 amino acids in length.
  • the CD28 polypeptide comprised in the co- stimulatory signaling region comprises or consists of amino acids 1 to 220, 1 to 50, 50 to 100, 100 to 150, 114 to 220, 150 to 200, 180 to 220, or 200 to 220 of SEQ ID NO: 308.
  • the intracellular signaling domain of the CAR comprises a co-stimulatory signaling region that comprises a CD28 polypeptide comprising or consisting of the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308.
  • a co-stimulatory signaling region that comprises a CD28 polypeptide comprising or consisting of the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308.
  • An exemplary nucleic acid sequence encoding the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308 is set forth in SEQ ID NO: 327, which is provided below.
  • the intracellular domain of the CAR comprises a modified CD28 domain that imparts unique functional properties to the CAR.
  • a native CD28 domain comprises three intracellular subdomains consisting of the amino acid sequences YMNM, PRRP, and PYAP that regulate signaling pathways post stimulation.
  • the intracellular domain of the CAR comprises a modified CD28 domain wherein one or more of the YMNM, PRRP, and PYAP subdomains are mutated or deleted, so as to amplify, attenuate, or inactivate said subdomain(s), thereby modulating CAR-T function.
  • the intracellular domain of the CAR comprises a modified CD28 domain disclosed in International Patent Application Publication Nos.: WO 2019/010383 and WO 2021158850, which are incorporated by reference in their entireties.
  • the intracellular domain of the CAR comprises a co-stimulatory signaling region that comprises a 4-1BB polypeptide, e.g., an intracellular domain of 4-1BB or a portion thereof.
  • the co-stimulatory signaling region comprises an intracellular domain of human 4-1BB or a portion thereof.
  • the 4-1BB comprised in the co-stimulatory signaling region comprises or consists of an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, at least about 100% identical or homologous to the sequence having a NCBI Ref No.: NP_001552 (SEQ ID NO: 328) or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions.
  • the 4-1BB comprised in the co-stimulatory signaling region comprises or consists of an amino acid sequence that is a consecutive portion of SEQ ID NO: 328, which is at least about 20, or at least about 30, or at least about 40, or at least about 50, and/or up to about 50, up to about 60, up to about 70, up to about 80, up to about 90, up to about 100, up to about 200, or up to about 255 amino acids in length.
  • the 4-1BB polypeptide comprised in the co-stimulatory signaling region comprises or consists of the amino acid sequence of amino acids 1 to 255, 1 to 50, 50 to 100, 100 to 150, 150 to 200, or 200 to 255 of SEQ ID NO: 328.
  • the co-stimulatory signaling region comprises a 4-1BB polypeptide comprising or consisting of the amino acid sequence of amino acids 214 to 255 of SEQ ID NO: 328.
  • SEQ ID NO: 328 is provided below.
  • NAI-1540294631v3 159 MGNSCYNIVATLLLVLNFERTRSLQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQ CKGVFRTRKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQELTKKGCKDCCFGTFNDQKRGICR PWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSVTPPAPAREPGHSPQIISFFLALTSTALLF LLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL [SEQ ID NO: 328] [00564]
  • the intracellular domain of the CAR comprises a co-stimulatory signaling region
  • the intracellular domain of the CAR further comprises a peptide.
  • the self-cleaving P2A peptide comprises or consists of the amino acid sequence of SEQ ID NO: 329.
  • SEQ ID NO: 329 is provided below.
  • GSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 329]
  • An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 329 is set forth in SEQ ID NO: 330, which is provided below.
  • the CAR is expressed with a C-terminal LNGFR tag.
  • the LNGFR tag comprises or consists of the amino acid sequence of SEQ ID NO: 331. SEQ ID NO: 331 is provided below.
  • SEQ ID NO: 331 An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 331 is set forth in SEQ ID NO: 332, which is provided below.
  • the CAR comprises (a) an extracellular antigen-binding domain comprising (i) a V H that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 25, and (ii) a VL that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 28, (b) a hinge domain comprising a CD28 polypeptide (e.g., a hinge domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 30
  • the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329.
  • the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278).
  • the V H and VL are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00570]
  • the VH and VL are positioned from the N- to the C-terminus: VH- V L .
  • the CAR is designed as “1_28z1XX (vH-vK)” or “ATA017 VH-VK CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 333, which is provided below.
  • NAI-1540294631v3 161 MGWSCIIFFLVATATGVHSQVQLQQSGPEVVRPGVSVKISCKGSGYKFTDYAMHWVKQSHAKSLEWI GVISTYNGNTNYNRKFKDKATMTVDKSSSTAYMELARLTSEDSAIYYCLKNDYWGQGTSVTVSSGGG GSGGGGSGGGGSDVVMTQTPLTLSVTIGQPASFSCKSSQSLLDSDGKTYLNWVLQRPGQSPNRLIYL VSKLDSRVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTQFPHTFGGGTKLEIKRAAAIEVMYP PPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRL LHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEY
  • the CAR is designed as “1_28z1XX (vK-vH)” or “ATA017 VK-VH CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 335, which is provided below.
  • the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329.
  • the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278).
  • the VH and VL are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00575]
  • the V H and V L are positioned from the N- to the C-terminus: V H - V L .
  • the CAR is designed as “2_28z1XX (vH-vK)” or “ATA018 VH-VK CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 337, which is provided below.
  • the CAR is designed as “2_28z1XX (vK-vH)” or “ATA018 VK-VH CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 339, which is provided below.
  • the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329.
  • the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278).
  • the V H and V L are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00580]
  • the VH and VL are positioned from the N- to the C-terminus: VH- V L .
  • the CAR is designed as “4_28z1XX (vH-vK)” or “ATA019 VH-VK CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 341, which is provided below.
  • the CAR is designed as “4_28z1XX (vK-vH)” or “ATA019 VK-VH CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 343, which is provided below.
  • the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329.
  • the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278).
  • the V H and V L are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00585]
  • the VH and VL are positioned from the N- to the C-terminus: VH- V L .
  • the CAR is designed as “8_28z1XX (vH-vK)” or “ATA020 VH-VK CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 345, which is provided below.
  • the CAR is designed as “8_28z1XX (vK-vH)” or “ATA020 VK-VH CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 347, which is provided below.
  • the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329.
  • the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278).
  • the VH and VL are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00590]
  • the V H and V L are positioned from the N- to the C-terminus: V H - VL.
  • the CAR is designed as “9_28z1XX (vH-vK)” or “ATA021 VH-VK CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 349, which is provided below.
  • the CAR is designed as “9_28z1XX (vK-vH)” or “ATA021 VK-VH CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 351, which is provided below.
  • the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329.
  • the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278).
  • the V H and V L are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00595]
  • the VH and VL are positioned from the N- to the C-terminus: VH- V L .
  • the CAR is designed as “16_28z1XX (vH-vK)” or “ATA022 VH-VK CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 353, which is provided below.
  • the CAR is designed as “16_28z1XX (vK-vH)” or “ATA022 VK-VH CAR”.
  • the CAR comprises the amino acid sequence set forth in SEQ ID NO: 355, which is provided below.
  • the presently disclosed subject matter provides cells comprising a presently disclosed GPC3-targeted CAR (e.g., one disclosed in Section 5.3).
  • the cell is selected from the group consisting of cells of lymphoid lineage and cells of myeloid lineage.
  • the cell is an immunoresponsive cell.
  • the immunoresponsive cell is a cell of lymphoid lineage.
  • the cell is a cell of the lymphoid lineage. Cells of the lymphoid lineage can provide production of antibodies, regulation of cellular immune system, detection of foreign agents in the blood, detection of cells foreign to the host, and the like.
  • Non-limiting examples of cells of the lymphoid lineage include T cells, Natural Killer (NK) cells, B cells, NAI-1540294631v3 178 dendritic cells, and stem cells from which lymphoid cells may be differentiated.
  • the stem cell is a pluripotent stem cell (e.g., embryonic stem cell or an induced pluripotent stem cell).
  • the cell is an immune effector cell. Immune effector cells can be obtained from a number of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors.
  • Immune effector cells can be obtained from blood collected from a subject using any number of techniques known to the skilled artisan, such as FicollTM separation.
  • cells from the circulating blood of an individual may be obtained by apheresis.
  • immune effector cells are isolated from peripheral blood lymphocytes by lysing the red blood cells and depleting the monocytes, for example, by centrifugation through a PERCOLLTM gradient or by counterflow centrifugal elutriation.
  • a specific subpopulation of immune effector cells can be further isolated by positive or negative selection techniques.
  • immune effector cells can be isolated using a combination of antibodies directed to surface markers unique to the positively selected cells, e.g., by incubation with antibody-conjugated beads for a time period sufficient for positive selection of the desired immune effector cells.
  • enrichment of immune effector cells population can be accomplished by negative selection using a combination of antibodies directed to surface markers unique to the negatively selected cells.
  • the immune effector cells comprise any leukocyte involved in defending the body against infectious disease and foreign materials.
  • the immune effector cells can comprise lymphocytes, monocytes, macrophages, dendritic cells, mast cells, neutrophils, basophils, eosinophils, or any combinations thereof.
  • the immune effector cells can comprise T lymphocytes, preferably cytotoxic T lymphocytes (CTLs).
  • TTLs cytotoxic T lymphocytes
  • the cell is a T cell.
  • T cells can be lymphocytes that mature in the thymus and are chiefly responsible for cell-mediated immunity. T cells are involved in the adaptive immune system. T cells or T lymphocytes can be distinguished from other lymphocytes, such as B cells and natural killer cells (NK cells), by the presence of a T-cell receptor (TCR) on the cell surface. They are called T cells because they mature in the thymus (although some also mature in the tonsils). There are several subsets of T cells, each with a distinct function.
  • the T cells of the presently disclosed subject matter can be any type of T cells, including, but not limited to, helper T cells, cytotoxic T cells, memory T cells (including central memory T cells, stem-cell-like memory T cells (or stem-like memory T cells), and two types of effector memory T cells: e.g., T EM cells and T EMRA cells), Regulatory T cells (also known as suppressor T NAI-1540294631v3 179 cells or Treg), tumor-infiltrating lymphocyte (TIL), Natural killer T cells (NK-T cells), mucosal associated invariant T cells, ⁇ T cells, and ⁇ T cells.
  • helper T cells include cytotoxic T cells, memory T cells (including central memory T cells, stem-cell-like memory T cells (or stem-like memory T cells), and two types of effector memory T cells: e.g., T EM cells and T EMRA cells), Regulatory T cells (also known as suppressor T NAI-1540294631v3 179 cells or Treg
  • the cell comprising the presently disclosed GPC3-targeted CAR is a helper T cell or CD4 + T cell.
  • Helper T cells or T helper cells, or TH cells
  • helper T cells assist other white blood cells in immunologic processes, including maturation of B cells into plasma cells and memory B cells, and activation of cytotoxic T cells and macrophages. These cells are also known as CD4 + T cells because they express the CD4 glycoprotein on their surface.
  • Helper T cells become activated when they are presented with peptide antigens by MHC class II molecules, which are expressed on the surface of antigen-presenting cells (APCs). Once activated, they divide rapidly and secrete small proteins called cytokines that regulate or assist in the active immune response.
  • APCs antigen-presenting cells
  • the cell comprising the presently disclosed GPC3-targeted CAR is a cytotoxic T cell or CD8 + T cell.
  • Cytotoxic T cells TC cells, or CTLs
  • TC cells destroy virally infected cells and tumor cells and are also implicated in transplant rejection.
  • CD8 + T cells since they express the CD8 glycoprotein at their surface. These cells recognize their targets by binding to antigen associated with MHC class I molecules, which are present on the surface of all nucleated cells.
  • the cell comprising the presently disclosed GPC3-targeted CAR is a memory T cell.
  • Memory T cells are a subset of antigen-specific T cells that persist long-term after an infection has resolved. They quickly expand to large numbers of effector T cells upon re- exposure to their cognate antigen, thus providing the immune system with “memory” against past infections. Memory cells may be either CD4 + or CD8 + . Memory T cells typically express the cell surface protein CD45RO.
  • the cell comprising the presently disclosed GPC3-targeted CAR is a regulatory T cell.
  • Regulatory T cells formerly known as suppressor T cells, are crucial for the maintenance of immunological tolerance. Their major role is to shut down T cell- mediated immunity toward the end of an immune reaction and to suppress auto-reactive T cells that escaped the process of negative selection in the thymus.
  • Two major classes of CD4 + T reg cells have been described — naturally occurring Treg cells and adaptive Treg cells.
  • the cell comprising the presently disclosed GPC3-targeted CAR is a Natural killer T cell.
  • Natural killer T (NKT) cells (not to be confused with natural killer (NK) NAI-1540294631v3 180 cells) bridge the adaptive immune system with the innate immune system.
  • NKT cells recognize glycolipid antigen presented by a molecule called CD1d.
  • the cell comprising the presently disclosed GPC3-targeted CAR is a ⁇ T cell.
  • ⁇ T cells possess a distinct T-cell receptor (TCR) having one ⁇ chain and one ⁇ chain instead of ⁇ and ⁇ chains.
  • the cell comprising the presently disclosed GPC3-targeted CAR is an innate lymphoid cell (ILC), a cytokine induced killer (CIK) cell, or a lymphokine activated killer (LAK) cell.
  • the cell is a T cell, and the presently disclosed GPC3-targeted CAR is integrated at a locus within the genome of the T cell.
  • the loci include a TRAC locus, a TRBC locus, a TRDC locus, and a TRGC locus.
  • the locus is a TRAC locus or a TRBC locus.
  • the T cells comprise a mixture of CD4 + T cells and CD8 + T cells.
  • the T cells are enriched for one or more subsets based on cell surface marker expression profile.
  • the cell is a Natural killer cell. Natural killer (NK) cells can be lymphocytes that are part of cell-mediated immunity and act during the innate immune response.
  • NK cells do not require prior activation in order to perform their cytotoxic effect on target cells. Unlike cytotoxic CD8 + T cells, NK cells launch cytotoxicity against tumor cells without the requirement for prior sensitization and can also eradicate MHC-I-negative cells.
  • the cell is a genetically modified NK cell. In certain embodiments, the cell is an edited NK cell. In certain embodiments, the cell is a NK cell derived from a stem cell. In certain embodiments, the cell is a NK cell derived from a pluripotent stem cell. In certain embodiments, the cell is an induced pluripotent stem cell (iPSC)-derived NK cell.
  • iPSC induced pluripotent stem cell
  • the presently disclosed cells can be autologous, non- autologous (e.g., allogeneic), or derived in vitro from engineered progenitor or stem cells.
  • the cells comprising a presently disclosed GPC3-targeted CAR are obtained from the subject to be treated (i.e. are autologous).
  • the cells comprising a presently disclosed GPC3-targeted CAR are allogeneic to the subject to be treated.
  • NAI-1540294631v3 181 [00616]
  • the cells of the presently disclosed subject matter can be cells of the myeloid lineage.
  • Epstein-Barr virus (EBV)-induced lymphoproliferative diseases (EBV-LPDs) and other EBV-associated cancers are a significant cause of morbidity and mortality for recipients of allogeneic hematopoietic cell transplantation (HCT) or solid organ transplants (SOT), particularly in those who have received certain T-cell reactive Abs to prevent or treat GVHD.
  • HCT allogeneic hematopoietic cell transplantation
  • SOT solid organ transplants
  • Prophylaxis and treatment by the adoptive transfer of autologous or allogeneic EBV-specific cytotoxic T cells and the subsequent long-term restoration of immunity against EBV-associated lymphoproliferation have provided positive outcomes in the management of these uniformly fatal complications of allogeneic tissue transfer.
  • the disclosed immune effector cells that comprise one or more of the CAR polypeptides of the present invention are allogeneic or autologous EBV- specific cytotoxic T lymphocytes (CTLs).
  • CTLs EBV-specific cytotoxic T lymphocytes
  • generation of EBV-specific cytotoxic T cells may involve isolating PBMCs from of an EBV-seropositive autologous or allogenic donor and enriching them for T cells by depletion of monocytes and NK cells.
  • EBV-specific cytotoxic T cells may also be produced by contacting donor PBMCs or purified donor T cells with a “stimulator” cell that expresses one or more EBV antigen(s) and presents the EBV antigen(s) to unstimulated T cells, thereby causing stimulation and expansion of EBV-specific CTLs.
  • EBV antigens include, for example, latent membrane protein (LMP) and EBV nuclear antigen (EBNA) proteins, such as LMP- 1, LMP-2A, and LMP-2B and EBNA-1, EBNA-2, EBNA-3A, EBNA-3B, EBNA-3C and EBNA- LP.
  • LMP latent membrane protein
  • EBNA EBV nuclear antigen
  • Cytotoxic T cells that comprise T cell receptor(s) which recognize one or more EBV-specific antigens are deemed to have been “sensitized” to those EBV antigen(s) and are therefore termed “EBV-sensitized cytotoxic T cells” herein.
  • the T cells used for generating the CAR-T cells of the invention are polyfunctional T- cells, i.e., those T cells that are capable of inducing multiple immune effector functions, that provide a more effective immune response to a pathogen than do cells that produce, for example, only a single immune effector (e.g. a single biomarker such as a cytokine or CD107a). Less-polyfunctional, monofunctional, or even “exhausted” T cells may dominate immune responses during chronic infections, thus negatively impacting protection against virus-associated complications.
  • the presently disclosed CAR-T cells are polyfunctional. In certain embodiments, at least 50% of the T cells used for generating the presently disclosed CAR-T cells are CD4 + T cells.
  • the T cells are less than 50% CD4 + T cells. In certain embodiments, the T cells are predominantly CD4 + T cells. In certain embodiments, at least 50% of the T cells used for generating the presently disclosed CAR-T cells are CD8 + T cells. In certain embodiments, the T cells are less than 50% CD8 + T cells. In certain embodiments, the T cells are predominantly CD8 + T cells. In some embodiments, the T cells (e.g., the sensitized T cells and/or CAR-T cells described herein) are stored in a cell library or bank before they are administered to the subject.
  • the cells can be transduced with the presently disclosed GPC3- targeted CAR such that the cells express the GPC3-targeted CAR.
  • Anti-GPC3 Antibodies [00619] The presently disclosed subject matter provides anti-GPC3 antibodies and antigen- binding fragments thereof.
  • the antibodies can be polyclonal antibodies, monoclonal antibodies, humanized antibodies, human antibodies, multispecific antibodies, bispecific antibodies, or hetero- conjugate antibodies, as well as variants thereof having increased or decreased affinity or other properties.
  • the presently disclosed anti-GPC3 antibody and antigen-binding fragment thereof binds to human GPC3. 5.5.1.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (V H ) and a light chain variable region (V L ).
  • V H comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a V H comprising the amino acid sequence of SEQ ID NO: 29; and/or ii) the V L comprises NAI-1540294631v3 183 a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 30.
  • the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 47; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 48.
  • SEQ ID NO: 47 and SEQ ID NO: 48 are disclosed in Table 2.
  • the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 62; and/or ii) the VL comprises a V L CDR1, a V L CDR2, and a V L CDR3 as set forth in a V L comprising the amino acid sequence of SEQ ID NO: 63.
  • SEQ ID NO: 62 and SEQ ID NO: 63 are disclosed in Table 3.
  • the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 75; and/or ii) the VL comprises a V L CDR1, a V L CDR2, and a V L CDR3 as set forth in a V L comprising the amino acid sequence of SEQ ID NO: 76.
  • SEQ ID NO: 75 and SEQ ID NO: 76 are disclosed in Table 4.
  • the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a V H comprising the amino acid sequence of SEQ ID NO: 89; and/or ii) the V L comprises a V L CDR1, a V L CDR2, and a V L CDR3 as set forth in a V L comprising the amino acid sequence of SEQ ID NO: 90.
  • SEQ ID NO: 89 and SEQ ID NO: 90 are disclosed in Table 5.
  • the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 as set forth in a V H comprising the amino acid sequence of SEQ ID NO: 101; and/or ii) the V L comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 102.
  • SEQ ID NO: 101 and SEQ ID NO: 102 are disclosed in Table 6.
  • the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 119; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 120.
  • SEQ ID NO: 119 and SEQ ID NO: 120 are disclosed in Table 7.
  • the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 137; and/or ii) the VL comprises a V L CDR1, a V L CDR2, and a V L CDR3 as set forth in a V L comprising the amino acid sequence of SEQ ID NO: 138.
  • SEQ ID NO: 137 and SEQ ID NO: 138 are disclosed in Table 8.
  • the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a V H comprising the amino acid sequence of SEQ ID NO: 155; and/or ii) the V L NAI-1540294631v3 184 comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 156.
  • SEQ ID NO: 155 and SEQ ID NO: 156 are disclosed in Table 9.
  • the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 170; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 171.
  • SEQ ID NO: 170 and SEQ ID NO: 171 are disclosed in Table 10.
  • the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 183; and/or ii) the VL comprises a V L CDR1, a V L CDR2, and a V L CDR3 as set forth in a V L comprising the amino acid sequence of SEQ ID NO: 184.
  • SEQ ID NO: 183 and SEQ ID NO: 184 are disclosed in Table 11.
  • the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 201; and/or ii) the VL comprises a V L CDR1, a V L CDR2, and a V L CDR3 as set forth in a V L comprising the amino acid sequence of SEQ ID NO: 202.
  • SEQ ID NO: 201 and SEQ ID NO: 202 are disclosed in Table 12.
  • the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a V H comprising the amino acid sequence of SEQ ID NO: 215; and/or ii) the V L comprises a V L CDR1, a V L CDR2, and a V L CDR3 as set forth in a V L comprising the amino acid sequence of SEQ ID NO: 216.
  • SEQ ID NO: 215 and SEQ ID NO: 216 are disclosed in Table 13.
  • the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 as set forth in a V H comprising the amino acid sequence of SEQ ID NO: 233; and/or ii) the V L comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 234.
  • SEQ ID NO: 233 and SEQ ID NO: 234 are disclosed in Table 14.
  • the V H comprises a V H CDR1, a V H CDR2, and a V H CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 246; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 247.
  • SEQ ID NO: 246 and SEQ ID NO: 247 are disclosed in Table 15.
  • the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 262; and/or ii) the VL comprises a V L CDR1, a V L CDR2, and a V L CDR3 as set forth in a V L comprising the amino acid sequence of SEQ ID NO: 263.
  • SEQ ID NO: 262 and SEQ ID NO: 263 are disclosed in Table 16.
  • the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 can be identified according to any known CDR numbering systems, which include but are not limited to, Kabat numbering system, Chothia numbering system, AbM numbering system. Contact numbering NAI-1540294631v3 185 system, and IMGT ® Information System.
  • the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28.
  • the CDRs are identified according to the Kabat numbering system.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification NAI-1540294631v3 186 thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the CDRs are identified according to the Kabat numbering system.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and a V L comprising a NAI-1540294631v3 187 CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61 SEQ ID NO: 26, SEQ ID NO: 27, and SEQ ID NO: 46 are disclosed in Table 3.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the CDRs are identified according to the Kabat numbering system.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set NAI-1540294631v3 188 forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a V L comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the CDRs are identified according to the Kabat numbering system.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43.
  • the V L comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, and SEQ ID NO: 46 are disclosed in Table 6.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, and SEQ ID NO: 118 are disclosed in Table 7.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, and SEQ ID NO: 136 are disclosed in Table 8.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: NAI-1540294631v3 191 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154.
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, and SEQ ID NO: 154 are disclosed in Table 9.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence NAI-1540294631v3 192 set forth in SEQ ID NO: 169.
  • the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 131, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 134, SEQ ID NO: 135, and SEQ ID NO: 169 are disclosed in Table 10. [00659]
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46.
  • the CDRs are identified according to the Kabat numbering system.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification thereof, and a CDR3 NAI-1540294631v3 193 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200.
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, and SEQ ID NO: 200 are disclosed in Table 12.
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and NAI-1540294631v3 194 a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169.
  • the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 213, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 214, SEQ ID NO: 135, and SEQ ID NO: 169 are disclosed in Table 13. [00665]
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232.
  • the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 227, SEQ ID NO: 228, SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, and SEQ ID NO: 232 are disclosed in Table 14. [00667]
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification NAI-1540294631v3 195 thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118.
  • the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 113, SEQ ID NO: 245, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, and SEQ ID NO: 118 are disclosed in Table 15. [00669]
  • the V H comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof.
  • the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260.
  • the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
  • the CDRs are identified according to the Kabat numbering system.
  • SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 230, SEQ ID NO: 231, and SEQ ID NO: 261 are disclosed in Table 16.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 29; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 30.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 29. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 30. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 29 and a V L comprising the amino acid sequence set forth in SEQ ID NO: 30.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 47; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 48.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 47. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a V L comprising the amino acid sequence set forth in SEQ ID NO: 48. In certain embodiments, the anti-GPC3 antibody or antigen- NAI-1540294631v3 197 binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 47 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 48.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 62; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 63.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 62. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 63. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 62 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 63.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 75; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 76.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 75. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 76. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 76.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 89; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at NAI-1540294631v3 198 least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 90.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 89. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 90. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 89 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 90.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 101; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 102.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 101. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 102. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 101 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 102.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 119; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 120.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 119. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 120. In certain embodiments, the anti-GPC3 antibody or antigen- NAI-1540294631v3 199 binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 120.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 137; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 138.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 137. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 138. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 138.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 155; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 156.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 155. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 156. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 156.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 170; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at NAI-1540294631v3 200 least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 171.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 170. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 171. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 170 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 171.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 183; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 184.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 183. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 184. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 183 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 184.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 201; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 202.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 201. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 202. In certain embodiments, the anti-GPC3 antibody or antigen- NAI-1540294631v3 201 binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 202.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 215; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 216.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 215. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 216. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 215 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 216.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 233; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 234.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 233. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 234. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 233 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 234.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 246; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at NAI-1540294631v3 202 least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 247.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 246. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 247. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 246 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 247.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 262; and/or a V L comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 263.
  • the anti-GPC3 antibody or antigen-binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 262. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 263. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 262 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 263.
  • the antibody designated as “ATA001” comprises the V H CDR1, V H CDR2, and V H CDR3 sequences disclosed in Table 1; and the V L CDR1, V L CDR2, and V L CDR3 sequences disclosed in Table 1.
  • the ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 29 (disclosed in Table 1) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 30 (disclosed in Table 1).
  • the antibody designated as “ATA002” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 2; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 2.
  • the ATA001 antibody comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 47 (disclosed in Table 2) and a V L comprising the amino acid sequence set forth in SEQ ID NO: 48 (disclosed in Table 2).
  • the antibody designated as “ATA003” comprises the V H CDR1, V H CDR2, and V H CDR3 sequences disclosed in Table 3; and the V L CDR1, V L CDR2, and V L CDR3 sequences NAI-1540294631v3 203 disclosed in Table 3.
  • the ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 62 (disclosed in Table 3) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 63 (disclosed in Table 3).
  • the antibody designated as “ATA004” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 4; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 4.
  • the ATA001 antibody comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 75 (disclosed in Table 4) and a V L comprising the amino acid sequence set forth in SEQ ID NO: 76 (disclosed in Table 4).
  • the ATA001 antibody comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 101 (disclosed in Table 6) and a V L comprising the amino acid sequence set forth in SEQ ID NO: 102 (disclosed in Table 6).
  • the antibody designated as “ATA007” comprises the V H CDR1, V H CDR2, and V H CDR3 sequences disclosed in Table 7; and the V L CDR1, V L CDR2, and V L CDR3 sequences disclosed in Table 7.
  • the ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119 (disclosed in Table 7) and a V L comprising the amino acid sequence set forth in SEQ ID NO: 120 (disclosed in Table 7).
  • the antibody designated as “ATA008” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 8; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 8.
  • the ATA001 antibody comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 137 (disclosed in Table 8) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 138 (disclosed in Table 8).
  • the antibody designated as “ATA009” comprises the V H CDR1, V H CDR2, and V H CDR3 sequences disclosed in Table 9; and the V L CDR1, V L CDR2, and V L CDR3 sequences disclosed in Table 9.
  • the ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155 (disclosed in Table 9) and a V L comprising the amino acid sequence set forth in SEQ ID NO: 156 (disclosed in Table 9).
  • NAI-1540294631v3 204 [00696]
  • the antibody designated as “ATA010” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 10; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 10.
  • the ATA001 antibody comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 170 (disclosed in Table 10) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 171 (disclosed in Table 10).
  • the antibody designated as “ATA011” comprises the V H CDR1, V H CDR2, and V H CDR3 sequences disclosed in Table 11; and the V L CDR1, V L CDR2, and V L CDR3 sequences disclosed in Table 11.
  • the ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183 (disclosed in Table 11) and a V L comprising the amino acid sequence set forth in SEQ ID NO: 184 (disclosed in Table 11).
  • the antibody designated as “ATA012” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 12; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 12
  • the ATA001 antibody comprises a V H comprising the amino acid sequence set forth in SEQ ID NO: 201 (disclosed in Table 12) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 202 (disclosed in Table 12).
  • the antibody designated as “ATA013” comprises the V H CDR1, V H CDR2, and V H CDR3 sequences disclosed in Table 13; and the V L CDR1, V L CDR2, and V L CDR3 sequences disclosed in Table 13.
  • the antibody designated as “ATA015” comprises the V H CDR1, V H CDR2, and V H CDR3 sequences disclosed in Table 15; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 15.
  • the ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246 (disclosed in Table 15) and a V L comprising the amino acid sequence set forth in SEQ ID NO: 247 (disclosed in Table 15).
  • the antibody designated as “ATA016” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 16; and the V L CDR1, V L CDR2, and V L CDR3 sequences disclosed in Table 16.
  • the ATA001 antibody comprises a V H comprising the amino acid sequence NAI-1540294631v3 205 set forth in SEQ ID NO: 262 (disclosed in Table 16) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 263 (disclosed in Table 16).
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a fragment crystallizable region (Fc region).
  • Fc region is the C-terminal region of an immunoglobulin heavy chain.
  • the Fc region can be a functional Fc region, a native Fc region, a recombinant Fc region, or a variant Fc region.
  • the Fc region comprises one or more heavy chain constant domains (e.g., CH2, and CH3).
  • the variant Fc region comprises one or more modifications (e.g., substitutions) in the CH2 domain.
  • the variant Fc region comprises one or more modifications (e.g., substitutions) in the CH3 domain.
  • the heavy chain constant region is chosen from IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE.
  • the heavy chain constant region is chosen from IgG1, IgG2, IgG3, and IgG4.
  • the heavy chain constant region is chosen from IgG1.
  • the heavy chain constant region is chosen from human IgG1.
  • the Fc region is an IgG1 Fc region.
  • the Fc region is a human IgG1 Fc region.
  • the Fc region is a native human IgG1 Fc region.
  • the Fc region comprises the amino acid sequence set forth in SEQ ID NO: 357. SEQ ID NO: 357 is provided below.
  • the Fc region comprises an amino acid sequence that is at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence set forth in SEQ ID NO: 357.
  • a “functional Fc region” possesses an “effector function” of a native sequence Fc region.
  • effector functions include C1q binding; complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors (e.g., B cell receptor; BCR), etc.
  • Such effector functions generally require the Fc region to be combined with a binding region or binding domain (e.g., an antibody variable region or domain) and can be assessed using various assays as disclosed.
  • a “native Fc region” encompasses a Fc region found in nature, and not manipulated, modified, and/or changed (e.g., isolated, purified, selected, including or combining with other sequences such as variable region sequences) by a human.
  • native human Fc regions include a native human IgG1 Fc region (non-A and A allotypes), a native human IgG2 Fc region, a native human IgG3 Fc region a native human IgG4 Fc region as well as naturally occurring variants of the foregoing.
  • a “variant Fc region” comprises an amino acid sequence which differs from that of a native sequence Fc region by virtue of at least one amino acid modification, (e.g., substituting, addition, or deletion) preferably one or more amino acid substitution(s).
  • the variant Fc region comprises at least one amino acid substitution compared to a native Fc region or to the Fc region of a parent polypeptide, e.g., from about 1 to about 10 amino acid substitutions, from about 1 to about 5 amino acid substitutions in a native sequence Fc region or in the Fc region of the parent polypeptide.
  • the variant Fc region described herein can possess at least about 80%, at least about 90%, at least about 95% or at least about 99% sequence identity with a native Fc region and/or with an Fc region of a parent polypeptide.
  • a variant Fc region may comprises a loss of effector function (e.g., silent Fc).
  • the constant region/framework region of the presently disclosed antibodies can be altered, for example, by amino acid substitution, to modify the properties of the antibody (e.g., to increase or decrease one or more of: antigen binding affinity, Fc receptor binding, antibody carbohydrate, for example, glycosylation, fucosylation, etc.
  • the presently disclosed anti-GPC3 antibody is a human antibody. In certain embodiments, the presently disclosed anti-GPC3 antibody is a humanized antibody. In certain embodiments, the presently disclosed anti-GPC3 antibody is a monoclonal antibody. In certain embodiments, the presently disclosed anti-GPC3 antibody is a human monoclonal antibody. In certain embodiments, the presently disclosed anti-GPC3 antibody is a humanized monoclonal antibody.
  • the presently disclosed antigen-binding fragment is a Fab, Fab’, F(ab’) 2 , Fv, or single chain variable fragment (scFv).
  • a presently disclosed antigen-binding fragment is an scFv.
  • the presently disclosed antigen-binding fragment is an scFv-Fc.
  • a scFv-Fc fragment comprises an scFv that is connected to a Fc region. NAI-1540294631v3 207 5.5.2.
  • Multispecific Molecules [00713] The presently disclosed subject matter further provides for multispecific molecules comprising an anti-GPC3 antibody or an antigen-binding portion thereof disclosed herein.
  • the multispecific molecules are multispecific antibodies.
  • the multispecific molecules e.g., multispecific antibodies
  • the multispecific molecules are bispecific molecules (e.g., bispecific antibodies).
  • the multispecific molecule binds to at least two different binding sites or target molecules.
  • the multispecific molecule comprises at least a first binding specificity for a GPC3 integrin, and a second binding specificity for a second target epitope.
  • the second target epitope can be a GPC3 epitope, or a non-GPC3 epitope, e.g., an epitope on a second target antigen different from GPC3.
  • the non-GPC3 epitope is the second target antigen is tumor antigen.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof can be derivatized or linked to more than one other functional molecule to generate multispecific molecules.
  • a presently disclosed anti-GPC3 antibody or an antigen-binding fragment thereof can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other binding molecules, such as another antibody, antibody fragment, peptide or binding mimetic, such that a multispecific molecule results.
  • the multispecific molecules of the presently disclosed subject matter can be prepared by conjugating the constituent binding specificities using methods known in the art. For example, each binding specificity of the bispecific molecule can be generated separately and then conjugated to one another. When the binding specificities are proteins or peptides, a variety of coupling or cross- linking agents can be used for covalent conjugation.
  • Non-limiting examples of cross-linking agents include protein A, carbodiimide, N-succinimidyl-S-acetyl-thioacetate (SAT A), 5, 5'-dithiobis(2- nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2- pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidom ethyl) cyclohaxane-l- carboxylate (sulfo-SMCC).
  • SAT A N-succinimidyl-S-acetyl-thioacetate
  • DTNB 5, 5'-dithiobis(2- nitrobenzoic acid)
  • oPDM o-phenylenedimaleimide
  • SPDP N-succinimidyl-3-(2- pyridyldithi
  • bispecific antibodies can be constructed, such tandem scFv molecules (taFv), diabodies (Db), or single chain diabodies (scDb), and fusion protein with human serum albumin (Ryutaro, et al., J Biol Chem 2011; 286: 1812-1818; Anja, et al., Blood 2000; 95(6): 2098- 2103; Weiner, et al., J. Immunology 1994; 152(5): 2385-2392; Dafne, et al., J Biol Chem 2007; 282: 12650-12660), but are devoid of Fc effector functions with distinct pharmacokinetic profiles.
  • BsIgG bispecific immunoglobulin G
  • IgG appended with an additional antigen-binding moiety iii
  • bispecific antibody fragments iv
  • bispecific fusion proteins iv
  • bispecific antibody conjugates iv
  • BslgG comprises heavy chains that are engineered for heterodimerization.
  • heavy chains can be engineered for heterodimerization using a “knobs-into-holes” strategy, a SEED platform, a common heavy chain (e.g., in ⁇ -bodies), and use of heterodimeric Fc regions.
  • Strategies are known in the art to avoid heavy chain pairing of homodimers in BsIgG, including knobs-into-holes, duobody, azymetric, charge pair, HA-TF, SEEDbody, and differential protein A affinity.
  • Another bispecific molecule format is IgG appended with an additional antigen-binding moiety.
  • bispecific antibody fragments are a format of bispecific molecules that lack some or all of the antibody constant domains. For example, some BsAb lack an Fc region.
  • bispecific antibody fragments include heavy and light chain regions that are connected by a peptide linker that permits efficient expression of the BsAb in a single host cell.
  • Bispecific fusion proteins include antibody fragments linked to other proteins.
  • bispecific fusion proteins can be linked to other proteins to add additional specificity and/or functionality.
  • the dock-and-lock (DNL) method can be used to generate bispecific antibody molecules with higher valency.
  • bispecific antibody fusions to albumin binding proteins or human serum albumin can be extend the serum half-life of antibody fragments.
  • chemical conjugation for example, chemical conjugation of antibodies and/or antibody fragments, can be used to create BsAb molecules.
  • NAI-1540294631v3 209 [00723] Methods of production of multispecific molecules, including bispecific molecules, are known in the art.
  • the presently disclosed subject matter further provides antibodies or antigen-binding fragments thereof that cross-compete for binding to a GPC3 antigen (e.g., a human GPC3 antigen) with any of the presently disclosed anti-GPC3 antibodies or antigen-binding fragments thereof (e.g., those disclosed in Sections 5.5.1 and 5.5.2, e.g., ATA001, ATA002, ATA003, ATA004, ATA005, ATA006, ATA007, ATA008, ATA009, ATA010, ATA011, ATA012, ATA013, ATA014, ATA015, and ATA016).
  • a GPC3 antigen e.g., a human GPC3 antigen
  • any of the presently disclosed anti-GPC3 antibodies or antigen-binding fragments thereof e.g., those disclosed in Sections 5.5.1 and 5.5.2, e.g., ATA001, ATA002, ATA003, ATA004, ATA005, ATA006,
  • the cross-competing antibodies bind to the same epitope region, e.g., same epitope, adjacent epitope, or overlapping as any of the presently disclosed anti-GPC4 antibodies or antigen-binding fragments thereof (e.g., those disclosed in Section 5.5.1 and 5.5.2, e.g., ATA001, ATA002, ATA003, ATA004, ATA005, ATA006, ATA007, ATA008, ATA009, ATA010, ATA011, ATA012, ATA013, ATA014, ATA015, and ATA016).
  • the same epitope region e.g., same epitope, adjacent epitope, or overlapping as any of the presently disclosed anti-GPC4 antibodies or antigen-binding fragments thereof (e.g., those disclosed in Section 5.5.1 and 5.5.2, e.g., ATA001, ATA002, ATA003, ATA004, ATA005, ATA006, ATA007, ATA008,
  • the competition can be determined by an assay in which the test antibody under study prevents or inhibits the specific binding of the reference antibody to a common epitope or a common antigen (e.g., a GPC3 antigen).
  • a common epitope or a common antigen e.g., a GPC3 antigen.
  • Numerous types of competitive binding assays can be used to determine if a test antibody competes with a reference antibody for binding to ⁇ 5 integrin (e.g., human GPC3).
  • assays examples include solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see, e.g., Stahli et al., (1983) Methods in Enzymology 9:242-253); solid phase direct biotin-avidin EIA (see, e.g., Kirkland et al., (1986) J.
  • RIA solid phase direct or indirect radioimmunoassay
  • EIA enzyme immunoassay
  • sandwich competition assay see, e.g., Stahli et al., (1983) Methods in Enzymology 9:242-253
  • solid phase direct biotin-avidin EIA see, e.g., Kirkland et al., (1986) J.
  • such an assay involves the use of a purified antigen (e.g., ⁇ 5 integrin, such as human ⁇ 5 integrin) bound to a solid surface or cells bearing either of an un-labelled test antigen binding protein (e.g., test ⁇ 5 integrin antibody) or a labeled reference antigen binding protein (e.g., reference anti- ⁇ 5 integrin antibody).
  • a purified antigen e.g., ⁇ 5 integrin, such as human ⁇ 5 integrin
  • an un-labelled test antigen binding protein e.g., test ⁇ 5 integrin antibody
  • a labeled reference antigen binding protein e.g., reference anti- ⁇ 5 integrin antibody
  • Competitive inhibition may be NAI-1540294631v3 210 measured by determining the amount of label bound to the solid surface or cells in the presence of the test antigen binding protein.
  • the test antigen binding protein is present in excess.
  • Antibodies identified by competition assay include antibodies binding to the same epitope as the reference antibody and/or antibodies binding to an adjacent epitope sufficiently proximal to the epitope bound by the reference for antibodies steric hindrance to occur (e.g., similar epitope or overlapping epitope).
  • a competing antibody when it is present in excess, it will inhibit specific binding of a reference antibody to a common epitope or common antigen by at least about 20%, for example, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96% or at least about 97%, at least about 98%, or at least about 99% or more. 5.5.4.
  • Immunoconjugates [00726] The presently disclosed subject further provides conjugates comprising a presently disclosed anti-GPC4 antibody or antigen-binding fragment thereof.
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof is conjugated to a therapeutic moiety, such as a cytotoxin, a drug (e.g., an immunosuppressant) or a radiotoxin.
  • a therapeutic moiety such as a cytotoxin, a drug (e.g., an immunosuppressant) or a radiotoxin.
  • Such conjugates are referred to herein as “immunoconjugates.”
  • Immunoconjugates that include one or more cytotoxins are referred to as “immunotoxins.”
  • a cytotoxin or cytotoxic agent includes any agent that is detrimental to (e.g., kills) cells.
  • Non-limiting examples include taxol (such as ricin, diphtheria, gelonin), cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof.
  • taxol such as ricin, diphtheria, gelonin
  • cytochalasin B such as ricin, diphtheria, gelonin
  • cytochalasin B such as ricin, diphtheria, gelonin
  • cytochalasin B such as ricin,
  • Therapeutic agents also include, for example, calecheamicin, aureastatin, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepa chlorambucil, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis- dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti
  • cytotoxins that can be conjugated to the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof include duocarmycins, NAI-1540294631v3 211 calicheamicins, maytansines and auristatins, and derivatives thereof.
  • a non-limiting example of a calicheamicin antibody conjugate is commercially available (MylotargTM; Wyeth-Ayerst).
  • Cytoxins can be conjugated to the presently disclosed anti-GPC3 antibody or antigen- binding fragment thereof using linker technology available in the art.
  • linker types that have been used to conjugate a cytotoxin to an antibody include, but are not limited to, hydrazones, thioethers, esters, disulfides and peptide-containing linkers.
  • a linker can be chosen that is, for example, susceptible to cleavage by low pH within the lysosomal compartment or susceptible to cleavage by proteases, such as proteases preferentially expressed in tumor tissue such as cathepsins (e.g., cathepsins B, C, D).
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof can also be conjugated to a radioactive isotope to generate cytotoxic radiopharmaceuticals, also referred to as radioimmunoconjugates.
  • radioactive isotopes that can be conjugated to antibodies for use diagnostically or therapeutically include, but are not limited to, Y, I, Ac, Bi, Ra, Lu, and Th. Method for preparing radioimmunoconjugates are established in the art.
  • radioimmunoconjugates are commercially available, including ZevalinTM (IDEC Pharmaceuticals) and BexxarTM (Corixa Pharmaceuticals), and similar methods can be used to prepare radioimmunoconjugates using the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof.
  • the conjugates can be used to modify a given biological response, and the drug moiety is not to be construed as limited to classical chemical therapeutic agents.
  • the drug moiety may be a protein or polypeptide possessing a desired biological activity.
  • Such proteins may include, for example, an enzymatically active toxin, or active fragment thereof, such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor (TNF) or interferon- ⁇ ; or, biological response modifiers such as, for example, lymphokines, interleukin-1 (“IL-1”), interleukin-2 (“IL-2”), interleukin-6 (“IL-6”), granulocyte macrophage colony stimulating factor (“GM-CSF”), granulocyte colony stimulating factor (“G-CSF”), or other growth factors.
  • TNF tumor necrosis factor
  • IL-6 granulocyte macrophage colony stimulating factor
  • G-CSF granulocyte colony stimulating factor
  • the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof can be prepared using an antibody or an antigen-binding fragment thereof comprising one or more of the V and/or V sequences disclosed herein as starting material to engineer a modified antibody, which modified antibody may have altered properties from the starting antibody.
  • An antibody can be engineered by modifying one or more residues within one or both variable regions (i.e., V and/or V), for example within one or more CDR regions and/or within one or more framework regions.
  • V and/or V variable regions
  • NAI-1540294631v3 212 Additionally or alternatively, an antibody can be engineered by modifying residues within the constant region(s), for example to alter the effector function(s) of the antibody.
  • CDR grafting One type of variable region engineering that can be performed is CDR grafting.
  • Antibodies interact with target antigens predominantly through amino acid residues that are located in the six heavy and light chain CDRs. For this reason, the amino acid sequences within CDRs are more diverse between individual antibodies than sequences outside of CDRs. Because CDR sequences are responsible for most antibody-antigen interactions, it is possible to express recombinant antibodies that mimic the properties of specific naturally occurring antibodies by constructing expression vectors that include CDR sequences from the specific naturally occurring antibody grafted onto framework sequences from a different antibody with different properties (see, e.g., Riechmann, L. et al. (1998) Nature 332:323-327; Jones, P. et al.
  • Framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences.
  • V CDRl, CDR2, and CDR3 sequences, and the V CDR1, CDR2, and CDR3 sequences can be grafted onto framework regions that have the identical sequence as that found in the germline immunoglobulin gene from which the framework sequence derive, or the CDR sequences can be grafted onto framework regions that contain one or more mutations as compared to the germline sequences.
  • Another type of variable region modification is to mutate amino acid residues within the V and/or V CDRl, CDR2 and/or CDR3 regions to thereby improve one or more binding properties (e.g., affinity) of the antibody of interest.
  • Site-directed mutagenesis or PCR-mediated mutagenesis can be performed to introduce the mutation(s) and the effect on antibody binding, or other functional property of interest, can be evaluated in in vitro or in vivo assays. In certain embodiments, conservative modifications are introduced.
  • the mutations may be amino acid substitutions, additions or deletions. For example, no more than one, two, three, four or five residues within a CDR region are altered.
  • anti-GPC3 antibodies or antigen-binding fragments thereof comprising a V comprising: (a) a CDR1 sequence of the antibodies and antigen-binding fragments thereof disclosed herein, or an amino acid sequence having at least one (e.g., no more than one, no more than two, no more than three, no more than four or no more than five) amino acid modification (e.g., substitution, deletion and/or addition) as compared to the V CDR1 sequence of any one of the antibodies or antigen-binding fragments NAI-1540294631v3 213 thereof disclosed herein; (b) a CDR2 sequence of any one of the antibodies or antigen-binding fragments thereof disclosed herein, or an amino acid sequence having at least one (e.g., no more than one, no more than two, no more than three, no more than four or no more than five) amino acid modification (e.g., substitution, deletion and/or addition) as compared to the V CDR2
  • Engineered antibodies of the presently disclosed subject matter include those in which modifications are made to framework residues within VH and/or VL, e.g., to improve the properties of the antibody. Typically such framework modifications are made to decrease the immunogenicity of the antibody.
  • one approach is to “backmutate” one or more framework residues to the corresponding germline sequence. More specifically, an antibody that has undergone somatic mutation may contain framework residues that differ from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the antibody framework sequences to the germline sequences from which the antibody is derived.
  • the presently disclosed anti-GPC3 antibodies or antigen-binding fragments thereof can be engineered NAI-1540294631v3 214 to include modifications within the Fc region, typically to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and/or antigen- dependent cellular cytotoxicity.
  • a presently disclosed anti-GPC3 antibody or antigen- binding fragment thereof can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or be modified to alter its glycosylation, again to alter one or more functional properties of the antibody.
  • the hinge region of CH1 can be modified such that the number of cysteine residues in the hinge region is altered, e.g., increased or decreased.
  • the number of cysteine residues in the hinge region of CH1 is altered to, for example, facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody.
  • the Fc hinge region of an antibody may be mutated to decrease the biological half life of the antibody. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of the Fc-hinge fragment such that the antibody has impaired Staphylococcyl protein A (SpA) binding relative to native Fc-hinge domain SpA binding.
  • SpA Staphylococcyl protein A
  • the antibody can be modified to increase its biological half-life, e.g., the antibody can be altered within the CH1 or CL region to contain a salvage receptor binding epitope taken from two loops of a CH2 domain of an Fc region of an IgG. Furthermore, the Fc region can be altered by replacing at least one amino acid residue with a different amino acid residue to alter the effector function(s) of the antibody.
  • the Fc region can be modified to increase the ability of the antibody to mediate antibody dependent cellular cytotoxicity (ADCC) and/or to increase the affinity of the antibody for an Fc ⁇ receptor.
  • a presently disclosed anti-GPC3 antibody comprises an afucosylated Fc region.
  • glycosylation of a presently disclosed anti-GPC3 antibody can be modified.
  • an aglycoslated antibody can be made (i.e., the antibody lacks glycosylation).
  • Glycosylation can be altered to, for example, increase the affinity of the antibody for antigen.
  • carbohydrate modifications can be accomplished by, for example, altering one or more sites of glycosylation within the antibody sequence.
  • one or more amino acid substitution can be made that result in elimination of one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site.
  • an antibody can be made that has an altered type of glycosylation, such as a hypofucosylated antibody having reduced amounts of fucosyl residues or an antibody having increased bisecting GlcNac structures. Such altered glycosylation patterns have been demonstrated to increase the ADCC ability of antibodies.
  • Such carbohydrate modifications can NAI-1540294631v3 215 be accomplished by, for example, expressing the antibody in a host cell with altered glycosylation machinery.
  • Another modification of the antibodies may be pegylation.
  • an antibody can be pegylated to, for example, increase the biological (e.g., serum) half-life of the antibody.
  • the antibody, or fragment thereof typically is reacted with polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups become attached to the antibody or antibody fragment.
  • PEG polyethylene glycol
  • the pegylation may be carried out via an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer).
  • polyethylene glycol is intended to encompass any of the forms of PEG that have been used to derivatize other proteins, such as mono (Ci-Ci) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol- maleimide.
  • the antibody to be pegylated may be an aglycosylated antibody.
  • Nucleic Acids and Vectors [00742] The presently disclosed subject matter provides nucleic acids encoding the presently disclosed binding molecules. In one aspect, the presently disclosed subject matter provides GPC3- targeted CARs (e.g., those disclosed in Section 5.3), and cells comprising such nucleic acids.
  • the nucleic acid further comprises a promoter that is operably linked to the nucleic acid encoding a presently disclosed GPC3-targeted CAR.
  • the presently disclosed subject matter provides nucleic acids encoding the anti-GPC3 antibodies or antigen-binding fragments thereof disclosed herein (e.g., those disclosed in Section 5.4) or a region thereof, and cells comprising such nucleic acids.
  • the nucleic acid comprises a first polynucleotide encoding an anti-GPC3 V H disclosed herein (e.g., one disclosed in Sections 5.3 and 5.4, e.g., one disclosed in Tables 1-16).
  • the first polynucleotide encodes an anti-GPC3 VH comprising the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262.
  • the first polynucleotide encodes an anti-GPC3 VH comprising a CDR1, a CDR2, and a CDR3 disclosed in Tables 1-16.
  • the nucleic acid comprises a second polynucleotide encoding an anti-GPC3 VL disclosed herein (e.g., one disclosed in Sections 5.3 and 5.4, e.g., one disclosed in Tables 1-16).
  • the second polynucleotide encodes an anti-GPC3 V L comprising the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, NAI-1540294631v3 216 SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263.
  • the second polynucleotide encodes an anti-GPC3 VL comprising a CDR1, a CDR2, and a CDR3 disclosed in Tables 1-16.
  • the nucleic acid further comprises a promoter that is operably linked to the nucleic acid encoding a presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof, or a region of a presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof (e.g., VH or VL).
  • the promoter is endogenous or exogenous.
  • the exogenous promoter is selected from the group consisting of an elongation factor (EF)-1 promoter, a cytomegalovirus immediate-early promoter (CMV) promoter, a simian virus 40 early promoter (SV40) promoter, a phosphoglycerate kinase (PGK) promoter, a metallothionein promoter, and Ubiquitin C promoter.
  • EF elongation factor
  • CMV cytomegalovirus immediate-early promoter
  • SV40 simian virus 40 early promoter
  • PGK phosphoglycerate kinase
  • Ubiquitin C promoter Ubiquitin C promoter.
  • the endogenous promoter is selected from a TCR alpha promoter, a TCR beta promoter, and a beta 2-microglobulin promoter.
  • the promoter is an inducible promoter.
  • the inducible promoter is selected from the group consisting of a NFAT transcriptional response element (TRE) promoter, a CD69 promoter, a CD25 promoter, an IL-2 promoter, a 4-1BB promoter, a PD1 promoter, and a LAG3 promoter.
  • TRE NFAT transcriptional response element
  • the presently disclosed subject matter also provides vectors comprising the presently disclosed nucleic acids.
  • the vector is an expression vector.
  • the nucleic acids can be delivered into cells by art-known methods or as described herein. Genetic modification of a cell can be accomplished by transducing a substantially homogeneous cell composition with a recombinant DNA construct.
  • a retroviral vector e.g., gammaretroviral vector or lentiviral vector
  • a presently disclosed nucleic acid can be cloned into a retroviral vector and expression can be driven from its endogenous promoter, from the retroviral long terminal repeat, or from a promoter specific for a target cell type of interest.
  • Expression of nucleic acids encoding CARs is typically achieved by operably linking a nucleic acid encoding the CAR polypeptide to a promoter and incorporating the construct into an expression vector.
  • Typical cloning vectors contain transcription and translation terminators, initiation sequences, and promoters useful for regulation of the expression of the desired nucleic acid sequence.
  • NAI-1540294631v3 217 The disclosed nucleic acid can be cloned into a number of types of vectors.
  • the nucleic acid can be cloned into a vector including, but not limited to a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid.
  • Vectors of particular interest include expression vectors, replication vectors, probe generation vectors, and sequencing vectors.
  • the expression vector may be provided to a cell in the form of a viral vector.
  • Viruses which are useful as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, and lentiviruses.
  • a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers.
  • the polynucleotide vectors are lentiviral or retroviral vectors.
  • a number of viral based systems have been developed for gene transfer into mammalian cells.
  • retroviruses provide a convenient platform for gene delivery systems.
  • a selected gene can be inserted into a vector and packaged in retroviral particles using techniques known in the art.
  • the recombinant virus can then be isolated and delivered to cells of the subject either in vivo or ex vivo.
  • a suitable promoter is the immediate early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strong constitutive promoter sequence capable of driving high levels of expression of any polynucleotide sequence operatively linked thereto.
  • EF-1 ⁇ Elongation Growth Factor-1 ⁇
  • constitutive promoter sequences may also be used, including, but not limited to the simian virus 40 (SV40) early promoter, MND (myeloproliferative sarcoma virus) promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) long terminal repeat (LTR) promoter, MoMuLV promoter, an avian leukemia virus promoter, an Epstein-Barr virus immediate early promoter, a Rous sarcoma virus promoter, as well as human gene promoters such as, but not limited to, the actin promoter, the myosin promoter, the hemoglobin promoter, and the creatine kinase promoter.
  • the promoter can alternatively be an inducible promoter.
  • inducible promoters include, but are not limited to a metallothionine promoter, a glucocorticoid promoter, a progesterone promoter, and a tetracycline promoter.
  • Additional promoter elements e.g., enhancers, regulate the frequency of transcriptional initiation. Typically, these are located in the region 30-110 bp upstream of the start site, although a NAI-1540294631v3 218 number of promoters have recently been shown to contain functional elements downstream of the start site as well. The spacing between promoter elements frequently is flexible, so that promoter function is preserved when elements are inverted or moved relative to one another.
  • the expression vector to be introduced into a cell can also contain either a selectable marker gene or a reporter gene or both to facilitate identification and selection of expressing cells from the population of cells sought to be transfected or infected through viral vectors.
  • the selectable marker may be carried on a separate piece of DNA and used in a co-transfection procedure. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers include, for example, antibiotic-resistance genes.
  • Reporter genes are used for identifying potentially transfected cells and for evaluating the functionality of regulatory sequences.
  • a reporter gene is a gene that is not present in or expressed by the recipient organism or tissue and that encodes a polypeptide whose expression is manifested by some easily detectable property, e.g., enzymatic activity. Expression of the reporter gene is assayed at a suitable time after the DNA has been introduced into the recipient cells.
  • Suitable reporter genes may include genes encoding luciferase, beta-galactosidase, chloramphenicol acetyl transferase, secreted alkaline phosphatase, or the green fluorescent protein gene.
  • Suitable expression systems are well known and may be prepared using known techniques or obtained commercially.
  • the construct with the minimal 5′ flanking region showing the highest level of expression of reporter gene is identified as the promoter.
  • Such promoter regions may be linked to a reporter gene and used to evaluate agents for the ability to modulate promoter-driven transcription.
  • Methods of introducing and expressing genes into a cell are known in the art.
  • the vector can be readily introduced into a host cell, e.g., mammalian, bacterial, yeast, or insect cell by any method in the art.
  • the expression vector can be transferred into a host cell by physical, chemical, or biological means.
  • Physical methods for introducing a polynucleotide into a host cell include calcium phosphate precipitation, lipofection, particle bombardment, microinjection, electroporation, and the like. Methods for producing cells comprising vectors and/or exogenous nucleic acids are well-known in the art. See, for example, Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York). NAI-1540294631v3 219 [00759] Biological methods for introducing a polynucleotide of interest into a host cell include the use of DNA and RNA vectors.
  • Viral vectors and especially retroviral vectors, have become the most widely used method for inserting genes into mammalian, e.g., human cells.
  • Chemical means for introducing a polynucleotide into a host cell include colloidal dispersion systems, such as macromolecule complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes.
  • An exemplary colloidal system for use as a delivery vehicle in vitro and in vivo is a liposome (e.g., an artificial membrane vesicle).
  • an exemplary delivery vehicle is a liposome.
  • the nucleic acid may be associated with a lipid.
  • the nucleic acid associated with a lipid may be encapsulated in the aqueous interior of a liposome, interspersed within the lipid bilayer of a liposome, attached to a liposome via a linking molecule that is associated with both the liposome and the oligonucleotide, entrapped in a liposome, complexed with a liposome, dispersed in a solution containing a lipid, mixed with a lipid, combined with a lipid, contained as a suspension in a lipid, contained or complexed with a micelle, or otherwise associated with a lipid.
  • Lipid, lipid/DNA or lipid/expression vector associated compositions are not limited to any particular structure in solution. For example, they may be present in a bilayer structure, as micelles, or with a “collapsed” structure. They may also simply be interspersed in a solution, possibly forming aggregates that are not uniform in size or shape.
  • Lipids are fatty substances which may be naturally occurring or synthetic lipids.
  • lipids include the fatty droplets that naturally occur in the cytoplasm as well as the class of compounds which contain long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, amino alcohols, and aldehydes. Lipids suitable for use can be obtained from commercial sources.
  • DMPC dimyristyl phosphatidylcholine
  • DCP dicetyl phosphate
  • Choi cholesterol
  • DMPG dimyristyl phosphatidylglycerol
  • Any targeted genome editing methods can also be used to deliver a presently disclosed GPC3-targeted CAR to a cell.
  • a CRISPR system is used to deliver a presently disclosed GPC3-targeted CAR.
  • zinc-finger nucleases are used to deliver a presently disclosed GPC3-targeted CAR.
  • a TALEN system is used to deliver a presently disclosed GPC3-targeted CAR.
  • NAI-1540294631v3 220 [00763] Clustered regularly-interspaced short palindromic repeats (CRISPR) system is a genome editing tool discovered in prokaryotic cells.
  • the system When utilized for genome editing, the system includes Cas9 (a protein able to modify DNA utilizing crRNA as its guide), CRISPR RNA (crRNA, contains the RNA used by Cas9 to guide it to the correct section of host DNA along with a region that binds to tracrRNA (generally in a hairpin loop form) forming an active complex with Cas9), trans- activating crRNA (tracrRNA, binds to crRNA and forms an active complex with Cas9), and an optional section of DNA repair template (DNA that guides the cellular repair process allowing insertion of a specific DNA sequence).
  • CRISPR/Cas9 often employs a plasmid to transfect the target cells.
  • the crRNA needs to be designed for each application as this is the sequence that Cas9 uses to identify and directly bind to the target DNA in a cell.
  • the repair template carrying CAR expression cassette need also be designed for each application, as it must overlap with the sequences on either side of the cut and code for the insertion sequence.
  • Multiple crRNA’s and the tracrRNA can be packaged together to form a single-guide RNA (sgRNA). This sgRNA can be joined together with the Cas9 gene and made into a plasmid in order to be transfected into cells.
  • ZFN zinc-finger nuclease
  • ZFN is an artificial restriction enzyme, which is generated by combining a zinc finger DNA-binding domain with a DNA-cleavage domain.
  • a zinc finger domain can be engineered to target specific DNA sequences which allows a zinc-finger nuclease to target desired sequences within genomes.
  • the DNA-binding domains of individual ZFNs typically contain a plurality of individual zinc finger repeats and can each recognize a plurality of base pairs.
  • the most common method to generate new zinc-finger domain is to combine smaller zinc-finger “modules” of known specificity.
  • the most common cleavage domain in ZFNs is the non-specific cleavage domain from the type IIs restriction endonuclease FokI.
  • HR homologous recombination
  • ZFNs can be used to insert the CAR expression cassette into genome.
  • Transcription activator-like effector nucleases are restriction enzymes that can be engineered to cut specific sequences of DNA TALEN system operates on almost the same principle as ZFNs. They are generated by combining a transcription activator-like effectors DNA- binding domain with a DNA cleavage domain.
  • Transcription activator-like effectors are composed of 33-34 amino acid repeating motifs with two variable positions that have a strong NAI-1540294631v3 221 recognition for specific nucleotides. By assembling arrays of these TALEs, the TALE DNA-binding domain can be engineered to bind desired DNA sequence, and thereby guide the nuclease to cut at specific locations in genome.
  • cDNA expression for use in polynucleotide therapy methods can be directed from any suitable promoter (e.g., the human cytomegalovirus (CMV), simian virus 40 (SV40), metallothionein promoters, or Ubiquitin C promoter), and regulated by any appropriate mammalian regulatory element or intron (e.g.
  • enhancers known to preferentially direct gene expression in specific cell types can be used to direct the expression of a nucleic acid.
  • the enhancers used can include, without limitation, those that are characterized as tissue- or cell-specific enhancers.
  • regulation can be mediated by the cognate regulatory sequences or, if desired, by regulatory sequences derived from a heterologous source, including any of the promoters or regulatory elements described above.
  • the components of a selected genome editing method are delivered as DNA constructs in one or more plasmids.
  • the components are delivered via viral vectors.
  • Common delivery methods include but is not limited to, electroporation, microinjection, gene gun, impalefection, hydrostatic pressure, continuous infusion, sonication, magnetofection, adeno-associated viruses, envelope protein pseudotyping of viral vectors, replication-competent vectors cis and trans-acting elements, herpes simplex virus, and chemical vehicles (e.g., oligonucleotides, lipoplexes, polymersomes, polyplexes, dendrimers, inorganic Nanoparticles, and cell-penetrating peptides).
  • the present disclosure provides methods for detecting GPC3 in a whole cell or tissue.
  • the method comprises: a) contacting a cell or tissue with an anti-GPC3 antibody or antigen-binding fragment thereof disclosed herein, wherein the anti-GPC3 antibody or antigen-binding fragment thereof comprises a detectable label; and b) determining the amount of the labeled anti-GPC3 antibody or antigen-binding fragment thereof bound to the cell or tissue.
  • the method is for detecting human GPC3.
  • b) determining the amount of the labeled anti-GPC3 antibody or antigen-binding fragment thereof bound to the cell or tissue comprises measuring the amount of detectable label associated with the cell or tissue, wherein the amount of bound antibody or antigen- binding fragment thereof indicates the amount of GPC3 (e.g., human GPC3) in the cell or tissue.
  • GPC3 e.g., human GPC3
  • the cell or tissue can be any cell or tissue, including any normal, healthy, abnormal, tumor, or cancer cells and tissues.
  • the present disclosure also provides methods for detecting or diagnosing an GPC3- associated disease, disorder, or condition.
  • the method comprises: (a) measuring an expression level of GPC3 in a cell or a tissue sample of a subject using an anti- GPC3 antibody or antigen-binding fragment thereof disclosed herein; and (b) comparing the expression level of GPC3 measured in (a) with a control expression level of GPC3, wherein an increase in the expression level of GPC3 measured in (a) as compared to the control expression level of GPC3 is indicative of an GPC3-associated disease, disorder, or condition.
  • control expression level of GPC3 is an expression level of GPC3 in a cell or a tissue sample of a subject not suffering from an GPC3-associated disease, disorder, or condition. 5.8.
  • Formulations and Administration [00771] In one aspect, the presently disclosed subject matter provides compositions comprising the presently disclosed cells comprising a presently disclosed GPC3-targeted CAR. In another aspect, the presently disclosed subject matter provides compositions comprising the anti-GPC3 antibodies or antigen-binding fragments thereof, the conjugates, or the multispecific molecules disclosed herein. [00772] In certain embodiments, the composition is a pharmaceutical composition that further comprises a pharmaceutically acceptable carrier.
  • compositions can be conveniently provided as sterile liquid preparations, e.g., isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which may be buffered to a selected pH.
  • sterile liquid preparations e.g., isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which may be buffered to a selected pH.
  • Liquid preparations are normally easier to prepare than gels, other viscous compositions, and solid compositions. Additionally, liquid compositions are somewhat more convenient to administer, especially by injection. Viscous compositions, on the other hand, can be formulated within the appropriate viscosity range to provide longer contact periods with specific tissues.
  • Liquid or viscous compositions can comprise carriers, which can be a solvent or dispersing medium containing, for example, water, saline, phosphate buffered saline, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like) and suitable mixtures thereof.
  • carriers can be a solvent or dispersing medium containing, for example, water, saline, phosphate buffered saline, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like) and suitable mixtures thereof.
  • NAI-1540294631v3 223 Compositions comprising the presently disclosed cells can be provided systemically or directly to a subject in need thereof.
  • the presently disclosed compositions are directly injected into an organ of interest.
  • the presently disclosed compositions are provided indirectly to the organ of interest, for example, by administration into the circulatory system (e.g., the tumor vasculature).
  • Expansion and differentiation agents can be provided prior to, during or after administration of compositions to increase production of cells in vitro or in vivo.
  • the quantity of cells comprising the GPC3-targeted CAR comprised in the presently disclosed compositions can vary for the subject being treated.
  • the presently disclosed composition comprises between about 1 ⁇ 10 4 and about 1 ⁇ 10 10 , between about 1 ⁇ 10 4 and about 1 ⁇ 10 7 , between about 1 ⁇ 10 5 and about 1 ⁇ 10 7 , between about 1 ⁇ 10 5 and about 1 ⁇ 10 8 , between about 1 ⁇ 10 5 and about 10 9 , between about 1 ⁇ 10 6 and about 1 ⁇ 10 8 , between about 1 ⁇ 10 6 and about 1 ⁇ 10 9 , or between about 1 ⁇ 10 6 and about 1 ⁇ 10 10 of the presently disclosed cells comprising the GPC3-targeted CAR.
  • the presently disclosed composition comprises between about 1 ⁇ 10 5 and about 5 ⁇ 10 8 of the presently disclosed cells comprising the GPC3-targeted CAR.
  • the presently disclosed composition comprises between about 1 ⁇ 10 5 and about 1 ⁇ 10 7 of the presently disclosed cells comprising the GPC3-targeted CAR. In certain embodiments, the presently disclosed composition comprises between about 1 ⁇ 10 6 and about 1 ⁇ 10 8 of the presently disclosed cells comprising the GPC3-targeted CAR. In certain embodiments, the presently disclosed composition comprises between about 1 ⁇ 10 6 and about 5 ⁇ 10 8 of the presently disclosed cells comprising the GPC3-targeted CAR. More effective cells may be administered in even smaller numbers. Usually, at least about 1 ⁇ 10 5 cells will be administered, eventually reaching about 1 ⁇ 10 10 or more.
  • the presently disclosed composition comprises at least about 1 ⁇ 10 5 , 5 ⁇ 10 5 , 1 ⁇ 10 6 , about 5 ⁇ 10 6 , about 1 ⁇ 10 7 , about 2.5 ⁇ 10 7 , about 5 ⁇ 10 7 , about 1 ⁇ 10 8 , about 1.5 ⁇ 10 8 , about 2 ⁇ 10 8 , or about 5 ⁇ 10 8 of the presently disclosed cells comprising the GPC3-targeted CAR.
  • the presently disclosed composition comprises about 1 ⁇ 10 6 of the presently disclosed cells comprising the GPC3-targeted CAR. The precise determination of what would be considered an effective dose can be based on factors individual to each subject, including their size, age, sex, weight, and condition of the particular subject.
  • compositions can be administered multiple times at these dosages.
  • the presently disclosed CAR-modified cells may be administered either alone, or as a pharmaceutical composition in combination with diluents and/or with other components such as IL- 2, IL-15, or other cytokines or cell populations.
  • pharmaceutical compositions may comprise NAI-1540294631v3 224 a target cell population as described herein, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients.
  • compositions may comprise buffers such as neutral buffered saline, phosphate buffered saline and the like; carbohydrates such as glucose, mannose, sucrose or dextrans, mannitol; proteins; polypeptides or amino acids such as glycine; antioxidants; chelating agents such as EDTA or glutathione; adjuvants (e.g., aluminum hydroxide); and preservatives.
  • buffers such as neutral buffered saline, phosphate buffered saline and the like
  • carbohydrates such as glucose, mannose, sucrose or dextrans, mannitol
  • proteins polypeptides or amino acids such as glycine
  • antioxidants e.g., chelating agents such as EDTA or glutathione
  • adjuvants e.g., aluminum hydroxide
  • preservatives e.g., aluminum hydroxide
  • compositions may be carried out in any convenient manner, including by injection, transfusion, or implantation.
  • the presently disclosed compositions can be administered by any method known in the art including, but not limited to, intravenous administration, intradermal administration, subcutaneous administration, intranodal administration, intratumoral administration, intramedullary administration, intramuscular administration, intrathecal administration, intrapleural administration, intraosseous administration, intraperitoneal administration, pleural administration, and direct administration to the subject.
  • the composition is administered to a subject intravenously.
  • the compositions can be injected directly into a tumor, lymph node, or site of disease. 5.9.
  • the presently disclosed GPC3-targeted CAR-expressing cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used for treating a GPC3-associated disease, disorder, or condition, including one or more symptoms of the disease, disorder, or condition.
  • the presently disclosed subject matter provides methods of treating a GPC3-associated disease, disorder, or condition.
  • the method comprises administering to a subject in need thereof a presently disclosed anti-GPC3 antibody, antigen-binding fragment thereof, multispecific molecule, conjugate, or composition.
  • the method comprises administering to a subject in need thereof a composition comprising cells comprising a presently disclosed GPC3-targeted CAR.
  • the subject suffers from or is diagnosed with a GPC3-associated disease, disorder, or condition.
  • the subject is human.
  • NAI-1540294631v3 225 [00779] Immune effector cells expressing the presently disclosed GPC3-targeted CARs can elicit an immune response against GPC3-expressing cells.
  • the immune response elicited by the disclosed CAR-modified immune effector cells may be an active or a passive immune response.
  • the CAR-mediated immune response may be part of an adoptive immunotherapy approach in which CAR-modified immune effector cells induce an immune response specific to GPC3.
  • adoptive immunotherapy approach in which CAR-modified immune effector cells induce an immune response specific to GPC3.
  • Adoptive transfer of immune effector cells expressing CARs is a promising anti-cancer therapeutic. Following the collection of a subject’s immune effector cells, the cells may be genetically engineered to express the presently disclosed GPC3-targeted CARs, then infused back into the subject. Moreover, cells (e.g., immune effector cells) obtained from a donor other than the subject (e.g., allogeneic to the subject) may be genetically engineered to express the disclosed GPC3-targeted CARs, then the CAR-expressing cells infused into the subject.
  • the cells comprising a GPC3-targeted CAR polypeptide are allogeneic EBV-specific cytotoxic T cells.
  • the presently disclosed subject matter provides various methods of using the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions.
  • the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions can be used in a therapy or medicament.
  • the presently disclosed subject matter provides methods for inducing and/or increasing an immune response in a subject in need thereof.
  • the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions can be used for treating a GPC3-associated disease or disorder in a subject.
  • the GPC3-associated disease or disorder in a subject is a tumor.
  • the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions can be used for reducing tumor burden in a subject, and/or prolonging the survival of a subject suffering from a tumor.
  • the presently disclosed cells and compositions comprising thereof can reduce the number of tumor cells, reduce tumor size, and/or eradicate the tumor in the subject.
  • each of the above-noted method comprises administering the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions to achieve the desired effect, e.g., palliation of an existing condition or prevention of recurrence.
  • the amount administered is an amount effective in producing the desired effect.
  • An effective amount can be provided in one or a series of administrations.
  • An effective amount can be provided in a bolus or by continuous perfusion.
  • NAI-1540294631v3 226 [00782]
  • the GPC3-associated disease or disorder overexpresses GPC3.
  • Non-limiting examples of GPC3-associated disease or disorder include hepatocellular carcinoma, hepatoblastoma, lung squamous cell carcinoma, ovarian yolk sac tumor, melanoma, and urothelial carcinoma.
  • the GPC3-associated disease or disorder is a tumor.
  • the GPC3-associated disease or disorder is cancer.
  • the GPC3-associated disease or disorder is hepatocellular carcinoma.
  • the subject is a human subject. The subjects can have an advanced form of disease, in which case the treatment objective can include mitigation or reversal of disease progression, and/or amelioration of side effects.
  • the subjects can have a history of the condition, for which they have already been treated, in which case the therapeutic objective will typically include a decrease or delay in the risk of recurrence.
  • the therapeutic objective will typically include a decrease or delay in the risk of recurrence.
  • the presently disclosed cells, anti-GPC3 antibodies, antigen- binding fragments thereof, multispecific molecules, conjugates and compositions are administered to a subject in conjunction with (e.g., before, simultaneously or following) any number of relevant treatment modalities.
  • the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used in combination with chemotherapy, radiation, immunosuppressive agents, immunoablative agents, cytoxin, and cytokines.
  • the GPC3-associated disease or disorder is cancer.
  • the cancer can be any neoplasm or tumor for which radiotherapy is currently used.
  • the cancer can be a neoplasm or tumor that is not sufficiently sensitive to radiotherapy using standard methods.
  • a representative but non-limiting list of cancers that the disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used to treat include hepatocellular carcinoma, hepatoblastoma, lung squamous cell carcinoma, ovarian yolk sac tumor, melanoma, and urothelial carcinoma.
  • the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used in combination with any compound, moiety or group which has a cytotoxic or cytostatic effect.
  • Drug moieties include NAI-1540294631v3 227 chemotherapeutic agents, which may function as microtubulin inhibitors, mitosis inhibitors, topoisomerase inhibitors, or DNA intercalators, and particularly those which are used for cancer therapy.
  • chemotherapeutic agents which may function as microtubulin inhibitors, mitosis inhibitors, topoisomerase inhibitors, or DNA intercalators, and particularly those which are used for cancer therapy.
  • the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used in combination with a checkpoint inhibitor.
  • the two known inhibitory checkpoint pathways involve signaling through the cytotoxic T- lymphocyte antigen-4 (CTLA-4) and programmed-death 1 (PD-1) receptors.
  • CTLA-4 cytotoxic T- lymphocyte antigen-4
  • PD-1 programmed-death 1
  • the PD-1 receptor (also known as CD279) is expressed on the surface of activated T cells. Its ligands, PD-L1 (B7-H1; CD274) and PD-L2 (B7-DC; CD273), are expressed on the surface of APCs such as dendritic cells or macrophages. PD-L1 is the predominant ligand, while PD-L2 has a much more restricted expression pattern. When the ligands bind to PD-1, an inhibitory signal is transmitted into the T cell, which reduces cytokine production and suppresses T- cell proliferation.
  • Checkpoint inhibitors include, but are not limited to antibodies that block PD-1 (e.g., Nivolumab (BMS-936558 or MDX1106), CT-011, MK-3475), PD-L1 (e.g., MDX-1105 (BMS-936559), MPDL3280A, MSB0010718C), PD-L2 (e.g., rHIgM12B7), CTLA-4 (e.g., Ipilimumab (MDX-010), Tremelimumab (e.g., CP-675,206)), IDO, B7-H3 (e.g., MGA271), B7-H4, TIM3, LAG-3 (e.g., BMS-986016).
  • PD-1 e.g., Nivolumab (BMS-936558 or MDX1106), CT-011, MK-3475
  • PD-L1 e.g., MDX-1105 (BMS-936559), MPDL3280A
  • T cell receptor activation plus co-stimulation can help generate optimal “killer” CD8 T cell responses, which can be provided through ligation of tumor necrosis factor receptor family members, including OX40 (CD134) and 4-1BB (CD137).
  • OX40 is of particular interest as treatment with an activating (agonist) anti-OX40 mAb augments T cell differentiation and cytolytic function leading to enhanced anti-tumor immunity against a variety of tumors.
  • the presently disclosed method comprises administering to the subject a second therapeutic agent.
  • the second therapeutic agent is an antimetabolite, such as methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, fludarabine, 5- fluorouracil, decarbazine, hydroxyurea, asparaginase, gemcitabine or cladribine.
  • an antimetabolite such as methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, fludarabine, 5- fluorouracil, decarbazine, hydroxyurea, asparaginase, gemcitabine or cladribine.
  • the second therapeutic agent is an alkylating agent, such as mechlorethamine, thioepa, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, dacarbazine (DTIC), procarbazine, mitomycin C, cisplatin and other platinum derivatives, such as carboplatin.
  • alkylating agent such as mechlorethamine, thioepa, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, dacarbazine (DTIC), procarbazine, mitomycin C, cisplatin and other platinum derivatives, such as carboplatin.
  • the second therapeutic agent is an anti-mitotic agent, such as taxanes, for instance docetaxel, and paclitaxel, and vinca alkaloids, for instance vindesine, vincristine, vinblastine, and vinorelbine.
  • the second therapeutic agent is a topoisomerase inhibitor, such as topotecan or irinotecan, or a cytostatic drug, such as etoposide and teniposide.
  • the second therapeutic agent is a growth factor inhibitor, such as an inhibitor of ErbBl (EGFR) (such as an EGFR antibody, e.g.
  • EGFR ErbBl
  • the second therapeutic agent is a tyrosine kinase inhibitor, such as imatinib (Glivec, Gleevec STI571) or lapatinib.
  • the second therapeutic agent is a cytokine, growth factor, chemokine, or a combination thereof.
  • cytokines and growth factors include IFN ⁇ , IL-2, IL-4, IL-6, IL-7, IL-10, IL-12, IL-13, IL-15, IL-18, IL-23, IL-24, IL-27, IL-28a, IL-28b, IL-29, KGF, IFN ⁇ (e.g., INF ⁇ 2b), IFN , GM-CSF, CD40L, Flt3 ligand, stem cell factor, ancestim, and TNF ⁇ .
  • Non-limiting examples of chemokines include Glu-Leu-Arg (ELR)-negative chemokines such as IP-10, MCP-3, MIG, and SDF-la from the human CXC and C-C chemokine families.
  • Suitable cytokines include cytokine derivatives, cytokine variants, cytokine fragments, and cytokine fusion proteins.
  • the second therapeutic agent is a cell cycle control/apoptosis regulator (or “regulating agent”).
  • a cell cycle control/apoptosis regulator may include molecules that target and modulate cell cycle control/apoptosis regulators such as (i) cdc-25 (such as NSC 663284), (ii) cyclin-dependent kinases that overstimulate the cell cycle (such as flavopiridol (L868275, HMR1275), 7-hydroxystaurosporine (UCN-01, KW-2401), and roscovitine (R- roscovitine, CYC202)), and (iii) telomerase modulators (such as BIBR1532, SOT-095, GRN163 and compositions described in for instance US 6,440,735 and US 6,713,055) .
  • cdc-25 such as NSC 663284
  • cyclin-dependent kinases that overstimulate the cell cycle such as flavopiridol (L868275, HMR1275), 7-hydroxystaurosporine (UCN-01, KW-2401), and
  • Non-limiting examples of molecules that interfere with apoptotic pathways include TNF-related apoptosis-inducing ligand (TRAIL)/apoptosis-2 ligand (Apo-2L), antibodies that activate TRAIL receptors, IFNs, and anti- sense Bcl-2.
  • TRAIL TNF-related apoptosis-inducing ligand
  • Apo-2L apoptosis-2 ligand
  • antibodies that activate TRAIL receptors IFNs
  • anti- sense Bcl-2 anti- sense Bcl-2.
  • the second therapeutic agent is a hormonal regulating agent, such as agents useful for anti-androgen and anti-estrogen therapy.
  • hormonal regulating agents examples include tamoxifen, idoxifene, fulvestrant, droloxifene, toremifene, raloxifene, NAI-1540294631v3 229 diethylstilbestrol, ethinyl estradiol/estinyl, an antiandrogene (such as flutaminde/eulexin), a progestin (such as such as hydroxyprogesterone caproate, medroxy- progesterone/provera, megestrol acepate/megace), an adrenocorticosteroid (such as hydrocortisone, prednisone), luteinizing hormone- releasing hormone (and analogs thereof and other LHRH agonists such as buserelin and goserelin), an aromatase inhibitor (such as anastrazole/arimidex, aminoglutethimide/cytraden, exemestane) or a hormone
  • the second therapeutic agent is an anti-cancer nucleic acid or an anti-cancer inhibitory RNA molecule.
  • Combined administration as described above, may be simultaneous, separate, or sequential.
  • the agents may be administered as one composition or as separate compositions, as appropriate.
  • the presently disclosed cells, anti-GPC3 antibodies, antigen- binding fragments thereof, multispecific molecules, conjugates and compositions are administered in combination with radiotherapy.
  • Radiotherapy may comprise radiation or associated administration of radiopharmaceuticals to a subject is provided.
  • the source of radiation may be either external or internal to the patient being treated (radiation treatment may, for example, be in the form of external beam radiation therapy (EBRT) or brachytherapy (BT)).
  • Radioactive elements that may be used in practicing such methods include, e.g., radium, cesium-137, iridium-192, americium-241, gold-198, cobalt-57, copper-67, technetium-99, iodide-123, iodide-131, and indium-111.
  • radium cesium-137
  • iridium-192 e.g., americium-241, gold-198, cobalt-57, copper-67, technetium-99, iodide-123, iodide-131, and indium-111.
  • the presently disclosed cells, anti-GPC3 antibodies, antigen- binding fragments thereof, multispecific molecules, conjugates and compositions are administered in combination with surgery.
  • the GPC3-targeted CAR-T cells may be designed in several ways that enhance tumor cytotoxicity and specificity, evade tumor immunosuppression, avoid host rejection, and prolong their therapeutic half-life.
  • TRUCK T-cells Redirected for Universal Cytokine Killing
  • T cells possess a CAR but are also engineered to release cytokines such as IL-12 that promote tumor killing. Because these cells are designed to release a molecular payload upon activation of the CAR once localized to the tumor environment, these CAR-T cells are sometimes also referred to as ‘armored CARs’.
  • cytokines as cancer therapies are being investigated both pre-clinically and clinically, and may also prove useful when similarly incorporated into a TRUCK form of CAR- T therapy.
  • these include IL-2, IL-3.
  • “Self-driving” or “homing” CAR-T cells are engineered to express a chemokine NAI-1540294631v3 230 receptor in addition to their CAR. As certain chemokines can be upregulated in tumors, incorporation of a chemokine receptor aids in tumor trafficking to and infiltration by the adoptive T- cell, thereby enhancing both specificity and functionality of the CAR-T (Moon 2011). Universal CAR-T cells also possess a CAR, but are engineered such that they do not express endogenous TCR (T-cell receptor) or MHC (major histocompatibility complex) proteins.
  • T-cell receptor T-cell receptor
  • MHC major histocompatibility complex
  • Armored CAR-T cells are additionally so named for their ability to evade tumor immunosuppression and tumor-induced CAR-T hypofunction.
  • These particular CAR-Ts possess a CAR, and may be engineered to not express checkpoint inhibitors.
  • these CAR-Ts can be co-administered with a monoclonal antibody (mAb) that blocks checkpoint signaling.
  • mAb monoclonal antibody
  • Administration of an anti-PD-L1 antibody significantly restored the killing ability of CAR TILs (tumor infiltrating lymphocytes).
  • TILs include LAG-3, Tim-3, IDO-1, 2B4, and KIR.
  • Other intracellular inhibitors of TILs include phosphatases (SHP1), ubiquitin-ligases (i.e., cbl-b), and kinases (i.e., diacylglycerol kinase) .
  • Armored CAR-Ts may also be engineered to express proteins or receptors that protect them against or make them resistant to the effects of tumor-secreted cytokines.
  • Tandem and dual CAR-T cells are unique in that they possess two distinct antigen binding domains.
  • a tandem CAR contains two sequential antigen binding domains facing the extracellular environment connected to the intracellular costimulatory and stimulatory domains.
  • a dual CAR is engineered such that one extracellular antigen binding domain is connected to the intracellular costimulatory domain and a second, distinct extracellular antigen binding domain is connected to the intracellular stimulatory domain.
  • dual CARs are also referred to as “split CARs”. In both tandem and dual CAR designs, binding of both antigen binding domains is necessary to allow signaling of the CAR circuit in the T-cell. Because these two CAR designs have binding affinities for different, distinct antigens, they are also referred to as “bi-specific” CARs. [00805] To better control CAR-T therapy and prevent against unwanted side effects, a variety of features have been engineered including off-switches, safety mechanisms, and conditional control NAI-1540294631v3 231 mechanisms.
  • Both self-destruct and marked/tagged CAR-T cells are engineered to have an “off-switch” that promotes clearance of the CAR-expressing T-cell.
  • a self-destruct CAR-T contains a CAR, but is also engineered to express a pro-apoptotic suicide gene or “elimination gene” inducible upon administration of an exogenous molecule.
  • suicide genes may be employed for this purpose, including HSV-TK (herpes simplex virus thymidine kinase), Fas, iCasp9 (inducible caspase 9), CD20, MYC TAG, and truncated epidermal growth factor receptor (EGFRt) polypeptide (endothelial growth factor receptor).
  • HSK for example, converts the prodrug ganciclovir (GCV) into GCV-triphosphate that incorporates itself into replicating DNA, ultimately leading to cell death.
  • iCasp9 is a chimeric protein containing components of FK506-binding protein that binds the small molecule AP1903, leading to caspase 9 dimerization and apoptosis.
  • the suicide gene is an EGFRt polypeptide.
  • the EGFRt polypeptide can enable T-cell elimination by administering anti-EGFR monoclonal antibody (e.g., cetuximab).
  • EGFRt can be covalently joined to the upstream of the GPC3-targeted CAR.
  • the suicide gene can be included within the vector comprising nucleic acids encoding a presently disclosed GPC3-targeted CAR.
  • a prodrug designed to activate the suicide gene e.g., a prodrug (e.g., AP1903 that can activate iCasp-9) during malignant T-cell transformation (e.g., GVHD) triggers apoptosis in the suicide gene-activated cells expressing the GPC3-targeted CAR.
  • a prodrug designed to activate the suicide gene e.g., a prodrug (e.g., AP1903 that can activate iCasp-9) during malignant T-cell transformation (e.g., GVHD) triggers apoptosis in the suicide gene-activated cells expressing the GPC3-targeted CAR.
  • a prodrug designed to activate the suicide gene e.g., a prodrug (e.g., AP1903 that can activate i
  • a presently disclosed cell incorporated with a suicide gene can be pre-emptively eliminated at a given timepoint post the cell infusion, or eradicated at the earliest signs of toxicity.
  • a marked/ tagged CAR-T cell is one that possesses a CAR but also is engineered to express a selection marker. Administration of a mAb against this selection marker may promote clearance of the CAR-T cell.
  • Truncated EGFR is one such targetable antigen by the anti-EGFR mAb, and administration of cetuximab works to promotes elimination of the CAR-T cell.
  • CARs created to have these features are also referred to as sCARs for “switchable CARs”, and RCARs for ‘regulatable CARs’.
  • a “safety CAR”, also known as an “inhibitory CAR” (iCAR) is engineered to express two antigen binding domains. One of these extracellular domains is directed against a tumor related antigen and bound to an intracellular costimulatory and stimulatory domain. The second extracellular antigen binding domain however is specific for normal tissue and bound to an intracellular checkpoint domain such as CTLA-4, PD-1, or CD45. Incorporation of multiple intracellular inhibitory domains to the iCAR is also possible.
  • CEACAM CEACAM-1. NAI-1540294631v3 232 CEACAM-3, and/or CEACAM-5
  • LAG-3 TIGIT
  • BTLA BTLA
  • LAIR1 TGF ⁇ -R
  • the safety CAR-T engineering enhances specificity of the CAR-T cell for tumor tissue, and is advantageous in situations where certain normal tissues may express very low levels of a tumor associated antigen that would lead to off target effects with a standard CAR (Morgan 2010).
  • a conditional CAR-T cell expresses an extracellular antigen binding domain connected to an intracellular costimulatory domain and a separate, intracellular costimulator.
  • the costimulatory and stimulatory domain sequences are engineered in such a way that upon administration of an exogenous molecule the resultant proteins will come together intracellularly to complete the CAR circuit. In this way, CAR-T activation can be modulated, and possibly even “fine-tuned” or personalized to a specific patient.
  • two or more of these engineered features may be combined to create an enhanced, multifunctional CAR-T.
  • a CAR-T cell with either dual- or conditional-CAR design that also releases cytokines like a TRUCK.
  • a dual-conditional CAR-T cell could be made such that it expresses two CARs with two separate antigen binding domains against two distinct cancer antigens, each bound to their respective costimulatory domains.
  • Example 1 Generation of GPC3-Targeted scFv-Fc Fusion Proteins NAI-1540294631v3 233
  • a total of 16 human binders for human GPC3 were obtained as described in Section 5.3.1. To characterize the binders, 16 binders were formatted as fusion proteins with IgG1 (Kappa) Fc domain and produced.
  • VH and VL of a binder were coupled through a linker (e.g., a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1) to form a single chain variable region (scFv).
  • the scFv was further fused with a C-terminal Fc domain (e.g., a Fc domain comprising the amino acid sequence set forth in SEQ ID NO: 357) in order to purify the expressed scFv.
  • the fusion protein was designated as “scFv-Fc.”
  • the 16 scFv-Fc fusion proteins were designated as “ATA001 VH-VK Fc” (or “1-VH-linker-VL” shown in Figures 1A-1E), “ATA002 VH-VK Fc” (or “2-VH- linker-VL” shown in Figures 1A-1E), “ATA003 VH-VK Fc” (or “3-VH-linker-VL” shown in Figures 1A-1E), “ATA004 VH-VK Fc” (or “4-VH-linker-VL” shown in Figures 1A-1E), “ATA005 VH-VK Fc” (or “5-VH-linker-VL” shown in Figures 1A-1E), “ATA006 VH-VK Fc” (or “6-VH- linker-VL” shown in Figures 1A-1E), “ATA007 VH-VK Fc”
  • scFv-Fc of 16 binders To facilitate expression, codon optimization for scFv-Fc was performed as described in Section 5.3.1. [00812] To produce scFv-Fc of 16 binders, plasmids encoding scFv-Fc were introduced into Expi293F cells or ExpiCHO-S cells by transient transfection using standard procedures. The scFv- Fc of 16 binders were purified from culture supernatant produced using standard procedures of Protein A magnetic bead affinity purification. The scFv-Fc fusion proteins for 16 binders were expressed and purified with a purity of greater than 90%. Example 2.
  • binding activities of scFv-Fc Fusion Proteins [00813] The binding activities of the scFv-Fc fusion proteins described in Example 1 to human GPC3 were assessed by using enzyme-linked immunosorbent assay (ELISA) and fluorescence- activated cell sorting (FACS). [00814] As shown in the ELISA results of Table 17, all tested scFv-Fc fusion proteins showed specific binding to GPC36 ⁇ His protein. In addition, the FACS results demonstrate the specific binding of all tested scFv-Fc fusion proteins to human GPC3.
  • ELISA enzyme-linked immunosorbent assay
  • FACS fluorescence- activated cell sorting
  • GC33 antibody against GPC3 was used as benchmark.
  • Competitive binding activity was used to cluster the binders by epitope bins.
  • Final concentration of each biotinylated scFv-Fc protein designated as “-Bio” used for competition ELISA were shown in Table 18. Self-competition was highlighted. Results indicated that the binders exhibited various levels of competition. Based on the competitive and non- competitive relationship, the binders were grouped by epitope bins. The 16 binders were shown to bind to five distinct epitopes: Epitope 1, Epitope 2, Epitope 3, Epitope 4, and Epitope 5.
  • ATA001, ATA002, ATA003, ATA004, ATA005, ATA006, and ATA011 can be binned together and in the same group as GC33 antibody that bound to Epitope 1.
  • ATA007, ATA009, and NAI-1540294631v3 235 ATA015 were shown to bind to Epitope 2.
  • ATA008, ATA009, ATA010, and ATA013 were shown to bind to Epitope 3; ATA014 and ATA016 were shown to bind to Epitope 4; and ATA012 was shown to bind to Epitope 5.
  • Epitope 1 is within amino acids 524-563 of human GPC3 (e.g., one comprising the amino acid sequence set forth in SEQ ID NO: 22). Epitope 3 is at the C-terminus of GPC3. NAI-1540294631v3 236 - 6 1 c 0 F l m AK o i / 1 0 0 7 2 1 9 3 4 9 0 7 7 5 1 4 1 7 7 6 8 5 5 3 3 9 T V - Bg n 0 3 . 1 4 . 1 2 . 1 3 . 1 . 1 . 1 . 1 . 8 . 1 . 4 . 0 . AH .
  • GPC3-targeted CAR T Cells Twelve (12) GPC3-targeted CARs were constructed.
  • six (6) GPC3 binders ATA001, ATA002, ATA004, ATA008, ATA009, and ATA016 in both V H -V L orientation and VL-VH orientation were used to generate SFG 28z1XX P2A LNGFR CAR constructs.
  • FIG. 2A The general structure of the CAR is illustrated in Figure 2A.
  • the scFv was preceded by a leader peptide and followed by CD28 hinge-transmembrane-intracellular regions, and ITAM-mutated CD3 ⁇ intracellular domains (“1XX”) linked to a P2A sequence to induce co-expression of truncated low-affinity nerve growth factor receptor (LNGFR).
  • LNGFR truncated low-affinity nerve growth factor receptor
  • T cells were transduced with retroviral supernatants by centrifugation. Transduction efficiencies were determined 4 days later by flow cytometry.
  • the flow cytometric analysis shows the expression levels of LNGFR for the indicated constructs in the VH-VK and VK-VH orientation.
  • Untransduced T (UT) cells were used as a negative control.
  • the results indicate that the twelve (12) GPC3-targeted CARs were successfully transduced into and expressed by the host T cells and showed comparable levels of transduction efficiency.
  • the T cell exhaustion of the twelve (12) GPC3-targeted CARs was assessed by CD3/CD28 beads activation. On day 8 post activation, a fluorescence-activated cell sorting (FACS) analysis was performed to analyze the exhaustion markers LAG3, PD1, and TIM3 on the CAR T cells. Analysis shows no difference in expression of LAG3, PD1 or TIM3 between VH-VK and VK-VH orientation CAR T cells (data not shown).
  • FACS fluorescence-activated cell sorting
  • the GPC3 expression levels of Nalm-6 GPC3 + , Huh7, and PLC/PRF/5 tumor cells are shown in Figure 3.
  • Tumor cell killing was measured in a bioluminescence (BLI) assay at the indicated effector:target (E:T) ratios.
  • the effector and tumor target cells were co-cultured in triplicates at the indicated effector/target ratio using black-walled 96-well plates with 1 ⁇ 10 4 target cells in a total volume of 100 ⁇ per well in medium.
  • Target cells alone were plated at the same cell density to determine the maximal luciferase expression (relative light units (RLU); 18 hours later, 100 ⁇ L luciferase substrate was directly added to each well. Emitted light was detected in a luminescence plate reader. Lysis was determined as (1 – (RLUsample)/(RLUmax)) ⁇ 100.
  • RLU relative light units
  • GPC3 binders showed in vitro anti-tumor activity against GPC3-expressing tumor cells.
  • Epitope 1, 3 and 4 on GPC3 were capable of activating CAR T-cells leading to cytotoxicity against tumor cells.
  • All GPC3 binders had improved or comparable cytotoxicity against Nalm-6 GPC3 + and Huh-7 tumor cells as compared to GC33 CAR T-cells.
  • Binders 1, 4, 8, 9 and 16 showed superior cytotoxicity against PLC/PRF/5 tumor cells as compared to GC33_28z1XX CAR T-cells.
  • Body weight was measured 3 times a week to assess the acute and severe toxicities associated with the treatment. All animals were sacrificed according to IACUC protocol standards. An experimental layout was illustrated in Figure 7.
  • CAR-T groups with binder 8 also referred to as “8_28z1XX (vH-vK)” and binder 16 (also referred to as “16_28z1XX (vH-vK)”) efficiently reduced tumor size over time and prolonged the overall survival of tumor bearing mice. All mice in CAR-T group with binder 8 achieved tumor free and sustained until termination of the study, while 3 of the 5 mice from binder 16 group achieved tumor free. See Figure 9.
  • mice in CAR-T group with binder 16 started to develop GvHD related symptoms including scaly and NAI-1540294631v3 240 inflamed skin at extremities, while mice in CAR-T group with binder 8 didn’t exhibit any similar symptoms. All these data suggest that binder 8 and binder 16 CAR-T cells could efficiently control tumor growth in vivo. Lower doses can be further assessed to explore the antitumor potency of the CAR-T and mitigate GvHD issues.
  • Body weight was measured 3 times a week to assess the acute and severe toxicities associated with the treatment. Blood samples were collected on day 7, day 14 post CAR-T injection and termination day of the study. The presence of human CAR-T cells was assessed by flow cytometry and IFN- ⁇ secretion was quantified by ELISA. All animals were sacrificed according to IACUC protocol standards. An experimental layout was illustrated in Figure 10. Further, peripheral blood from mice treated with CAR-T cells was collected on Day 51 and concentration of IFN- ⁇ in the plasma was quantified by ELISA.
  • CAR- T cells comprising binder 8 demonstrated potent anti-tumor efficacy and showed a minimal risk of toxicity in vivo.
  • the results of the restimulation assay indicate that the GPC3-targeted CAR T cells with binders 1, 2, 4, 8, 9, and 16 (corresponding to 1_28z1XX, 2_28z1XX, 4_28z1XX, 8_28z1XX, 9_28z1XX, and 16_28z1XX) can repeatedly kill tumor cells and continue to proliferate in response to antigen, respectively.
  • the GPC3-targeted CAR T cells exhibit higher cytotoxicity over four restimulation cycles compared to GC33 and Y035 CAR T cells.
  • the GPC3-targeted CAR T cells with binders 1, 8, 9 and 16 show higher CAR T-cell numbers over four restimulation cycles and continue to proliferate in response to Huh7 tumor cells.
  • all 6 GPC3-targeted CAR T cells showed equal or increased cytotoxicity and proliferation compared to the GPC3-targeted CAR T cells, while the comparator Y035 CAR T cells showed poor cytotoxicity and proliferation when co- cultured in the restimulation assays.
  • HCC hepatocellular carcinoma
  • the GPC3-targeted CAR T cells with binders 1, 9 and 16 were chosen to represent a range of affinities and epitope binding scFv binders. All CAR T-cells showed anti-tumor efficacy at both doses. See Figure 16.
  • Four mice treated with 2 ⁇ 10 5 16_28z1XX and 2 mice treated with GC33 CAR T cells had long term tumor free survival.
  • Mice treated with the higher dose of 5 ⁇ 10 5 9_28z1XX and 16_28z1XX CAR T cells had long term anti-tumor activity equal to or greater than GC33_28z1XX.
  • the CPTAC performed the first proteogenomic characterization of HBV- related hepatocellular carcinoma using paired tumor and adjacent liver tissues from 159 patients, revealing insights into multi-omics consistency, key signaling pathways, and liver-specific metabolic reprogramming.
  • proteomic (bulk) expression of GPC3 log2(fold change) of protein data was calculated by comparing tumor and normal liver samples.
  • Molecules were filtered to show only those found on the cell surface (The UniProt Consortium et al., 2023, Nucleic Acids Research 51 (D1): D523–D531; Thul et al., 2017, Science 356, eaal3321; Gene Ontology Consortium et al., 2023, Genetics 224, 1; Binder et. al., 2014, Database 2014) and over-expressed in tumor compared to liver. Log2FC of the filtered molecules were percentile ranked and their distribution plotted. Density plots were created to visualize the distribution of protein expression levels.
  • Transcriptomic (bulk) expression of GPC3 was analyzed using the data pulled from The Cancer Genome Atlas (TCGA) and the CPTAC.
  • the TCGA aimed to comprehensively analyze 363 HCC cases using whole-exome sequencing, DNA copy number analyses, and 196 HCC cases using DNA methylation, RNA, miRNA, and proteomic expression, identifying significantly mutated genes and molecular subtypes (Ally et al., 2017, Cell 169 (7): 1327 - 1341.e23).
  • the goal for using the HCC CPTAC data was the same as mentioned in the analysis of the proteomic (bulk) expression of GPC3.
  • GPC3 In the analysis on transcriptomic (bulk) expression of GPC3, log2(fold change) of transcript data was calculated by comparing tumor and normal liver samples. Transcripts were filtered to show only those coding for membrane-bound proteins and overexpressed in tumor compared to liver. Log2FC of the filtered transcripts were percentile ranked and their distribution plotted. Density plots were created to visualize the distribution of transcript expression levels. [00839] As shown in Figure 19A, GPC3 is identified as the most over-expressed membrane- bound protein when comparing tumor versus normal liver samples in both the CNHPP and NAI-1540294631v3 244 CPTAC datasets.
  • GPC3 is among the most highly over-expressed transcripts when comparing tumor versus normal liver samples in both the TCGA and CPTAC datasets.

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Abstract

The presently disclosed subject matter provides binding molecules for glypican-3 (GPC3), including antibodies or antigen-binding fragments thereof that bind to GPC3 and antigen recognizing receptors (e.g., CARs) that target GPC3. Also provided are cells and compositions comprising such binding molecules, and methods for treating diseases or disorders associated with GPC3, e.g., hepatocellular carcinoma.

Description

Attorney Docket No.14259-055-228 GPC3-TARGETED MOLECULES AND USES THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No.63/510,323, filed June 26, 2023, the disclosure of which is incorporated by reference herein it its entirety. SEQUENCE LISTING This application contains a computer readable Sequence Listing which has been submitted in XML file format with this application, the entire content of which is incorporated by reference herein in its entirety. The Sequence Listing XML file submitted with this application is entitled “14259-055-228_SEQLISTING.xml”, was created on June 21, 2024, and is 495,078 bytes in size. 1. TECHNICAL FIELD [0001] The presently disclosed subject matter provides molecules that target glypican-3 (GPC3), cells comprising the molecules, and methods for treating GPC3-related diseases using such molecules and cells. 2. BACKGROUND [0002] Cell-based immunotherapy is a therapy with curative potential for the treatment of cancer. T-cells and other immune cells may be modified to target tumor antigens through the introduction of genetic material coding for artificial or synthetic receptors for antigen, termed Chimeric Antigen Receptors (CARs), specific to selected antigens. Targeted T-cell therapy using CARs has shown recent clinical success in treating hematologic malignancies. [0003] GPC3 is widely expressed in the placenta and fetal tissues, while its expression is significantly reduced in adult organs. In addition, significantly high expression levels of GPC3 have been found in hepatocellular carcinoma (HCC) cells compared to normal liver and non-neoplastic liver lesions. Thus, GPC3 is a promising tumor marker and a potential molecular target for therapeutic intervention in HCC. Thus, there are needs for GPC3-targeted molecules, e.g., CARs or antibodies, for treating tumors, e.g., HCC. 3. SUMMARY OF THE INVENTION [0004] The presently disclosed subject matter provides GPC3-targeted molecules (e.g., antibodies, antigen-binding fragments, chimeric antigen receptor (CAR)s), cells comprising the GPC3-targeted molecules, and uses of the cells and antibodies, e.g., for treating a GPC3-related disease or disorder. NAI-1540294631v3 1 [0005] The presently disclosed subject matter provides GPC3-targeted CARs. In certain embodiments, the GPC3-targeted CAR comprises an extracellular domain that binds to GPC3, a transmembrane domain, and an intracellular domain, wherein the extracellular domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: (a) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 30; (b) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 47, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 48; (c) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 62, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 63; (d) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 75, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 76; (e) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 89, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 90; (f) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 101, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 102; (g) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 119, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 120; (h) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 137, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 138; (i) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 155, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 156; NAI-1540294631v3 2 (j) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 170, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 171; (k) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 183, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 184; (l) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 201, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 202; (m) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 215, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 216; (n) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 233, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 234; (o) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 246, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 247; or (p) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 262, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 263. [0006] In certain embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system, the Chothia numbering system, the AbM numbering system, the Contact numbering system, or the IMGT® Information System. In certain embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system. [0007] In certain embodiments, the VH and VL are characterized below: (a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative NAI-1540294631v3 3 modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof; (b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (c) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (d) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (e) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative NAI-1540294631v3 4 modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (f) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (g) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof; (h) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof; (i) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative NAI-1540294631v3 5 modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof; (j) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof; (k) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (l) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof; (m) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a NAI-1540294631v3 6 conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof; (n) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof; (o) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof; or (p) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof. [0008] In certain embodiments, the VH and VL are characterized below: (a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28; NAI-1540294631v3 7 (b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (c) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (d) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (e) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (f) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (g) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the NAI-1540294631v3 8 amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118; (h) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136; (i) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154; (j) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169; (k) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (l) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200; NAI-1540294631v3 9 (m) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169; (n) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232; (o) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118; or (p) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. [0009] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136. [0010] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL NAI-1540294631v3 10 comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. [0011] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262. [0012] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262. [0013] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263. [0014] In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263. [0015] In certain embodiments, the VH and VL are characterized below: (a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; (b) the VH comprises the amino acid sequence set forth in SEQ ID NO: 47; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 48; (c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 62; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 63; NAI-1540294631v3 11 (d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; (e) the VH comprises the amino acid sequence set forth in SEQ ID NO: 89; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 90; (f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 101; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 102; (g) the VH comprises the amino acid sequence set forth in SEQ ID NO: 119; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 120; (h) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; (i) the VH comprises the amino acid sequence set forth in SEQ ID NO: 155; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 156; (j) the VH comprises the amino acid sequence set forth in SEQ ID NO: 170; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 171; (k) the VH comprises the amino acid sequence set forth in SEQ ID NO: 183; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 184; (l) the VH comprises the amino acid sequence set forth in SEQ ID NO: 201; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 202; (m) the VH comprises the amino acid sequence set forth in SEQ ID NO: 215; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 216; (n) the VH comprises the amino acid sequence set forth in SEQ ID NO: 233; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 234; (o) the VH comprises the amino acid sequence set forth in SEQ ID NO: 246; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 247; or (p) the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263. [0016] In certain embodiments, the VH and VL are characterized below: (a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; (b) the VH comprises the amino acid sequence set forth in SEQ ID NO: 47; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 48; (c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; NAI-1540294631v3 12 (d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; (e) the VH comprises the amino acid sequence set forth in SEQ ID NO: 155; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 156; or (f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263. [0017] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138. [0018] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263. [0019] In certain embodiments, the VH and the VL are connected via a linker. In certain embodiments, the linker consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4. In certain embodiments, the linker consists of the amino acid sequence set forth in SEQ ID NO: 1. [0020] In certain embodiments, the VH and the VL are positioned from the N- to the C-terminus: VL-VH. In certain embodiments, the VH and the VL are positioned from the N- to the C-terminus: VH-VL. [0021] In certain embodiments, the extracellular domain comprises a variable fragment (Fv), a single-chain variable fragment (scFv), a Fab, or a F(ab)2. In certain embodiments, the extracellular domain comprises an scFv. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO: 264, or SEQ ID NO: 265. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 264, or SEQ ID NO: 265. [0022] In certain embodiments, the transmembrane domain comprises a CD8 polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3 NAI-1540294631v3 13 polypeptide, a 2B4 polypeptide, or a BTLA polypeptide. In certain embodiments, the transmembrane domain comprises a CD28 polypeptide. In certain embodiments, the CD28 polypeptide comprises amino acids 154 to 179 of the amino acid sequence set forth in SEQ ID NO: 308. [0023] In certain embodiments, the intracellular domain comprises a CD3ζ polypeptide. In certain embodiments, the CD3ζ polypeptide is a modified CD3ζ polypeptide. In certain embodiments, the modified CD3ζ polypeptide comprises a native ITAM1, an ITAM2 variant consisting of two loss-of-function mutations, and an ITAM3 consisting of two loss-of-function mutations. In certain embodiments, the native ITAM1 consists of the amino acid sequence set forth in SEQ ID NO: 313. In certain embodiments, the ITAM2 variant consists of the amino acid sequence set forth in SEQ ID NO: 319. In certain embodiments, the ITAM3 variant consists of the amino acid sequence set forth in SEQ ID NO: 323. In certain embodiments, the modified CD3ζ polypeptide comprises or consists of the amino acid sequence set forth in SEQ ID NO: 325. In certain embodiments, the intracellular domain further comprises at least one co-stimulatory signaling region. In certain embodiments, the at least one co-stimulatory signaling region comprises at least one intracellular signaling domain of a co-stimulatory molecule. In certain embodiments, the co- stimulatory molecule is selected from the group consisting of CD28, 4-1BB, OX40, ICOS, DAP-10, CD27, CD28, CD30, CD40, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, CD8, CD4, B2C, CD80, CD86, DAP10, DAP12, MyD88, BTNL3, and NKG2D, and combinations thereof. In certain embodiments, the co-stimulatory molecule is CD28. In certain embodiments, the at least one co-stimulatory signaling region comprises amino acids 180 to 220 of the amino acid sequence set forth in SEQ ID NO: 308. [0024] In certain embodiments, the CAR further comprises a signal peptide. In certain embodiments, the signal peptide comprises the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the CAR further comprises a self-cleaving P2A peptide. In certain embodiments, the self-cleaving P2A peptide comprises the amino acid sequence set forth in SEQ ID NO: 329. [0025] In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 333, SEQ ID NO: 335, SEQ ID NO: 337, SEQ ID NO: 339, SEQ ID NO: 341, SEQ ID NO: 343, SEQ ID NO: 345, SEQ ID NO: 347, SEQ ID NO: 349, SEQ ID NO: 351, SEQ ID NO: 353, or SEQ ID NO: 355. NAI-1540294631v3 14 [0026] In certain embodiments, the CAR is expressed from a vector. In certain embodiments, the vector is a viral vector. In certain embodiments, the viral vector is a retroviral vector. [0027] The presently disclosed subject matter further provides nucleic acids encoding the GPC3- targeted CARs disclosed herein. In certain embodiments, the nucleic acid further comprises a promoter that is operably linked to the CAR. In certain embodiments, the promoter is endogenous or exogenous. In certain embodiments, the exogenous promoter is selected from the group consisting of an elongation factor (EF)-1 promoter, a cytomegalovirus immediate-early promoter (CMV) promoter, a simian virus 40 early promoter (SV40) promoter, a phosphoglycerate kinase (PGK) promoter, a metallothionein promoter, and Ubiquitin C promoter. In certain embodiments, the endogenous promoter is selected from the group consisting of a TCR alpha promoter, a TCR beta promoter, and a beta 2-microglobulin promoter. In certain embodiments, the promoter is an inducible promoter. In certain embodiments, the inducible promoter is selected from the group consisting of a NFAT transcriptional response element (TRE) promoter, a CD69 promoter, a CD25 promoter, an IL-2 promoter, a 4-1BB promoter, a PD1 promoter, and a LAG3 promoter. In certain embodiments, the nucleic acid comprises the nucleotide sequence set forth in SEQ ID NO: 334, SEQ ID NO: 336, SEQ ID NO: 338, SEQ ID NO: 340, SEQ ID NO: 342, SEQ ID NO: 344, SEQ ID NO: 346, SEQ ID NO: 348, SEQ ID NO: 350, SEQ ID NO: 352, SEQ ID NO: 354, or SEQ ID NO: 356. [0028] The presently disclosed subject matter provides vectors comprising the nucleic acids disclosed herein. In certain embodiments, the vector is a viral vector. In certain embodiments, the viral vector is a retroviral vector. [0029] The presently disclosed subject matter further provides cells comprising the CARs, nucleic acids, or vectors disclosed herein. [0030] In certain embodiments, the cell is an immunoresponsive cell or an immune effector cell. In certain embodiments, the cell is a cell of the lymphoid lineage or a cell of the myeloid lineage. In certain embodiments, the cell is selected from the group consisting of a T cell, a B cell, a Natural Killer (NK) cell, macrophage, an innate lymphoid cell (ILC), a cytokine induced killer (CIK) cell, a lymphokine activated killer (LAK) cell, a stem cell from which a lymphoid cell may be differentiated, a stem cell from which a myeloid cell may be differentiated, or a combination thereof. In certain embodiments, the cell is a T cell. In certain embodiments, the T cell is selected from the group consisting of helper T cells, cytotoxic T cells, memory T cells, regulatory T cells, tumor- infiltrating lymphocyte (TIL), Natural Killer T cells, mucosal associated invariant T cells, αβ T cells, and γδ T cells. In certain embodiments, the cell is a NK cell. In certain embodiments, the NK cell is NAI-1540294631v3 15 derived from a stem cell. In certain embodiments, the stem cell is a pluripotent stem cell. In certain embodiments, the pluripotent stem cell is an embryoid stem cell or an induced pluripotent stem cell. [0031] Furthermore, the presently disclosed subject matter provides anti-GPC3 antibodies or antigen-binding fragments thereof. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein: (a) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 30; (b) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 47, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 48; (c) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 62, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 63; (d) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 75, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 76; (e) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 89, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 90; (f) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 101, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 102; (g) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 119, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 120; (h) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 137, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 138; (i) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 155, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 156; NAI-1540294631v3 16 (j) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 170, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 171; (k) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 183, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 184; (l) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 201, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 202; (m) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 215, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 216; (n) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 233, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 234; (o) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 246, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 247; or (p) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 262, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 263. [0032] In certain embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system, the Chothia numbering system, the AbM numbering system, the Contact numbering system, or the IMGT® Information System. In certain embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system. [0033] In certain embodiments, the VH and VL are characterized as below: (a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative NAI-1540294631v3 17 modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof; (b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (c) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (d) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (e) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative NAI-1540294631v3 18 modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (f) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (g) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof; (h) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof; (i) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative NAI-1540294631v3 19 modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof; (j) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof; (k) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; (l) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof; (m) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a NAI-1540294631v3 20 conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof; (n) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof; (o) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof; or (p) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof. [0034] In certain embodiments, the VH and VL are characterized as below: (a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28; NAI-1540294631v3 21 (b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (c) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (d) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (e) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (f) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (g) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the NAI-1540294631v3 22 amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118; (h) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136; (i) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154; (j) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169; (k) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; (l) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200; NAI-1540294631v3 23 (m) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169; (n) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232; (o) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118; or (p) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. [0035] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136. [0036] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL NAI-1540294631v3 24 comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. [0037] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262. [0038] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263. [0039] In certain embodiments, the VH and VL are characterized as below: (a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; (b) the VH comprises the amino acid sequence set forth in SEQ ID NO: 47; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 48; (c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 62; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 63; NAI-1540294631v3 25 (d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; (e) the VH comprises the amino acid sequence set forth in SEQ ID NO: 89; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 90; (f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 101; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 102; (g) the VH comprises the amino acid sequence set forth in SEQ ID NO: 119; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 120; (h) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; (i) the VH comprises the amino acid sequence set forth in SEQ ID NO: 155; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 156; (j) the VH comprises the amino acid sequence set forth in SEQ ID NO: 170; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 171; (k) the VH comprises the amino acid sequence set forth in SEQ ID NO: 183; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 184; (l) the VH comprises the amino acid sequence set forth in SEQ ID NO: 201; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 202; (m) the VH comprises the amino acid sequence set forth in SEQ ID NO: 215; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 216; (n) the VH comprises the amino acid sequence set forth in SEQ ID NO: 233; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 234; (o) the VH comprises the amino acid sequence set forth in SEQ ID NO: 246; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 247; or (p) the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263. [0040] In certain embodiments, the VH and VL are characterized as below: (a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; (b) the VH comprises the amino acid sequence set forth in SEQ ID NO: 47; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 48; (c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; NAI-1540294631v3 26 (d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; (e) the VH comprises the amino acid sequence set forth in SEQ ID NO: 155; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 156; or (f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263. [0041] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138. [0042] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263. [0043] In certain embodiments, the antigen-binding fragment thereof is a variable fragment (Fv), a single-chain variable fragment (scFv), a Fab, or a F(ab)2. In certain embodiments, the antigen- binding fragment thereof is an scFv. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO: 264, or SEQ ID NO: 265. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 264, or SEQ ID NO: 265. [0044] In certain embodiments, the VH is connected to the VL via a linker. In certain embodiments, the linker consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4. In certain embodiments, the linker consists of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the VH and the VL are positioned from the N- to the C-terminus: VL-VH. In certain embodiments, the VH and the VL are positioned from the N- to the C-terminus: VH-VL. [0045] In certain embodiments, the antibody comprises a human variable region framework region. In certain embodiments, the antibody or antigen-binding fragment thereof comprises a Fc region. In certain embodiments, the Fc region comprises the amino acid sequence set forth in SEQ ID NO: 357. NAI-1540294631v3 27 [0046] In certain embodiments, the antibody or antigen-binding fragment thereof is a fully human or an antigen-binding fragment thereof, a chimeric antibody or an antigen-binding fragment thereof, or a humanized antibody or an antigen-binding fragment thereof. [0047] The presently disclosed subject matter further provides antibodies or antigen-binding fragments thereof that compete with any one of the anti-GPC3 antibodies or antigen-binding fragments thereof disclosed herein for binding to GPC3. And, the presently disclosed subject matter provides antibodies or antigen-binding fragments thereof that bind to essentially the same epitope region on GPC3 as any one of the anti-GPC3 antibodies or antigen-binding fragments thereof disclosed herein. [0048] The presently disclosed subject matter further provides conjugates comprising the antibodies or antigen-binding fragments thereof disclosed herein. In certain embodiments, the antibody or antigen-binding fragment thereof is linked to a therapeutic agent, a detectable agent, or a diagnostic agent. In certain embodiments, the therapeutic agent is a chemotherapeutic agent, a cytotoxin, or a drug. In certain embodiments, the conjugate is an immunoconjugate. [0049] The presently disclosed subject matter provides multispecific molecules comprising any one of the antibodies or antigen-binding fragments thereof disclosed herein that is linked to a second functional moiety. In certain embodiments, the second functional moiety has a different binding specificity than the antibody or antigen binding fragment thereof. [0050] Furthermore, the presently disclosed subject matter provides nucleic acids that encode the antibodies or antigen-binding fragments thereof. In certain embodiments, the nucleic acid comprises a first polynucleotide that encodes a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262; and/or a second polynucleotide that encodes a VL comprising the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263. [0051] In certain embodiments, the first polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO: 33, SEQ ID NO: 51, SEQ ID NO: 66, SEQ ID NO: 79, SEQ ID NO: 93, SEQ ID NO: 105, SEQ ID NO: 123, SEQ ID NO: 141, SEQ ID NO: 159, SEQ ID NO: 174, SEQ ID NO: 187, SEQ ID NO: 205, SEQ ID NO: 219, SEQ ID NO: 237, SEQ ID NO: 250, or SEQ ID NO: 266. In certain embodiments, the second polynucleotide comprises the nucleotide sequence set forth in NAI-1540294631v3 28 SEQ ID NO: 35, SEQ ID NO: 53, SEQ ID NO: 68, SEQ ID NO: 81, SEQ ID NO: 95, SEQ ID NO: 107, SEQ ID NO: 125, SEQ ID NO: 143, SEQ ID NO: 161, SEQ ID NO: 176, SEQ ID NO: 189, SEQ ID NO: 207, SEQ ID NO: 221, SEQ ID NO: 239, SEQ ID NO: 252, or SEQ ID NO: 268. [0052] The presently disclosed subject matter provides vectors comprising the nucleic acids disclosed herein. In certain embodiments, the vector is an expression vector. [0053] The presently disclosed subject matter further provides host cells comprising the vectors disclosed herein. [0054] The presently disclosed subject matter provides methods of producing a presently disclosed anti-GPC3 antibody or an antigen-binding fragment thereof. In certain embodiments, the method comprises culturing a presently disclosed host cell under conditions to induce expression of the antibody or antigen-binding fragment thereof from the host cell. [0055] The presently disclosed subject matter provides methods for detecting GPC3 in a whole cell or tissue. In certain embodiments, the method comprises: (a) contacting a cell or tissue with a presently disclosed anti-GPC3 antibody or an antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises a detectable label; and (b) determining the amount of the labeled antibody or antigen-binding fragment thereof bound to the cell or tissue by measuring the amount of detectable label associated with the cell or tissue, wherein the amount of bound antibody or antigen-binding fragment thereof indicates the amount of GPC3 in the cell or tissue. [0056] The presently disclosed composition comprises the cells, the antibodies or antigen- binding fragments, the conjugates, or the multispecific molecules disclosed herein. In certain embodiments, the composition is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier. In certain embodiments, the composition comprises between about 1 × 105 and about 5 × 108 cells. [0057] The presently disclosed cells, antibodies or antigen-binding fragments thereof, conjugates, multispecific molecules, or compositions can be used in a therapy. [0058] The presently disclosed cells, antibodies or antigen-binding fragments thereof, conjugates, multispecific molecules, or compositions can be used in a method for treating a disease or disorder associated with GPC3 in a subject. [0059] The presently disclosed subject matter provides methods of treating a disease or disorder associated with GPC3 in a subject. In certain embodiments, the method comprises: administering to the subject cells comprising a presently disclosed GPC3-targeted CAR, the anti-GPC3 antibody or NAI-1540294631v3 29 antigen-binding fragment thereof disclosed herein, the conjugate disclosed herein, the multispecific molecule disclosed herein, or the composition disclosed herein. [0060] The presently disclosed subject matter provides uses of the presently disclosed cells, antibodies or antigen-binding fragments thereof, conjugates, multispecific molecules, or compositions in the manufacture of a medicament for treating a disease or disorder associated with GPC3. [0061] In certain embodiments, the disease or disorder is a tumor. In certain embodiments, the disease or disorder is cancer. In certain embodiments, the disease or disorder is selected from the group consisting of hepatocellular carcinoma, hepatoblastoma, lung squamous cell carcinoma, ovarian yolk sac tumor, melanoma, and urothelial carcinoma. In certain embodiments, the disease or disorder is hepatocellular carcinoma. In certain embodiments, the subject is a human subject. [0062] The presently disclosed subject matter provides methods for producing a cell comprising a presently disclosed GPC3-targeted CAR. In certain embodiments, the method comprises: introducing into a cell a nucleic acid that encodes the presently disclosed GPC3-targeted CAR. 4. BRIEF DESCRIPTION OF THE DRAWINGS [0063] Figures 1A to 1E depict the binding activity of scFv-Fc fusion proteins of 16 binders to cells expressing human GPC3 using fluorescence-activated cell sorting (FACS) analysis. Each of the purified scFv-Fc proteins starting at a concentration of 45 μg/mL was serially diluted 12 times with a dilution factor of three, and incubated with CHO-K1/human GPC3 clone 9(+) cells expressing human GPC3. An anti-hIgG GPC3 antibody was used as a positive control, and a human IgG was used as an isotype control, both of which were diluted in the same manner as the tested proteins. A PBS solution was used as a negative control. Each of the purified scFv-Fc fusion proteins was also tested for binding to a negative control cell line, CHO-K1(-). Plates were read by flow cytometry reader. Binding curves were plotted using the geometric-mean fluorescence intensity of the fluorescence signal on the cells, and the EC50 values were calculated. [0064] Figures 2A and 2B depict the generation of the presently disclosed GPC3-targeted scFv CAR T Cells. Figure 2A depicts structures of CARs in accordance with certain embodiments of the presently disclosed subject matter. Figure 2B depicts the flow cytometric analysis showing the expression levels of LNGFR for the indicated constructs in the VH-VK and VK-VH orientation. [0065] Figure 3 depicts the GPC3 expression levels of Nalm-6 GPC3+, Huh7, and PLC/PRF/5 tumor cells. [0066] Figures 4A and 4B depict the in vitro cytotoxicity of the presently disclosed GPC3- targeted CAR T cells against Nalm-6 GPC3+ and Nalm-6 WT tumor cells. Figure 4A shows the NAI-1540294631v3 30 results of 16 hour Cytotoxic T lymphocyte (CTL) mediated cytotoxicity using Nalm-6 GPC3+ tumor cells. Figure 4B shows the results of 16 hour CTL using Nalm-6 wildtype (WT) tumor cells. [0067] Figure 5 depicts the in vitro cytotoxicity of the presently disclosed GPC3-targeted CAR T cells against Huh-7 tumor cells. Figure 5 shows the results of 16 hour CTL using Huh-7 tumor cells. [0068] Figure 6 depicts the in vitro cytotoxicity of the presently disclosed GPC3-targeted CAR T cells against PLC/PRF/5 tumor cells. Figure 6 shows the results of 16 hour CTL using PLC/PRF/5 tumor cells. [0069] Figure 7 depicts an experimental layout of an assessment of the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells. [0070] Figure 8 depicts the body weight changes of mice who received the presently disclosed GPC3-targeted CAR T cells. [0071] Figure 9 depicts the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells. [0072] Figure 10 depicts an experimental layout of an assessment of the in vivo safety and effective dose of the presently disclosed GPC3-targeted CAR with binders 8 and 16, respectively. [0073] Figure 11 depicts the survival rates of mice treated with the presently disclosed GPC3- targeted CAR with binders 8 and 16, respectively, at difference doses. [0074] Figure 12 depicts body weight changes of mice treated with the presently disclosed GPC3-targeted CAR with binders 8 and 16, respectively, at difference doses. [0075] Figure 13 depicts the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR with binders 8 and 16, respectively, at difference doses. [0076] Figure 14 depicts the safety of the presently disclosed GPC3-targeted CAR with binders 8 and 16, respectively, at difference doses. [0077] Figures 15A to 15C depict the restimulation assay with Huh7 tumor cells. Figure 15A depicts a scheme showing the principle of CAR T restimulation assay. Figure 15B shows the results of the Huh7 tumor cell lysis of the VH-VK GPC3 CAR T cells. Figure 15C shows the results of the proliferation of the CAR T cells. [0078] Figure 16 depicts an assessment of the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells to orthotopically implanted Huh7 tumor cells in liver. [0079] Figure 17 depicts an assessment of the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells to subcutaneously implanted Huh7 tumor cells. [0080] Figure 18 depicts an assessment of the in vivo antitumor efficacy of the presently disclosed GPC3-targeted CAR T cells to Nalm-6 GPC3+ tumor cells injected via tail vein. NAI-1540294631v3 31 [0081] Figures 19A and 19B depict an analysis of GPC3 expression level in tumor liver samples versus normal liver samples. Figure 19A shows the density plot of upregulated membrane-bound proteins from mass spectrometry data obtained from the Chinese Human Proteome Project (CNHPP) (Jiang et al., 2019, Nature 567: 257–261) and the Clinical Proteomic Tumor Analysis Consortium (CPTAC) (Gao et al., 2019, Cell 179 (2): 561 - 577.e22), respectively. Proteins below the 98th percentile ranking of expression are colored in light color, and those in and above the 98th percentile ranking are colored in dark color. Figure 19B shows the density plot of upregulated membrane- bound transcripts from bulk RNAseq data obtained from The Cancer Genome Atlas (TCGA) (Ally et al., 2017, Cell 169 (7): 1327 - 1341.e23) and the CPTAC. Transcripts below the 98th percentile ranking of expression are colored in light color, and those in and above the 98th percentile ranking are colored in dark color. 5. DETAILED DESCRIPTION [0082] The presently disclosed subject matter provides molecules that target GPC3 (referred to as “GPC3-targeted molecules”). In certain embodiments, the molecule is an antibody or an antigen- binding fragment thereof. In certain embodiments, the molecule is an antigen recognizing receptor. In certain embodiments, the antigen recognizing receptor is a chimeric antigen receptor (CAR). The presently disclosed subject matter further discloses cells comprising such GPC3-targeted molecules. The cells can be immunoresponsive cells and/or immune effector cells, e.g., genetically modified immunoresponsive cells (e.g., T cells or NK cells), or pluripotent stem cell that can differentiate into immunoresponsive cells and/or immune effector cells. [0083] Non-limiting embodiments of the present disclosure are described by the present specification and Examples. [0084] For purposes of clarity of disclosure and not by way of limitation, the detailed description is divided into the following subsections: 5.1. Definitions; 5.2. GPC3; 5.3. Chimeric Antigen Receptors (CARs); 5.4. Cells; 5.5. Anti-GPC3 Antibodies; 5.6. Nucleic Acids and Vectors; 5.7. Methods of Detection; 5.8. Formulations and Administration; and 5.9. Methods of Treatment NAI-1540294631v3 32 5.1. Definitions [0085] Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which the presently disclosed subject matter belongs. [0086] As used herein, the term “about” or “approximately” refers to within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, “about” can mean within 3 or more than 3 standard deviations, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, and more preferably still up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold, of a value. [0087] As used herein, the term “immunoresponsive cell” refers to a cell that functions in an immune response or a progenitor, or progeny thereof. In certain embodiments, the immunoresponsive cell is a cell of lymphoid lineage. Non-limiting examples of cells of lymphoid lineage include T cells, Natural Killer (NK) cells, B cells, and stem cells from which lymphoid cells may be differentiated. In certain embodiments, the immunoresponsive cell is a cell of myeloid lineage. [0088] As used herein, the term “activates an immunoresponsive cell” refers to induction of signal transduction or changes in protein expression in the cell resulting in initiation of an immune response. For example, when CD3 chains cluster in response to ligand binding and immunoreceptor tyrosine-based inhibition motifs (ITAMs) a signal transduction cascade is produced. In certain embodiments, when a CAR binds to an antigen, a formation of an immunological synapse occurs that includes clustering of many molecules near the bound receptor (e.g., CD4 or CD8, CD3γ/δ/є/ξ, etc.). This clustering of membrane bound signaling molecules allows for ITAM motifs contained within the CD3 chains to become phosphorylated. This phosphorylation in turn initiates a T cell activation pathway ultimately activating transcription factors, such as NF-κB and AP-1. These transcription factors induce global gene expression of the T cell to increase IL-2 production for proliferation and expression of master regulator T cell proteins in order to initiate a T cell mediated immune response. [0089] As used herein, the term “stimulates an immunoresponsive cell” refers to a signal that results in a robust and sustained immune response. In certain embodiments, this occurs after NAI-1540294631v3 33 immunoresponsive cell (e.g., T cell) activation or concomitantly mediated through receptors including, but not limited to, CD28, 4-1BB, OX40, CD40 and ICOS. Receiving multiple stimulatory signals can be important to mount a robust and long-term T-cell mediated immune response. T cells can quickly become inhibited and unresponsive to antigen. While the effects of these co-stimulatory signals may vary, they generally result in increased gene expression in order to generate long lived, proliferative, and anti-apoptotic T cells that robustly respond to antigen for complete and sustained eradication. [0090] The term “antibody” covers, for example polyclonal antibodies, monoclonal antibodies (including agonist, antagonist, neutralizing antibodies, full length monoclonal antibodies), antibody compositions with polyepitopic or monoepitopic specificity, recombinantly produced antibodies, monospecific antibodies, multispecific antibodies (including bispecific antibodies), synthetic antibodies, chimeric antibodies, humanized antibodies, or human versions of antibodies having full length heavy and/or light chains. The present disclosure also includes antibody fragments (and/or polypeptides that comprise antibody fragments) that retain α5 integrin binding characteristics. Non- limiting examples of antibody fragments include antigen-binding regions and/or effector regions of the antibody, e.g., Fab, Fab’, F(ab’)2, Fv, scFv, (scFv)2, single chain antibody molecule, dual variable region antibody, single variable region antibody, linear antibody, V region, a multispecific antibody formed from antibody fragments, F(ab)2, Fd, Fc, diabody, di-diabody, disulfide-linked Fvs (dsFv), single-domain antibody (e.g., VHH, nanobody) or other fragments (e.g., fragments consisting of the variable regions of the heavy and light chains that are non-covalently coupled). In general terms, a variable region domain may be any suitable arrangement of immunoglobulin heavy (VH) and/or light (VL) variable regions. For example, the presently disclosed subject matter also includes tetrameric antibodies comprising two heavy chain and two light chain molecules, an antibody light chain monomer, and an antibody heavy chain monomer. Thus, for example, the variable region domain may be dimeric and contain VH-VH, VH-VL, or VL-VL dimers that bind α5 integrin. If desired, the VH and VL chains may be covalently coupled either directly or through a linker to form a single chain Fv (scFv). For ease of reference, scFv proteins are referred to herein as included in the category “antibody fragments.” Another form of an antibody fragment is a peptide comprising one or more complementarity determining regions (CDRs) of an antibody. CDRs can be obtained by constructing polynucleotides that encode the CDR of interest. Such polynucleotides are prepared, for example, by using the polymerase chain reaction to synthesize the variable region using mRNA of antibody-producing cells as a template (see, for example, Larrick et al., Methods: A Companion to Methods in Enzymology, 2:106 (1991); Courtenay-Luck, “Genetic Manipulation of NAI-1540294631v3 34 Monoclonal Antibodies,” in Monoclonal Antibodies Production, Engineering and Clinical Application, Ritter et al. (eds.), page 166, Cambridge University Press (1995); and Ward et al., “Genetic Manipulation and Expression of Antibodies,” in Monoclonal Antibodies: Principles and Applications, Birch et al., (eds.), page 137, Wiley-Liss, Inc. (1995)). Antibody fragments may be incorporated into single domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, variable domains of new antigen receptors (v-NAR), and bis-single chain Fv regions (see, e.g., Hollinger and Hudson, Nature Biotechnology, 23(9):1126-1136, 2005). The antibodies or antigen-binding fragments thereof, in certain embodiments, comprise a light chain and/or a heavy chain constant region, such as one or more constant regions, including one or more IgG1, IgG2, IgG3 and/or IgG4 constant regions. In certain embodiments, antibodies can include epitope-binding fragments of any of the above. The antibodies described herein can be of any class (e.g., IgG, IgE, IgM, IgD, and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) of immunoglobulin molecule. Antibodies can be antagonistic antibodies or agonistic antibodies. [0091] As used herein, the term “monospecific antibody” refers to an antibody that has one or more binding sites each of which bind to the same epitope of the same antigen, e.g., GPC3 (e.g., human GPC3). [0092] As used herein, the term “multispecific molecule” refers to a molecule (e.g., an antibody) that is capable of binding to at least two distinct antigenic determinants, for example two binding sites each formed by a pair of an antibody heavy chain variable domain (VH) and an antibody light chain variable domain (VL) binding to different antigens or to different epitopes on the same antigen. Such a multispecific molecule may have a 1+1 format. Other multispecific molecule formats may be 2+1 or 1+2 formats (comprising two binding sites for a first antigen or epitope and one binding site for a second antigen or epitope) or 2+2 formats (comprising two binding sites for a first antigen or epitope and two binding sites for a second antigen or epitope). When a multispecific molecule comprises two antigen binding sites, each may bind to a different antigenic determinant. Such a multispecific molecule may bind to two different epitopes on the same antigen (e.g., epitopes on GPC3) or on different antigens (e.g., an epitope on GPC3 and an epitope on an antigen different from GPC3). [0093] The term “epitope” refers to the region of an antigen to which an antibody binds preferentially and specifically. A monoclonal antibody binds preferentially to a single specific epitope of a molecule that can be molecularly defined. In the present invention, multiple epitopes can be recognized by a multispecific antibody. NAI-1540294631v3 35 [0094] As used herein, the term “antibody fragment” refers to any derivative of an antibody which is less than full-length. In exemplary embodiments, the antibody fragment retains at least a significant portion of the full-length antibody’s specific binding ability. Examples of antibody fragments include, but are not limited to, Fab, Fab′, F(ab′)2, scFv, Fv, dsFv diabody, Fc, and Fd fragments. The antibody fragment may be produced by any means. For instance, the antibody fragment may be enzymatically or chemically produced by fragmentation of an intact antibody, it may be recombinantly produced from a gene encoding the partial antibody sequence, or it may be wholly or partially synthetically produced. The antibody fragment may optionally be a single chain antibody fragment. Alternatively, the fragment may comprise multiple chains which are linked together, for instance, by disulfide linkages. The fragment may also optionally be a multimolecular complex. A functional antibody fragment will typically comprise at least about 50 amino acids and more typically will comprise at least about 200 amino acids. [0095] As used herein, the term “Fab fragment” refers to a fragment of an antibody comprising an antigen-binding site generated by cleavage of the antibody with the enzyme papain, which cuts at the hinge region N-terminally to the inter-H-chain disulfide bond and generates two Fab fragments from one antibody molecule. [0096] As used herein, the term “F(ab′)2 fragment” refers to a fragment of an antibody containing two antigen-binding sites, generated by cleavage of the antibody molecule with the enzyme pepsin which cuts at the hinge region C-terminally to the inter-H-chain disulfide bond. [0097] As used herein, the term “Fc fragment” refers to the fragment of an antibody comprising the constant domain of its heavy chain. [0098] As used herein, the term “Fv fragment” refers to the fragment of an antibody comprising the variable domains of its heavy chain and light chain. [0099] The term “single chain variable fragment” or “scFv” refers to an Fv fragment in which the heavy chain domain and the light chain domain are linked. One or more scFv fragments may be linked to other antibody fragments (such as the constant domain of a heavy chain or a light chain) to form antibody constructs having one or more antigen recognition sites. The heavy chain domain and light chain domain are either joined directly or joined by a linker, which connects the N-terminus of the heavy chain domain with the C-terminus of the light chain domain, or the C-terminus of the heavy chain domain with the N-terminus of the light chain domain. As used herein, the term “linker” is art-recognized and refers to a molecule or group of molecules connecting two NAI-1540294631v3 36 compounds, such as two polypeptides. The linker may be comprised of a single linking molecule or may comprise a linking molecule and a spacer molecule, intended to separate the linking molecule and a compound by a specific distance. The linker is usually rich in glycine for flexibility, as well as serine or threonine for solubility. In certain embodiments, the linker is a G4S linker. [00100] In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, which is provided below: GGGGSGGGGSGGGGS [SEQ ID NO: 1] [00101] In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 2, which is provided below: GGGGSGGGGSGGGSGGGGS [SEQ ID NO: 2] [00102] In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 3, which is provided below: GGGGSGGGGSGGGGSGGGSGGGGS [SEQ ID NO: 3] [00103] In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 4, which is provided below: GGGGSGGGGSGGGGSGGGGSGGGSGGGGS [SEQ ID NO: 4] [00104] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 1 is set forth in SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, or SEQ ID NO: 21, which is provided below. GGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCT [SEQ ID NO: 5] GGCGGCGGAGGCAGCGGCGGCGGCGGCTCTGGCGGCGGCGGAAGC [SEQ ID NO: 6] GGCGGAGGAGGCAGCGGCGGCGGAGGCTCTGGCGGCGGCGGAAGT [SEQ ID NO: 7] GGCGGCGGAGGCAGCGGCGGCGGCGGCTCCGGCGGAGGCGGCTCT [SEQ ID NO: 8] GGCGGCGGAGGCTCTGGAGGCGGAGGCAGCGGCGGCGGCGGCTCC [SEQ ID NO: 9] GGCGGCGGCGGCAGCGGCGGCGGAGGTTCTGGAGGAGGCGGCAGC [SEQ ID NO: 10] GGCGGCGGCGGCAGCGGCGGAGGCGGCTCTGGCGGAGGCGGCAGC [SEQ ID NO: 11] GGCGGAGGCGGATCTGGCGGCGGCGGTTCAGGCGGAGGCGGCTCT [SEQ ID NO: 12] GGCGGAGGCGGATCTGGCGGAGGCGGCTCTGGAGGCGGCGGCTCA [SEQ ID NO: 13] GGAGGCGGCGGATCCGGCGGTGGCGGATCTGGCGGAGGCGGCAGC [SEQ ID NO: 14] GGCGGAGGCGGCTCTGGAGGAGGAGGGTCCGGAGGCGGCGGCAGC [SEQ ID NO: 15] GGCGGCGGCGGCAGCGGCGGAGGCGGCAGCGGCGGCGGAGGCAGC [SEQ ID NO: 16] NAI-1540294631v3 37 GGCGGAGGCGGCAGCGGCGGCGGCGGAAGCGGCGGCGGAGGCAGC [SEQ ID NO: 17] GGCGGCGGAGGCTCTGGCGGTGGCGGAAGCGGCGGCGGCGGAAGC [SEQ ID NO: 18] GGAGGCGGCGGCAGCGGCGGCGGCGGCAGTGGCGGAGGCGGCAGC [SEQ ID NO: 19] GGCGGAGGCGGCAGCGGAGGAGGCGGCTCTGGCGGCGGCGGCTCA [SEQ ID NO: 20] GGCGGCGGCGGCAGCGGAGGCGGCGGCAGCGGAGGAGGTGGCAGC [SEQ ID NO: 21] [00105] As used herein, “complementarity determining regions” or “CDRs” are defined as the complementarity determining region amino acid sequences of an antibody which are the hypervariable regions of immunoglobulin heavy and light chains. CDRs can be identified according to a number of known numbering systems. In certain embodiments, the CDRs are identified according to the Kabat numbering system. The Kabat CDRs are based on sequence variability and are the most commonly used (see e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). The CDRs can be identified according to the Chothia numbering system. Chothia refers to the location of the structural loops (see e.g., Chothia and Lesk, J. Mol. Biol.196:901-917 (1987)). The CDRs can be identified according to the AbM numbering system. The AbM hypervariable regions represent a compromise between the Kabat CDRs and Chothia structural loops, and are used by Oxford Molecular’s AbM antibody modeling software (see e.g., Martin, in Antibody Engineering, Vol.2, Chapter 3, Springer Verlag). The CDRs can be identified according to the Contact numbering system (see, e.g., MacCallum RM et al., 1996, J Mol Biol 5: 732-745). The Contact CDRs are based on an analysis of the available complex crystal structures. The CDRs can be identified according to the ImMunoGeneTics (IMGT)® Information System. A universal numbering system has been developed and widely adopted, IMGT® Information System (Lefranc et al., Dev. Comp. Immunol. 27(1):55-77 (2003)). IMGT® is an integrated information system specializing in immunoglobulins (IG), T cell receptors (TR) and major histocompatibility complex (MHC) of human and other vertebrates. [00106] The term “amino acid sequence” refers to a list of abbreviations, letters, characters or words representing amino acid residues. The amino acid abbreviations used herein are conventional one letter codes for the amino acids and are expressed as follows: A, alanine; B, asparagine or aspartic acid; C, cysteine; D aspartic acid; E, glutamate, glutamic acid; F, phenylalanine; G, glycine; H histidine; I isoleucine; K, lysine; L, leucine; M, methionine; N, asparagine; P, proline; Q, glutamine; R, arginine; S, serine; T, threonine; V, valine; W, tryptophan; Y, tyrosine; Z, glutamine or glutamic acid. NAI-1540294631v3 38 [00107] The terms “peptide,” “protein,” and “polypeptide” are used interchangeably to refer to a natural or synthetic molecule comprising two or more amino acids linked by the carboxyl group of one amino acid to the alpha amino group of another. [00108] The terms “polypeptide fragment” or “fragment”, when used in reference to a particular polypeptide, refers to a polypeptide in which amino acid residues are deleted as compared to the reference polypeptide itself, but where the remaining amino acid sequence is usually identical to that of the reference polypeptide. Such deletions may occur at the amino-terminus or carboxy-terminus of the reference polypeptide, or alternatively both. Fragments typically are at least about 5, 6, 8 or 10 amino acids long, at least about 14 amino acids long, at least about 20, 30, 40 or 50 amino acids long, at least about 75 amino acids long, or at least about 100, 150, 200, 300, 500 or more amino acids long. A fragment can retain one or more of the biological activities of the reference polypeptide. In various embodiments, a fragment may comprise an enzymatic activity and/or an interaction site of the reference polypeptide. In another embodiment, a fragment may have immunogenic properties. [00109] As used herein, the term “substantially identical” or “substantially homologous” refers to a polypeptide or a nucleic acid molecule exhibiting at least about 50% identical or homologous to a reference amino acid sequence (for example, any of the amino acid sequences described herein) or a reference nucleotide sequence (for example, any of the nucleic acid sequences described herein). In certain embodiments, such a sequence is at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, or at least about 100% identical or homologous to the amino acid sequence or the nucleic acid sequence used for comparison. [00110] The term “identity” refers to sequence identity between two nucleic acid molecules or polypeptides. Identity can be determined by comparing a position in each sequence which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base, then the molecules are identical at that position. A degree of similarity or identity between nucleic acid or amino acid sequences is a function of the number of identical or matching nucleotides at positions shared by the nucleic acid sequences. Various alignment algorithms and/or programs may be used to calculate the identity between two sequences, including FASTA, or BLAST which are available as a part of the GCG sequence analysis package (University of Wisconsin, Madison, Wis.), and can be used with, e.g., default setting. When BLASTP is used, the percent similarity is based on the BLASTP positives score and the percent sequence identity is based on the BLASTP identities score. BLASTP “Identities” shows the number and fraction of total NAI-1540294631v3 39 residues in the high scoring sequence pairs which are identical; and BLASTP “Positives” shows the number and fraction of residues for which the alignment scores have positive values and which are similar to each other. Amino acid sequences having these degrees of identity or similarity or any intermediate degree of identity of similarity to the amino acid sequences disclosed herein are contemplated and encompassed by this disclosure. The polynucleotide sequences of similar polypeptides are deduced using the genetic code and may be obtained by conventional means, in particular by reverse translating its amino acid sequence using the genetic code. [00111] The term “nucleic acid” refers to a natural or synthetic molecule comprising a single nucleotide or two or more nucleotides linked by a phosphate group at the 3’ position of one nucleotide to the 5’ end of another nucleotide. The nucleic acid is not limited by length, and thus the nucleic acid can include deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). [00112] The term “operably linked to” refers to the functional relationship of a nucleic acid with another nucleic acid sequence. Promoters, enhancers, transcriptional and translational stop sites, and other signal sequences are examples of nucleic acid sequences operably linked to other sequences. For example, operable linkage of DNA to a transcriptional control element refers to the physical and functional relationship between the DNA and promoter such that the transcription of such DNA is initiated from the promoter by an RNA polymerase that specifically recognizes, binds to and transcribes the DNA. [00113] As used herein, the term “signal sequence” or “leader sequence” refers to a peptide sequence (e.g., 5, 10, 15, 20, 25 or 30 amino acids) present at the N-terminus of proteins that directs their entry to the secretory pathway. [00114] The term “variant” refers to an amino acid or peptide sequence having conservative amino acid substitutions, non-conservative amino acid substitutions (i.e. a degenerate variant), substitutions within the wobble position of each codon (i.e. DNA and RNA) encoding an amino acid, amino acids added to the C-terminus of a peptide, or a peptide having 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% sequence identity to a reference sequence. [00115] As used herein, the term “a conservative sequence modification” refers to an amino acid modification that does not significantly affect or alter the binding characteristics of the presently disclosed GPC3-targeted binding comprising the amino acid sequence. Conservative modifications can include amino acid substitutions, additions and deletions. Modifications can be introduced by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Amino acids can be classified into groups according to their physicochemical properties such as charge and polarity. Conservative amino acid substitutions are ones in which the NAI-1540294631v3 40 amino acid residue is replaced with an amino acid within the same group. For example, amino acids can be classified by charge: positively-charged amino acids include lysine, arginine, histidine, negatively-charged amino acids include aspartic acid, glutamic acid, neutral charge amino acids include alanine, asparagine, cysteine, glutamine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. In addition, amino acids can be classified by polarity: polar amino acids include arginine (basic polar), asparagine, aspartic acid (acidic polar), glutamic acid (acidic polar), glutamine, histidine (basic polar), lysine (basic polar), serine, threonine, and tyrosine; non-polar amino acids include alanine, cysteine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, and valine. Thus, one or more amino acid residues within a CDR region can be replaced with other amino acid residues from the same group and the altered antibody can be tested for retained function using the functional assays described herein. In certain embodiments, no more than one, no more than two, no more than three, no more than four, no more than five residues within a specified sequence or a CDR region are altered. [00116] The term “vector” refers to a nucleic acid sequence capable of transporting into a cell another nucleic acid to which the vector sequence has been linked. The term “expression vector” includes any vector, (e.g., a plasmid, cosmid or phage chromosome) containing a gene construct in a form suitable for expression by a cell (e.g., linked to a transcriptional control element). [00117] As used herein, the term “effective amount” refers to an amount sufficient to affect a beneficial or desired clinical result upon treatment. An effective amount can be administered to a subject in one or more doses. In certain embodiments, an effective amount can be an amount that is sufficient to palliate, ameliorate, stabilize, reverse or slow the progression of the disease, or otherwise reduce the pathological consequences of the disease. The effective amount can be determined by a physician on a case-by-case basis and is within the skill of one in the art. Several factors are typically taken into account when determining an appropriate dosage to achieve an effective amount. These factors include age, sex and weight of the subject, the condition being treated, the severity of the condition and the form and effective concentration of the cells administered. [00118] The term “pharmaceutically acceptable” refers to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit/risk ratio. NAI-1540294631v3 41 [00119] The term “treatment” refers to the medical management of a subject with the intent to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder. This term includes active treatment, that is, treatment directed specifically toward the improvement of a disease, pathological condition, or disorder, and also includes causal treatment, that is, treatment directed toward removal of the cause of the associated disease, pathological condition, or disorder. In addition, this term includes palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder. [00120] An “individual” or “subject” herein is a vertebrate, such as a human or a non-human animal, for example, a mammal. Mammals include, but are not limited to, humans, primates, farm animals, sport animals, rodents and pets. Non-limiting examples of non-human animal subjects include rodents such as mice, rats, hamsters, and guinea pigs, rabbits, dogs, cats, sheep, pigs, goats, cattle, horses, and non-human primates such as apes and monkeys. In certain embodiment, the subject is a mammal. In certain embodiment, the subject is a human. [00121] The term “GPC3 antigen” and, in particular, “GPC3” is intended to include fragments, variants (e.g., allelic variants), and derivatives of the antigen molecule, e.g., the GPC3 molecule. [00122] The terms “comprises”, “comprising”, and are intended to have the broad meaning ascribed to them in U.S. Patent Law and can mean “includes”, “including” and the like. 5.2. GPC3 [00123] GPC3 is also known as glypican-3. Glypican-3 (GPC3) is a 70 kDa protein that is anchored to the cell surface via glycosylphosphatidylinositol (GPI). GPC3 is widely expressed in the placenta and fetal tissues such as liver, lung, and kidney, and it regulates morphogenesis or growth. However, its expression is significantly reduced in adult organs. On the other hand, significantly high levels of GPC3 are expressed in hepatocellular carcinoma (HCC) cells compared to normal liver and non-neoplastic liver lesions, making GPC3 a promising tumor marker and a potential molecular target for therapeutic intervention in HCC. GPC3 is a member of the heparan sulfate (HS) proteoglycan family that consists of six members, GPC1 to GPC6, which serve as receptors or coreceptors for a number of ligand molecules, including morphogens, growth factors, adhesion, and matrix molecules, to regulate their signaling and distribution. NAI-1540294631v3 42 [00124] The presently disclosed molecule binds to a GPC3 antigen. In certain embodiments, the presently disclosed binding molecule binds to a human GPC3 antigen. In certain embodiments, the GPC3 is a human GPC3. In certain embodiments, the GPC3 is a wild-type human GPC3. In certain embodiments, the GPC3 antigen comprises or consists of the amino acid sequence with a Uniprot Reference No: P51654-1 (SEQ ID NO: 22), which is provided below. MAGTVRTACLVVAMLLSLDFPGQAQPPPPPPDATCHQVRSFFQRLQPGLKWVPETPVPGSDLQVCLP KGPTCCSRKMEEKYQLTARLNMEQLLQSASMELKFLIIQNAAVFQEAFEIVVRHAKNYTNAMFKNNY PSLTPQAFEFVGEFFTDVSLYILGSDINVDDMVNELFDSLFPVIYTQLMNPGLPDSALDINECLRGA RRDLKVFGNFPKLIMTQVSKSLQVTRIFLQALNLGIEVINTTDHLKFSKDCGRMLTRMWYCSYCQGL MMVKPCGGYCNVVMQGCMAGVVEIDKYWREYILSLEELVNGMYRIYDMENVLLGLFSTIHDSIQYVQ KNAGKLTTTIGKLCAHSQQRQYRSAYYPEDLFIDKKVLKVAHVEHEETLSSRRRELIQKLKSFISFY SALPGYICSHSPVAENDTLCWNGQELVERYSQKAARNGMKNQFNLHELKMKGPEPVVSQIIDKLKHI NQLLRTMSMPKGRVLDKNLDEEGFESGDCGDDEDECIGGSGDGMIKVKNQLRFLAELAYDLDVDDAP GNSQQATPKDNEISTFHNLGNVHSPLKLLTSMAISVVCFFFLVH [SEQ ID NO: 22] [00125] In certain embodiments, the GPC3 antigen comprises or consists of an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 100% identical to the amino acid sequence set forth in SEQ ID NO: 22 or a fragment thereof. [00126] In certain embodiments, the presently disclosed binding molecule binds to an epitope of GPC3. In certain embodiments, the epitope is within amino acids 524 to 563 of SEQ ID NO: 22. In certain embodiments, the epitope is at the C-terminus of GPC3. 5.3. Chimeric Antigen Receptors (CARs) [00127] The presently disclosed subject matter provides chimeric antigen receptors (CARs) that targets GPC3. [00128] CARs are engineered receptors, which graft or confer a specificity of interest onto an immune effector cell. CARs can be used to graft the specificity of a monoclonal antibody onto a cell, e.g., an immunoresponsive cell, e.g., a T cell or a NK cell; with transferring of their coding sequence, e.g., facilitated by retroviral vectors. [00129] In certain embodiments, the GPC3-targeted CAR comprises an extracellular domain that specifically binds to GPC3, a transmembrane domain, and an intracellular domain. In certain embodiments, the extracellular domain is fused to the transmembrane domain, which is fused to the intracellular domain. NAI-1540294631v3 43 [00130] In certain embodiments, the extracellular domain of the CAR comprises a GPC3-binding region, and is responsible for antigen recognition. In certain embodiments, the extracellular domain further comprise a signal peptide (SP) so that the CAR can be glycosylated and anchored in the cell membrane of a cell (e.g., an immunoresponsive cell). [00131] In certain embodiments, the transmembrane domain (TD) connects the extracellular domain to the intracellular domain and resides within the cell membrane when expressed by a cell (e.g., an immunoresponsive cell). [00132] In certain embodiments, the intracellular domain transmits an activation signal to the cell (e.g., an immunoresponsive cell) after antigen recognition. In certain embodiments, the intracellular domain comprises an intracellular signaling domain (ISD). In certain embodiments, the intracellular domain further comprises a co-stimulatory signaling region (CSR). A “signaling domain (SD)” generally comprises immunoreceptor tyrosine-based activation motifs (ITAMs) that activate a signaling cascade when the ITAM is phosphorylated. The term “co-stimulatory signaling region (CSR)” refers to an intracellular signaling domain from a costimulatory protein receptor, such as CD28, 41BB, and ICOS, that are able to enhance the activation of an immunoresponsive cell (e.g., a T cell). [00133] In certain embodiments, the disclosed CAR is defined by the formula: [00134] SP–ED-HG–TM–CSR–SD; or [00135] SP– ED–HG–TM–SD–CSR; wherein “SP” represents an optional signal peptide, wherein “ED” represents an extracellular domain that binds to GPC3. wherein “HG” represents an optional hinge domain, wherein “TM” represents a transmembrane domain, wherein “CSR” represents one or more co-stimulatory signaling regions, wherein “SD” represents a signaling domain, and wherein “–” represents a peptide bond or linker. [00136] Additional CAR constructs are described, for example, in Fresnak et al., Nat. Rev. Cancer 16(9):566-81 (2016), which is incorporated by reference in its entirety for the teaching of these CAR models. [00137] In certain embodiments, the CAR can be for example (and without limitation), a TRUCK, Universal CAR, a Self-driving CAR, an Armored CAR, a Self-destruct CAR, a Conditional CAR, a Marked CAR, a TenCAR, a Dual CAR, or a sCAR. NAI-1540294631v3 44 [00138] TRUCKs (T cells redirected for universal cytokine killing) co-express a chimeric antigen receptor (CAR) and an antitumor cytokine. Cytokine expression may be constitutive or induced by T cell activation. Targeted by CAR specificity, localized production of pro-inflammatory cytokines recruits endogenous immune cells to tumor sites and may potentiate an antitumor response. [00139] Universal, allogeneic CAR T cells are engineered to no longer express endogenous T cell receptor (TCR) and/or major histocompatibility complex (MHC) molecules, thereby preventing graft-versus-host disease (GVHD) or rejection, respectively. [00140] Self-driving CARs co-express a CAR and a chemokine receptor, which binds to a tumor ligand, thereby enhancing tumor homing. [00141] CAR T cells engineered to be resistant to immunosuppression (Armored CARs) may be genetically modified to no longer express various immune checkpoint molecules (e.g., cytotoxic T lymphocyte-associated antigen 4 (CTLA4) or programmed cell death protein 1 (PD-1)). Exemplary “Knockdown” and “Knockout” techniques include, but are not limited to, RNA interference (RNAi) (e.g., asRNA, miRNA, shRNA, siRNA, etc.) and CRISPR interference (CRISPRi) (e.g., CRISPR- Cas9). In certain embodiments, CAR T cells are engineered to express a dominant-negative form of a checkpoint molecule. In certain embodiments, the extracellular ligand-binding domain of the immune checkpoint molecule is fused to a transmembrane membrane in order to compete for ligand binding. For example, the extracellular ligand-binding domain of PD-1 may be fused to a CD8 transmembrane domain, thus competing for PD-1 ligand from the target cell. In certain embodiments, CAR T cells are engineered to express an immune checkpoint switch receptor to exploit the inhibitory immune checkpoint ligand present on a target cell. In certain embodiments, the extracellular ligand-binding domain of the immune checkpoint molecule is fused to a signaling, stimulatory, and/or co-stimulatory domain. For example, the extracellular ligand-binding domain of PD-1 may be fused to a CD28 domain, thus providing CD28 co-stimulation while blocking PD-1 signaling. In certain embodiments, the CAR T cells may be administered with an aptamer or a monoclonal antibody that blocks immune checkpoint signaling. In certain embodiments, the CAR T cells (e.g., CAR T cell therapy) are combined with a PD-1 blockade method, such as administration with PD-1/PD-L1 antagonistic aptamers or anti-PD-1/PD-L1 antibodies. In certain embodiments, the CAR T cells and PD-1 pathway-blocking antibodies are administered conjointly. In certain embodiments, the CAR T cells are engineered to express or express and secrete an immune checkpoint-blocking antibody, such as anti-PD-1 or anti-PD-L1, or fragments thereof. In certain embodiments, the CAR T cells are administered with a vector (e.g., an engineered virus) that expresses an immune checkpoint-blocking molecule described herein. NAI-1540294631v3 45 [00142] A self-destruct CAR may be designed using RNA delivered by electroporation to encode the CAR. Alternatively, inducible apoptosis of the T cell may be achieved based on ganciclovir binding to thymidine kinase in gene-modified lymphocytes or the more recently described system of activation of human caspase 9 by a small-molecule dimerizer. [00143] A conditional CAR T cell is by default unresponsive, or switched “off”, until the addition of a small molecule to complete the circuit, enabling full transduction of both signal 1 and signal 2, thereby activating the CAR T cell. Alternatively, T cells may be engineered to express an adaptor- specific receptor with affinity for subsequently administered secondary antibodies directed at target antigen. [00144] Marked CAR T cells express a CAR plus a tumor epitope to which an existing monoclonal antibody agent binds. In the setting of intolerable adverse effects, administration of the monoclonal antibody clears the CAR T cells and alleviates symptoms with no additional off-tumor effects. [00145] A tandem CAR (TanCAR) T cell expresses a single CAR consisting of two linked single- chain variable fragments (scFvs) that have different affinities fused to intracellular co-stimulatory domain(s) and a CD3ζ domain. TanCAR T cell activation is achieved only when target cells co- express both targets. [00146] A dual CAR T cell expresses two separate CARs with different ligand binding targets; one CAR includes only the CD3ζ domain and the other CAR includes only the co-stimulatory domain(s). Dual CAR T cell activation requires co-expression of both targets on the tumor. [00147] A safety CAR (sCAR) consists of an extracellular scFv fused to an intracellular inhibitory domain. sCAR T cells co-expressing a standard CAR become activated only when encountering target cells that possess the standard CAR target but lack the sCAR target. [00148] There are three generations of CARs. “First generation” CARs are typically composed of an extracellular antigen-binding domain (e.g., an scFv), which is fused to a transmembrane domain, which is fused to an intracellular domain. “First generation” CARs can provide de novo antigen recognition and cause activation of both CD4+ and CD8+ T-cells through their CD3ζ chain signaling domain in a single fusion molecule, independent of HLA-mediated antigen presentation. “Second generation” CARs add an intracellular signaling domain from a co-stimulatory molecule (e.g., CD28, 4-1BB, ICOS, OX40) to the intracellular domain of the CAR to provide additional signals to the cell. “Second generation” CARs comprise those that provide both co-stimulation (e.g., CD28 or 4-1BB) and activation (CD3ζ). “Third generation” CARs combine multiple signaling domains to further augment potency, e.g., are made by combining multiple signaling domains, such as CD3 ζ- NAI-1540294631v3 46 CD28-OC40 or CD3ζ-CD28-4-1BB, to augment potency with stronger cytokine production and killing ability. In certain embodiments, the GPC3-targeted CAR is a first-generation CAR. In certain embodiments, the GPC3-targeted CAR is a second-generation CAR. [00149] In certain embodiments, the CAR comprises one signaling domain. In certain embodiments, the CAR comprises one or more signaling domain (co-stimulatory signaling region). The one or more signaling domain may be a polypeptide selected from the group consisting of CD8, CD3ζ, CD3δ, CD3γ, CD3ε, FcγRI-γ, FcyRIII-y, FcεRIβ, FcεRIy, DAP10, DAP12, CD32, CD79a, CD79b, CD28, CD3C, CD4, B2C, 4-1BB (CD137), ICOS, CD27, CD288, CD80, NKp30, OX40, and mutants thereof. 5.3.1. Extracellular Domain of A CAR [00150] The extracellular domain of the CARs disclosed herein generally comprise an antigen recognition domain that binds a GPC3 antigen. Such antigen-specific binding domains are typically derived from an antibody. In certain embodiments, the extracellular domain of the CAR comprises or is a functional antibody fragment or derivative thereof (e.g., a single chain variable fragment (scFv) or a Fab, or any suitable antigen binding fragment of an antibody). [00151] In certain embodiments, the extracellular domain of the CAR comprises an scFv. In certain embodiments, the scFv is from a monoclonal antibody (mAb). The scFv can be a human scFv, a humanized scFv, or a murine scFv. In certain embodiments, the scFv is a human scFv. The scFv can be derived from fusing the heavy chain variable region (VH) and light chain variable region (VL) of an antibody. Alternatively or additionally, the scFv may be derived from Fab’s (instead of from an antibody, e.g., obtained from Fab libraries). [00152] In certain embodiments, the extracellular domain of the CAR comprises a Fab. In certain embodiments, the Fab is crosslinked. In certain embodiments, the extracellular domain of the CAR comprises a F(ab)2. [00153] Any of the foregoing molecules may be comprised in a fusion protein with a heterologous sequence to form the extracellular domain of the CAR. [00154] In certain embodiments, the extracellular domain of the CAR binds to a GPC3 antigen (e.g., a human GPC3 antigen) with a EC50 of between about 1 ng/mL and about 600 ng/mL, between about 1 ng/mL and about 500 ng/mL, between about 1 ng/mL and about 400 ng/mL, between about 1 ng/mL and about 300 ng/mL, between about 1 ng/mL and about 200 ng/mL, between about 1 ng/mL and about 150 ng/mL between about 1 ng/mL and about 100 ng/mL, between about 1 ng/mL and about 60 ng/mL, between about 1 ng/mL and about 50 ng/mL, between about 1 ng/mL and about 40 ng/mL, between about 4 ng/mL and about 60 ng/mL, between about 4 NAI-1540294631v3 47 ng/mL and about 50 ng/mL, or between about 4 ng/mL and about 40 ng/mL. In certain embodiments, the extracellular domain of the CAR binds to GPC3 antigen (e.g., a human GPC3 antigen) with a EC50 of between about 1 ng/mL and about 60 ng/mL. In certain embodiments, the extracellular domain of the CAR binds to GPC3 antigen (e.g., a human GPC3 antigen) with a EC50 of between about 4 ng/mL and about 40 ng/mL. [00155] Binding of the extracellular domain of the CAR can be confirmed by, for example, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), FACS analysis, bioassay (e.g., growth inhibition), or Western Blot assay. Each of these assays generally detect the presence of protein-antibody complexes of particular interest by employing a labeled reagent (e.g., an antibody, or a scFv) specific for the complex of interest. For example, the scFv can be radioactively labeled and used in a radioimmunoassay (RIA) (see, for example, Weintraub, B., Principles of Radioimmunoassays, Seventh Training Course on Radioligand Assay Techniques, The Endocrine Society, March, 1986, which is incorporated by reference herein). The radioactive isotope can be detected by such means as the use of a γ counter or a scintillation counter or by autoradiography. In certain embodiments, the GPC3-targeted extracellular domain is labeled with a fluorescent marker. Non-limiting examples of fluorescent markers include green fluorescent protein (GFP), blue fluorescent protein (e.g., EBFP, EBFP2, Azurite, and mKalama1), cyan fluorescent protein (e.g., ECFP, Cerulean, and CyPet), and yellow fluorescent protein (e.g., YFP, Citrine, Venus, and YPet). [00156] In certain embodiments, the extracellular domain of the CAR comprises a heavy chain variable region (VH) and a light chain variable region (VL). [00157] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 29; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 30. SEQ ID NO: 29 and SEQ ID NO: 30 are disclosed in Table 1. [00158] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25. SEQ ID NOs: 23-25 are provided in Table 1. NAI-1540294631v3 48 [00159] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 29. [00160] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28. SEQ ID NOs: 26-28 are provided in Table 1. [00161] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 30. [00162] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 NAI-1540294631v3 49 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28. [00163] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 29, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30. [00164] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00165] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 30. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31. SEQ ID NO: 31 is provided in Table 1. [00166] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 30. In certain NAI-1540294631v3 50 embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 32. SEQ ID NO: 32 is provided in Table 1. [00167] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 1 (ATA001 or ATA017) CDRs 1 2 3 VH DYAMH (SEQ ID NO: VISTYNGNTNYNRKFKD NDY (SEQ ID Q
Figure imgf000053_0001
[00168] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 29 is set forth in SEQ ID NO: 33, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACAAATTCACTGATTATGCTATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACTACAACCGGAAGTTTAAGGACAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAAG ATTCTGCCATCTATTACTGTTTAAAGAACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTC A (SEQ ID NO: 33) [00169] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 29 is set forth in SEQ ID NO: 34, which is provided below. CAAGTGCAGCTGCAGCAGTCTGGTCCTGAGGTGGTGCGGCCCGGAGTGTCCGTGAAAATCTCCTGCA AAGGCAGCGGCTACAAGTTCACAGACTACGCCATGCACTGGGTGAAGCAGAGCCACGCCAAGAGCCT GGAATGGATCGGCGTGATTTCTACATATAATGGCAATACCAACTACAACCGGAAGTTTAAGGACAAG NAI-1540294631v3 51 GCCACCATGACCGTGGACAAAAGCAGCAGCACCGCCTACATGGAACTGGCTAGACTGACCTCTGAGG ACAGCGCCATCTACTATTGTCTGAAGAACGACTACTGGGGCCAGGGCACAAGCGTCACCGTTTCCAG C (SEQ ID NO: 34) [00170] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 30 is set forth in SEQ ID NO: 35, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCTTCTCTT GCAAGTCAAGTCAGAGTCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGGTGTTACAGAGGCC AGGCCAGTCGCCAAATCGCCTAATCTATCTGGTGTCTAAATTGGACTCTAGAGTCCCTGACAGGTTC ACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACAGTTTCCTCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAA A (SEQ ID NO: 35) [00171] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 30 is set forth in SEQ ID NO: 36, which is provided below. GACGTGGTCATGACCCAGACCCCTCTGACACTGAGCGTGACCATCGGCCAGCCTGCTAGCTTCAGCT GCAAGAGCTCCCAGAGCCTGCTGGACTCTGATGGCAAGACCTACCTGAACTGGGTGCTGCAGAGACC TGGCCAATCTCCTAACAGACTGATCTACCTGGTGTCAAAGCTGGATTCTAGAGTGCCAGATAGATTC ACCGGCAGCGGCAGCGGAACCGATTTTACACTGAAGATCAGCCGGGTGGAAGCCGAGGACCTGGGAG TGTACTACTGCTGGCAGGGAACCCAGTTCCCCCACACATTCGGCGGCGGAACAAAGCTCGAGATCAA A (SEQ ID NO: 36) [00172] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 31 is set forth in SEQ ID NO: 37, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACAAATTCACTGATTATGCTATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACTACAACCGGAAGTTTAAGGACAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAAG ATTCTGCCATCTATTACTGTTTAAAGAACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTC AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACCCAGACT CCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCTTCTCTTGCAAGTCAAGTCAGAGTCTCT TAGATAGTGATGGAAAGACATATTTGAATTGGGTGTTACAGAGGCCAGGCCAGTCGCCAAATCGCCT AATCTATCTGGTGTCTAAATTGGACTCTAGAGTCCCTGACAGGTTCACTGGCAGTGGATCAGGGACA GATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGCAAGGTA CACAGTTTCCTCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA (SEQ ID NO: 37) NAI-1540294631v3 52 [00173] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 31 is set forth in SEQ ID NO: 38, which is provided below. CAAGTGCAGCTGCAGCAGTCTGGTCCTGAGGTGGTGCGGCCCGGAGTGTCCGTGAAAATCTCCTGCA AAGGCAGCGGCTACAAGTTCACAGACTACGCCATGCACTGGGTGAAGCAGAGCCACGCCAAGAGCCT GGAATGGATCGGCGTGATTTCTACATATAATGGCAATACCAACTACAACCGGAAGTTTAAGGACAAG GCCACCATGACCGTGGACAAAAGCAGCAGCACCGCCTACATGGAACTGGCTAGACTGACCTCTGAGG ACAGCGCCATCTACTATTGTCTGAAGAACGACTACTGGGGCCAGGGCACAAGCGTCACCGTTTCCAG CGGCGGCGGAGGCAGCGGCGGCGGCGGCTCTGGCGGCGGCGGAAGCGACGTGGTCATGACCCAGACC CCTCTGACACTGAGCGTGACCATCGGCCAGCCTGCTAGCTTCAGCTGCAAGAGCTCCCAGAGCCTGC TGGACTCTGATGGCAAGACCTACCTGAACTGGGTGCTGCAGAGACCTGGCCAATCTCCTAACAGACT GATCTACCTGGTGTCAAAGCTGGATTCTAGAGTGCCAGATAGATTCACCGGCAGCGGCAGCGGAACC GATTTTACACTGAAGATCAGCCGGGTGGAAGCCGAGGACCTGGGAGTGTACTACTGCTGGCAGGGAA CCCAGTTCCCCCACACATTCGGCGGCGGAACAAAGCTCGAGATCAAA (SEQ ID NO: 38) [00174] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 32 is set forth in SEQ ID NO: 39, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCTTCTCTT GCAAGTCAAGTCAGAGTCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGGTGTTACAGAGGCC AGGCCAGTCGCCAAATCGCCTAATCTATCTGGTGTCTAAATTGGACTCTAGAGTCCCTGACAGGTTC ACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACAGTTTCCTCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAA AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAGCTGCAGCAGTCT GGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACAAATTCA CTGATTATGCTATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATTAG TACTTACAATGGTAATACAAACTACAACCGGAAGTTTAAGGACAAGGCCACAATGACTGTAGACAAA TCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAAGATTCTGCCATCTATTACTGTT TAAAGAACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 39) [00175] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 32 is set forth in SEQ ID NO: 40, which is provided below. GACGTGGTCATGACCCAGACCCCTCTGACACTGAGCGTGACCATCGGCCAGCCTGCTAGCTTCAGCT GCAAGAGCTCCCAGAGCCTGCTGGACTCTGATGGCAAGACCTACCTGAACTGGGTGCTGCAGAGACC TGGCCAATCTCCTAACAGACTGATCTACCTGGTGTCAAAGCTGGATTCTAGAGTGCCAGATAGATTC ACCGGCAGCGGCAGCGGAACCGATTTTACACTGAAGATCAGCCGGGTGGAAGCCGAGGACCTGGGAG TGTACTACTGCTGGCAGGGAACCCAGTTCCCCCACACATTCGGCGGCGGAACAAAGCTCGAGATCAA NAI-1540294631v3 53 AGGCGGCGGAGGCAGCGGCGGCGGCGGCTCTGGCGGCGGCGGAAGCCAAGTGCAGCTGCAGCAGTCT GGTCCTGAGGTGGTGCGGCCCGGAGTGTCCGTGAAAATCTCCTGCAAAGGCAGCGGCTACAAGTTCA CAGACTACGCCATGCACTGGGTGAAGCAGAGCCACGCCAAGAGCCTGGAATGGATCGGCGTGATTTC TACATATAATGGCAATACCAACTACAACCGGAAGTTTAAGGACAAGGCCACCATGACCGTGGACAAA AGCAGCAGCACCGCCTACATGGAACTGGCTAGACTGACCTCTGAGGACAGCGCCATCTACTATTGTC TGAAGAACGACTACTGGGGCCAGGGCACAAGCGTCACCGTTTCCAGC (SEQ ID NO: 40) [00176] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 47; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 48. SEQ ID NO: 47 and SEQ ID NO: 48 are disclosed in Table 2. [00177] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43. SEQ ID NOs: 41-43 are provided in Table 2. [00178] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 47. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 47. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 47. [00179] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, NAI-1540294631v3 54 and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. SEQ ID NOs: 44-46 are provided in Table 2. [00180] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 48. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 48. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 48. [00181] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00182] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 47, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 48. [00183] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00184] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the NAI-1540294631v3 55 amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 47, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 48. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 49. SEQ ID NO: 49 is provided in Table 2. [00185] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 47, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 48. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 50. SEQ ID NO: 50 is provided in Table 2. [00186] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 2 (ATA002 or ATA018) CDRs 1 2 3 VI TYN NTNNN KFK TTY E ID
Figure imgf000058_0001
NAI-1540294631v3 56 VH-VL QVQLQQSGPEVVRPGVSVKISCKGSGYTFTDYAIHWVKQSHAKSLEWIGVISTY scFv NGNTNNNQKFKGKATMTVDKSSSTAYLELARLTSEDSAIYYCARSGTTYWGQGT A i TLTVSSGGGGSGGGGSGGGGSDVVMTQTPLTLSVTIGQPASISCKSSQSLLDSE
Figure imgf000059_0001
, sequence of SEQ ID NO: 47 is set forth in SEQ ID NO: 51, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTATGCTATACACTGGGTGAAGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACAACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGACATCTGAGG ATTCTGCCATCTATTACTGTGCAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGT CTCCTCA (SEQ ID NO: 51) [00188] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 47 is set forth in SEQ ID NO: 52, which is provided below. CAAGTGCAGCTGCAGCAGAGCGGACCTGAGGTGGTGCGGCCTGGCGTGTCTGTGAAGATCAGCTGCA AGGGCTCTGGCTACACCTTTACCGACTACGCCATTCACTGGGTCAAGCAGAGCCACGCCAAGTCTCT GGAGTGGATCGGCGTGATCTCCACCTATAACGGCAATACCAACAACAACCAGAAGTTCAAGGGCAAG GCCACCATGACCGTGGATAAGAGCAGCAGCACCGCCTACCTGGAACTTGCTAGACTCACAAGCGAGG ACAGCGCCATCTACTATTGCGCCAGAAGCGGAACAACCTACTGGGGCCAAGGCACCACCCTGACCGT GTCCAGC (SEQ ID NO: 52) [00189] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 48 is set forth in SEQ ID NO: 53, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGCCTCCTAGATAGTGAAGGAAAGACATATTTGAATTGGTTGTTACAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTC AGTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGTGGTGGGACCAAGCTGGAGCTGAA A (SEQ ID NO: 53) [00190] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 48 is set forth in SEQ ID NO: 54, which is provided below. NAI-1540294631v3 57 GATGTGGTCATGACACAGACACCTCTGACACTGAGCGTGACAATCGGCCAGCCTGCTAGCATCAGCT GTAAAAGCAGCCAGAGCCTGCTGGACTCTGAGGGCAAAACATACCTGAACTGGCTGCTGCAGAGGCC CGGCCAGAGTCCTAAGCGGCTGATCTACCTGGTGTCCAAGGTGGACAGCGGTGTTCCAGATAGATTC AGCGGAAGCGGCTCCGGCACCGACTTCACCCTGAAAATCTCTAGAGTGGAAGCCGAGGACCTGGGCG TGTACTACTGCTGGCAGGGCACCCACTTCCCCCATACCTTCGGCGGCGGCACAAAGCTGGAACTGAA G (SEQ ID NO: 54) [00191] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 49 is set forth in SEQ ID NO: 55, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTATGCTATACACTGGGTGAAGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACAACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGACATCTGAGG ATTCTGCCATCTATTACTGTGCAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGT CTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACC CAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAAGTCAGA GCCTCCTAGATAGTGAAGGAAAGACATATTTGAATTGGTTGTTACAGAGGCCAGGCCAGTCTCCAAA GCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTCAGTGGCAGTGGATCA GGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGC AAGGTACACATTTTCCTCACACGTTCGGTGGTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 55) [00192] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 49 is set forth in SEQ ID NO: 56, which is provided below CAAGTGCAGCTGCAGCAGAGCGGACCTGAGGTGGTGCGGCCTGGCGTGTCTGTGAAGATCAGCTGCA AGGGCTCTGGCTACACCTTTACCGACTACGCCATTCACTGGGTCAAGCAGAGCCACGCCAAGTCTCT GGAGTGGATCGGCGTGATCTCCACCTATAACGGCAATACCAACAACAACCAGAAGTTCAAGGGCAAG GCCACCATGACCGTGGATAAGAGCAGCAGCACCGCCTACCTGGAACTTGCTAGACTCACAAGCGAGG ACAGCGCCATCTACTATTGCGCCAGAAGCGGAACAACCTACTGGGGCCAAGGCACCACCCTGACCGT GTCCAGCGGCGGAGGAGGCAGCGGCGGCGGAGGCTCTGGCGGCGGCGGAAGTGATGTGGTCATGACA CAGACACCTCTGACACTGAGCGTGACAATCGGCCAGCCTGCTAGCATCAGCTGTAAAAGCAGCCAGA GCCTGCTGGACTCTGAGGGCAAAACATACCTGAACTGGCTGCTGCAGAGGCCCGGCCAGAGTCCTAA GCGGCTGATCTACCTGGTGTCCAAGGTGGACAGCGGTGTTCCAGATAGATTCAGCGGAAGCGGCTCC GGCACCGACTTCACCCTGAAAATCTCTAGAGTGGAAGCCGAGGACCTGGGCGTGTACTACTGCTGGC NAI-1540294631v3 58 AGGGCACCCACTTCCCCCATACCTTCGGCGGCGGCACAAAGCTGGAACTGAAG (SEQ ID NO: 56) [00193] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 50 is set forth in SEQ ID NO: 57, which is provided below GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGCCTCCTAGATAGTGAAGGAAAGACATATTTGAATTGGTTGTTACAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTC AGTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGTGGTGGGACCAAGCTGGAGCTGAA AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAGCTGCAGCAGTCT GGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACACATTCA CTGATTATGCTATACACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATTAG TACTTACAATGGTAATACAAACAACAACCAGAAGTTTAAGGGCAAGGCCACAATGACTGTAGACAAA TCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGACATCTGAGGATTCTGCCATCTATTACTGTG CAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA (SEQ ID NO: 57) [00194] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 50 is set forth in SEQ ID NO: 58, which is provided below. GATGTGGTCATGACACAGACACCTCTGACACTGAGCGTGACAATCGGCCAGCCTGCTAGCATCAGCT GTAAAAGCAGCCAGAGCCTGCTGGACTCTGAGGGCAAAACATACCTGAACTGGCTGCTGCAGAGGCC CGGCCAGAGTCCTAAGCGGCTGATCTACCTGGTGTCCAAGGTGGACAGCGGTGTTCCAGATAGATTC AGCGGAAGCGGCTCCGGCACCGACTTCACCCTGAAAATCTCTAGAGTGGAAGCCGAGGACCTGGGCG TGTACTACTGCTGGCAGGGCACCCACTTCCCCCATACCTTCGGCGGCGGCACAAAGCTGGAACTGAA GGGCGGAGGAGGCAGCGGCGGCGGAGGCTCTGGCGGCGGCGGAAGTCAAGTGCAGCTGCAGCAGAGC GGACCTGAGGTGGTGCGGCCTGGCGTGTCTGTGAAGATCAGCTGCAAGGGCTCTGGCTACACCTTTA CCGACTACGCCATTCACTGGGTCAAGCAGAGCCACGCCAAGTCTCTGGAGTGGATCGGCGTGATCTC CACCTATAACGGCAATACCAACAACAACCAGAAGTTCAAGGGCAAGGCCACCATGACCGTGGATAAG AGCAGCAGCACCGCCTACCTGGAACTTGCTAGACTCACAAGCGAGGACAGCGCCATCTACTATTGCG CCAGAAGCGGAACAACCTACTGGGGCCAAGGCACCACCCTGACCGTGTCCAGC (SEQ ID NO: 58) [00195] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 62; and/or ii) the VL comprises NAI-1540294631v3 59 a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 63. SEQ ID NO: 62 and SEQ ID NO: 63 are disclosed in Table 3. [00196] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61. SEQ ID NOs: 59-61 are provided in Table 3. [00197] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 62. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 62. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 62. [00198] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. SEQ ID NOs: 26, 27, and 46 are provided in Table 3. [00199] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 63. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous NAI-1540294631v3 60 to the amino acid sequence set forth in SEQ ID NO: 63. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 63. [00200] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00201] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 62, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 63. [00202] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00203] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 62, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 63. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid NAI-1540294631v3 61 sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 64. SEQ ID NO: 64 is provided in Table 3. [00204] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 62, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 63. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 65. SEQ ID NO: 65 is provided in Table 3. [00205] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 3 (ATA003) CDRs 1 2 3 VH DYAVH (SEQ ID NO: VISTYNGNTNYNQKFKG SGTSY (SEQ ID Q
Figure imgf000064_0001
NAI-1540294631v3 62 [00206] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 62 is set forth in SEQ ID NO: 66, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTATGCTGTGCACTGGGTGAAACAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACTACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAGG ATTCTGCCATCTATTACTGTGCAAGATCGGGGACGAGCTACTGGGGCCAAGGCACCACTCTCACAGT CTCCTCA (SEQ ID NO: 66) [00207] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 62 is set forth in SEQ ID NO: 67, which is provided below. CAAGTGCAGCTGCAGCAGAGCGGTCCTGAGGTGGTGCGGCCTGGAGTGTCCGTGAAGATTAGCTGCA AGGGCAGCGGATATACATTTACCGACTACGCCGTGCACTGGGTTAAGCAGAGCCACGCCAAGTCTCT GGAATGGATCGGCGTGATCAGCACCTACAACGGCAACACCAACTACAATCAGAAATTCAAGGGCAAG GCCACCATGACCGTGGACAAGAGCAGCAGCACCGCCTACATGGAACTGGCCAGGCTGACCAGCGAGG ATTCCGCCATCTACTACTGCGCCAGAAGCGGAACCTCATATTGGGGCCAGGGCACTACACTGACAGT GTCCAGC (SEQ ID NO: 67) [00208] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 63 is set forth in SEQ ID NO: 68, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGTTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGTCTCTTAGATAGTGATGGAAAGACCTATTTGAATTGGTTGTTACAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTTTCTGGTGTCTAAACTGGACTCTGGAGCCCCTGACAGGTTC TCTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGTGCTGGGACCAAGCTGGAGCTGAA A (SEQ ID NO: 68) [00209] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 63 is set forth in SEQ ID NO: 69, which is provided below GATGTGGTCATGACACAGACCCCTCTGACCCTTTCTTTTACCATCGGACAGCCTGCTAGCATCAGCT GCAAGAGCTCTCAGAGCCTGCTGGACAGCGACGGCAAAACCTACCTGAACTGGCTGCTGCAAAGACC CGGCCAGTCTCCAAAGCGGCTGATCTTCCTGGTGTCCAAGCTGGACAGCGGCGCTCCTGATAGATTC AGCGGCAGCGGAAGCGGCACAGACTTCACACTGAAAATCTCTAGAGTGGAAGCCGAGGACCTGGGCG TGTACTACTGTTGGCAGGGCACCCATTTCCCCCACACCTTCGGCGCCGGCACAAAGCTCGAGCTGAA G (SEQ ID NO: 69) NAI-1540294631v3 63 [00210] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 64 is set forth in SEQ ID NO: 70, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTATGCTGTGCACTGGGTGAAACAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACTACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAGG ATTCTGCCATCTATTACTGTGCAAGATCGGGGACGAGCTACTGGGGCCAAGGCACCACTCTCACAGT CTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACC CAGACTCCACTCACTTTGTCGTTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAAGTCAGA GTCTCTTAGATAGTGATGGAAAGACCTATTTGAATTGGTTGTTACAGAGGCCAGGCCAGTCTCCAAA GCGCCTAATCTTTCTGGTGTCTAAACTGGACTCTGGAGCCCCTGACAGGTTCTCTGGCAGTGGATCA GGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGC AAGGTACACATTTTCCTCACACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 70) [00211] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 64 is set forth in SEQ ID NO: 71, which is provided below CAAGTGCAGCTGCAGCAGAGCGGTCCTGAGGTGGTGCGGCCTGGAGTGTCCGTGAAGATTAGCTGCA AGGGCAGCGGATATACATTTACCGACTACGCCGTGCACTGGGTTAAGCAGAGCCACGCCAAGTCTCT GGAATGGATCGGCGTGATCAGCACCTACAACGGCAACACCAACTACAATCAGAAATTCAAGGGCAAG GCCACCATGACCGTGGACAAGAGCAGCAGCACCGCCTACATGGAACTGGCCAGGCTGACCAGCGAGG ATTCCGCCATCTACTACTGCGCCAGAAGCGGAACCTCATATTGGGGCCAGGGCACTACACTGACAGT GTCCAGCGGCGGCGGAGGCAGCGGCGGCGGCGGCTCCGGCGGAGGCGGCTCTGATGTGGTCATGACA CAGACCCCTCTGACCCTTTCTTTTACCATCGGACAGCCTGCTAGCATCAGCTGCAAGAGCTCTCAGA GCCTGCTGGACAGCGACGGCAAAACCTACCTGAACTGGCTGCTGCAAAGACCCGGCCAGTCTCCAAA GCGGCTGATCTTCCTGGTGTCCAAGCTGGACAGCGGCGCTCCTGATAGATTCAGCGGCAGCGGAAGC GGCACAGACTTCACACTGAAAATCTCTAGAGTGGAAGCCGAGGACCTGGGCGTGTACTACTGTTGGC AGGGCACCCATTTCCCCCACACCTTCGGCGCCGGCACAAAGCTCGAGCTGAAG (SEQ ID NO: 71) [00212] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 65 is set forth in SEQ ID NO: 72, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGTTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGTCTCTTAGATAGTGATGGAAAGACCTATTTGAATTGGTTGTTACAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTTTCTGGTGTCTAAACTGGACTCTGGAGCCCCTGACAGGTTC NAI-1540294631v3 64 TCTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGTGCTGGGACCAAGCTGGAGCTGAA AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAGCTGCAGCAGTCT GGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACACATTCA CTGATTATGCTGTGCACTGGGTGAAACAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATTAG TACTTACAATGGTAATACAAACTACAACCAGAAGTTTAAGGGCAAGGCCACAATGACTGTAGACAAA TCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAGGATTCTGCCATCTATTACTGTG CAAGATCGGGGACGAGCTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA (SEQ ID NO: 72) [00213] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 65 is set forth in SEQ ID NO: 73, which is provided below. GATGTGGTCATGACACAGACCCCTCTGACCCTTTCTTTTACCATCGGACAGCCTGCTAGCATCAGCT GCAAGAGCTCTCAGAGCCTGCTGGACAGCGACGGCAAAACCTACCTGAACTGGCTGCTGCAAAGACC CGGCCAGTCTCCAAAGCGGCTGATCTTCCTGGTGTCCAAGCTGGACAGCGGCGCTCCTGATAGATTC AGCGGCAGCGGAAGCGGCACAGACTTCACACTGAAAATCTCTAGAGTGGAAGCCGAGGACCTGGGCG TGTACTACTGTTGGCAGGGCACCCATTTCCCCCACACCTTCGGCGCCGGCACAAAGCTCGAGCTGAA GGGCGGCGGAGGCAGCGGCGGCGGCGGCTCCGGCGGAGGCGGCTCTCAAGTGCAGCTGCAGCAGAGC GGTCCTGAGGTGGTGCGGCCTGGAGTGTCCGTGAAGATTAGCTGCAAGGGCAGCGGATATACATTTA CCGACTACGCCGTGCACTGGGTTAAGCAGAGCCACGCCAAGTCTCTGGAATGGATCGGCGTGATCAG CACCTACAACGGCAACACCAACTACAATCAGAAATTCAAGGGCAAGGCCACCATGACCGTGGACAAG AGCAGCAGCACCGCCTACATGGAACTGGCCAGGCTGACCAGCGAGGATTCCGCCATCTACTACTGCG CCAGAAGCGGAACCTCATATTGGGGCCAGGGCACTACACTGACAGTGTCCAGC (SEQ ID NO: 73) [00214] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 75; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 76. SEQ ID NO: 75 and SEQ ID NO: 76 are disclosed in Table 4. [00215] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, NAI-1540294631v3 65 and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25. SEQ ID NOs: 23, 60, and 25 are provided in Table 4. [00216] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 75. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 75. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 75. [00217] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. SEQ ID NOs: 74, 45, and 46 are provided in Table 4. [00218] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 76. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 76. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 76. [00219] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification, and a CDR3 comprising the amino acid sequence set NAI-1540294631v3 66 forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00220] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 75, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76. [00221] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00222] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 76. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 77. SEQ ID NO: 77 is provided in Table 4. [00223] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain NAI-1540294631v3 67 embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 76. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 78. SEQ ID NO: 78 is provided in Table 4. [00224] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 4 (ATA004 or ATA019) CDRs 1 2 3 VH DYAMH (SEQ ID NO: VISTYNGNTNYNQKFKG NDY (SEQ ID Q N V L E N V L C L E K I
Figure imgf000070_0001
[00225] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 75 is set forth in SEQ ID NO: 79, which is provided below CAGGTCCAACTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTATGCTATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATCAGTACTTACAATGGTAATACAAACTACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAGG ATTCTGCCATCTATTACTGTCTAAAGAACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTC A (SEQ ID NO: 79) [00226] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 75 is set forth in SEQ ID NO: 80, which is provided below NAI-1540294631v3 68 CAGGTGCAGCTGCAGCAGAGCGGTCCTGAGGTGGTTAGACCTGGCGTGTCTGTGAAGATTAGCTGCA AAGGCAGCGGCTACACCTTTACCGACTACGCCATGCATTGGGTCAAGCAGAGCCACGCCAAGTCTCT GGAATGGATCGGCGTGATCTCTACATATAATGGCAACACCAACTACAACCAGAAGTTCAAGGGCAAG GCCACCATGACCGTGGATAAGAGCTCCAGCACCGCCTACATGGAACTGGCTAGACTGACAAGCGAGG ATAGCGCCATCTACTACTGTCTGAAGAACGACTACTGGGGCCAGGGCACCAGCGTGACCGTGAGCAG C (SEQ ID NO: 80) [00227] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 76 is set forth in SEQ ID NO: 81, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGCCTCTTAAATAGTGATGGAAAGACATATTTGAATTGGTTGTTACAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTC ACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAAGATTTGGGAG TTTTTTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAA A (SEQ ID NO: 81) [00228] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 76 is set forth in SEQ ID NO: 82, which is provided below. GACGTGGTGATGACACAGACCCCTCTGACCCTGAGCGTGACAATCGGCCAACCTGCTAGCATCTCTT GCAAGTCCTCCCAGAGCCTGCTGAACAGCGACGGCAAAACCTACCTGAATTGGCTGCTCCAGCGGCC TGGACAATCTCCAAAGCGGCTGATCTACCTGGTGAGCAAGGTGGACAGCGGCGTCCCCGATAGATTT ACCGGCAGCGGAAGCGGCACAGACTTCACTCTGAAAATCTCTAGAGTGGAAGCCGAGGACCTGGGAG TGTTCTACTGCTGGCAGGGAACCCACTTCCCCCACACATTCGGCGGCGGAACAAAGCTGGAGATCAA G (SEQ ID NO: 82) [00229] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 77 is set forth in SEQ ID NO: 83, which is provided below. CAGGTCCAACTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTATGCTATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATCAGTACTTACAATGGTAATACAAACTACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAGG ATTCTGCCATCTATTACTGTCTAAAGAACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTC AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACCCAGACT CCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAAGTCAGAGCCTCT TAAATAGTGATGGAAAGACATATTTGAATTGGTTGTTACAGAGGCCAGGCCAGTCTCCAAAGCGCCT AATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTCACTGGCAGTGGATCAGGGACA NAI-1540294631v3 69 GATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAAGATTTGGGAGTTTTTTATTGCTGGCAAGGTA CACATTTTCCTCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA (SEQ ID NO: 83) [00230] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 77 is set forth in SEQ ID NO: 84, which is provided below CAGGTGCAGCTGCAGCAGAGCGGTCCTGAGGTGGTTAGACCTGGCGTGTCTGTGAAGATTAGCTGCA AAGGCAGCGGCTACACCTTTACCGACTACGCCATGCATTGGGTCAAGCAGAGCCACGCCAAGTCTCT GGAATGGATCGGCGTGATCTCTACATATAATGGCAACACCAACTACAACCAGAAGTTCAAGGGCAAG GCCACCATGACCGTGGATAAGAGCTCCAGCACCGCCTACATGGAACTGGCTAGACTGACAAGCGAGG ATAGCGCCATCTACTACTGTCTGAAGAACGACTACTGGGGCCAGGGCACCAGCGTGACCGTGAGCAG CGGCGGCGGAGGCTCTGGAGGCGGAGGCAGCGGCGGCGGCGGCTCCGACGTGGTGATGACACAGACC CCTCTGACCCTGAGCGTGACAATCGGCCAACCTGCTAGCATCTCTTGCAAGTCCTCCCAGAGCCTGC TGAACAGCGACGGCAAAACCTACCTGAATTGGCTGCTCCAGCGGCCTGGACAATCTCCAAAGCGGCT GATCTACCTGGTGAGCAAGGTGGACAGCGGCGTCCCCGATAGATTTACCGGCAGCGGAAGCGGCACA GACTTCACTCTGAAAATCTCTAGAGTGGAAGCCGAGGACCTGGGAGTGTTCTACTGCTGGCAGGGAA CCCACTTCCCCCACACATTCGGCGGCGGAACAAAGCTGGAGATCAAG (SEQ ID NO: 84) [00231] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 78 is set forth in SEQ ID NO: 85, which is provided below GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGCCTCTTAAATAGTGATGGAAAGACATATTTGAATTGGTTGTTACAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTC ACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAAGATTTGGGAG TTTTTTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAA AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAACTGCAGCAGTCT GGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACACATTCA CTGATTATGCTATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATCAG TACTTACAATGGTAATACAAACTACAACCAGAAGTTTAAGGGCAAGGCCACAATGACTGTAGACAAA TCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAGGATTCTGCCATCTATTACTGTC TAAAGAACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 85) [00232] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 78 is set forth in SEQ ID NO: 86, which is provided below. GACGTGGTGATGACACAGACCCCTCTGACCCTGAGCGTGACAATCGGCCAACCTGCTAGCATCTCTT GCAAGTCCTCCCAGAGCCTGCTGAACAGCGACGGCAAAACCTACCTGAATTGGCTGCTCCAGCGGCC TGGACAATCTCCAAAGCGGCTGATCTACCTGGTGAGCAAGGTGGACAGCGGCGTCCCCGATAGATTT NAI-1540294631v3 70 ACCGGCAGCGGAAGCGGCACAGACTTCACTCTGAAAATCTCTAGAGTGGAAGCCGAGGACCTGGGAG TGTTCTACTGCTGGCAGGGAACCCACTTCCCCCACACATTCGGCGGCGGAACAAAGCTGGAGATCAA GGGCGGCGGAGGCTCTGGAGGCGGAGGCAGCGGCGGCGGCGGCTCCCAGGTGCAGCTGCAGCAGAGC GGTCCTGAGGTGGTTAGACCTGGCGTGTCTGTGAAGATTAGCTGCAAAGGCAGCGGCTACACCTTTA CCGACTACGCCATGCATTGGGTCAAGCAGAGCCACGCCAAGTCTCTGGAATGGATCGGCGTGATCTC TACATATAATGGCAACACCAACTACAACCAGAAGTTCAAGGGCAAGGCCACCATGACCGTGGATAAG AGCTCCAGCACCGCCTACATGGAACTGGCTAGACTGACAAGCGAGGATAGCGCCATCTACTACTGTC TGAAGAACGACTACTGGGGCCAGGGCACCAGCGTGACCGTGAGCAGC (SEQ ID NO: 86) [00233] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 89; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 90. SEQ ID NO: 89 and SEQ ID NO: 90 are disclosed in Table 5. [00234] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43. SEQ ID NOs: 87, 88, and 43 are provided in Table 5. [00235] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 89. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 89. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 89. [00236] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set NAI-1540294631v3 71 forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. SEQ ID NOs: 26, 27, and 46 are provided in Table 5. [00237] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 90. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 90. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 90. [00238] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00239] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 89, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 90. [00240] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00241] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a NAI-1540294631v3 72 CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 89, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 90. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 91. SEQ ID NO: 91 is provided in Table 5. [00242] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 89, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 90. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 92. SEQ ID NO: 92 is provided in Table 5. [00243] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 5 (ATA005) CDRs 1 2 3 D Q N L L E
Figure imgf000075_0001
NAI-1540294631v3 73 VH-VL QVQLQQSGPEVERPGVSVKISCKGSGYTFTDFAIHWVIQSHAKSLEWIGVISTYN scFv GHTNYNQKFKGKATMTVDKSSSTAYLELARLISEDSGIYYCARSGTTYWGQGTTL A i TVSSGGGGSGGGGSGGGGSDVVMTQTPLTLSVTIGQPASISCKSSQSLLDSDGKT Y L E I I
Figure imgf000076_0001
, sequence of SEQ ID NO: 89 is set forth in SEQ ID NO: 93, which is provided below. CAGGTCCAACTGCAGCAGTCTGGGCCTGAGGTGGAGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTTTGCTATACACTGGGTGATTCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTCATACAAACTACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGATATCTGAGG ATTCTGGCATCTATTACTGTGCAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGT CTCCTCC (SEQ ID NO: 93) [00245] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 89 is set forth in SEQ ID NO: 94, which is provided below CAGGTGCAGCTGCAGCAGTCCGGCCCCGAGGTGGAAAGACCTGGCGTTAGCGTGAAGATCAGCTGCA AGGGCTCTGGCTACACATTCACAGACTTTGCCATCCACTGGGTGATCCAGAGCCACGCCAAGAGCCT GGAATGGATCGGCGTGATCAGCACCTATAACGGCCACACCAACTACAACCAGAAGTTCAAGGGAAAA GCTACAATGACCGTGGACAAGTCCTCTTCTACCGCCTACCTCGAGCTGGCCAGACTGATTAGCGAGG ACAGCGGAATCTACTATTGCGCCCGGAGCGGCACCACCTACTGGGGCCAGGGCACAACCCTGACCGT GTCCAGC (SEQ ID NO: 94) [00246] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 90 is set forth in SEQ ID NO: 95, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGCCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGTTGTTCCAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTATCTAGTGTCTAAATTGGACTCTGGAGTCCCTGACAGGTTC ACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGTGCTGGGACCAAGCTGGAGCTGAA A (SEQ ID NO: 95) [00247] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 90 is set forth in SEQ ID NO: 96, which is provided below. NAI-1540294631v3 74 GATGTGGTGATGACACAGACCCCTCTGACCCTGAGCGTGACAATCGGACAACCTGCTAGCATCAGCT GTAAAAGCTCTCAGAGCCTGCTGGACAGCGACGGCAAGACCTACCTGAATTGGCTGTTCCAGAGACC AGGGCAAAGCCCCAAGCGGCTGATCTACCTGGTGTCCAAGCTGGATTCTGGCGTGCCTGACAGGTTC ACCGGCAGCGGATCAGGCACAGATTTCACCCTTAAAATCAGCAGAGTGGAGGCCGAGGACCTGGGCG TCTACTACTGCTGGCAGGGCACACATTTCCCTCACACCTTTGGCGCCGGCACCAAGCTGGAACTGAA G (SEQ ID NO: 96) [00248] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 91 is set forth in SEQ ID NO: 97, which is provided below. CAGGTCCAACTGCAGCAGTCTGGGCCTGAGGTGGAGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTTTGCTATACACTGGGTGATTCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTCATACAAACTACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGATATCTGAGG ATTCTGGCATCTATTACTGTGCAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGT CTCCTCCGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACC CAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAAGTCAGA GCCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGTTGTTCCAGAGGCCAGGCCAGTCTCCAAA GCGCCTAATCTATCTAGTGTCTAAATTGGACTCTGGAGTCCCTGACAGGTTCACTGGCAGTGGATCA GGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGC AAGGTACACATTTTCCTCACACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 97) [00249] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 91 is set forth in SEQ ID NO: 98, which is provided below. CAGGTGCAGCTGCAGCAGTCCGGCCCCGAGGTGGAAAGACCTGGCGTTAGCGTGAAGATCAGCTGCA AGGGCTCTGGCTACACATTCACAGACTTTGCCATCCACTGGGTGATCCAGAGCCACGCCAAGAGCCT GGAATGGATCGGCGTGATCAGCACCTATAACGGCCACACCAACTACAACCAGAAGTTCAAGGGAAAA GCTACAATGACCGTGGACAAGTCCTCTTCTACCGCCTACCTCGAGCTGGCCAGACTGATTAGCGAGG ACAGCGGAATCTACTATTGCGCCCGGAGCGGCACCACCTACTGGGGCCAGGGCACAACCCTGACCGT GTCCAGCGGCGGCGGCGGCAGCGGCGGCGGAGGTTCTGGAGGAGGCGGCAGCGATGTGGTGATGACA CAGACCCCTCTGACCCTGAGCGTGACAATCGGACAACCTGCTAGCATCAGCTGTAAAAGCTCTCAGA GCCTGCTGGACAGCGACGGCAAGACCTACCTGAATTGGCTGTTCCAGAGACCAGGGCAAAGCCCCAA GCGGCTGATCTACCTGGTGTCCAAGCTGGATTCTGGCGTGCCTGACAGGTTCACCGGCAGCGGATCA GGCACAGATTTCACCCTTAAAATCAGCAGAGTGGAGGCCGAGGACCTGGGCGTCTACTACTGCTGGC NAI-1540294631v3 75 AGGGCACACATTTCCCTCACACCTTTGGCGCCGGCACCAAGCTGGAACTGAAG (SEQ ID NO: 98) [00250] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 92 is set forth in SEQ ID NO: 99, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGCCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGTTGTTCCAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTATCTAGTGTCTAAATTGGACTCTGGAGTCCCTGACAGGTTC ACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGTGCTGGGACCAAGCTGGAGCTGAA AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAACTGCAGCAGTCT GGGCCTGAGGTGGAGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACACATTCA CTGATTTTGCTATACACTGGGTGATTCAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATTAG TACTTACAATGGTCATACAAACTACAACCAGAAGTTTAAGGGCAAGGCCACAATGACTGTAGACAAA TCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGATATCTGAGGATTCTGGCATCTATTACTGTG CAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCC (SEQ ID NO: 99) [00251] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 92 is set forth in SEQ ID NO: 100, which is provided below. GATGTGGTGATGACACAGACCCCTCTGACCCTGAGCGTGACAATCGGACAACCTGCTAGCATCAGCT GTAAAAGCTCTCAGAGCCTGCTGGACAGCGACGGCAAGACCTACCTGAATTGGCTGTTCCAGAGACC AGGGCAAAGCCCCAAGCGGCTGATCTACCTGGTGTCCAAGCTGGATTCTGGCGTGCCTGACAGGTTC ACCGGCAGCGGATCAGGCACAGATTTCACCCTTAAAATCAGCAGAGTGGAGGCCGAGGACCTGGGCG TCTACTACTGCTGGCAGGGCACACATTTCCCTCACACCTTTGGCGCCGGCACCAAGCTGGAACTGAA GGGCGGCGGCGGCAGCGGCGGCGGAGGTTCTGGAGGAGGCGGCAGCCAGGTGCAGCTGCAGCAGTCC GGCCCCGAGGTGGAAAGACCTGGCGTTAGCGTGAAGATCAGCTGCAAGGGCTCTGGCTACACATTCA CAGACTTTGCCATCCACTGGGTGATCCAGAGCCACGCCAAGAGCCTGGAATGGATCGGCGTGATCAG CACCTATAACGGCCACACCAACTACAACCAGAAGTTCAAGGGAAAAGCTACAATGACCGTGGACAAG TCCTCTTCTACCGCCTACCTCGAGCTGGCCAGACTGATTAGCGAGGACAGCGGAATCTACTATTGCG CCCGGAGCGGCACCACCTACTGGGGCCAGGGCACAACCCTGACCGTGTCCAGC (SEQ ID NO: 100) [00252] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 101; and/or ii) the VL NAI-1540294631v3 76 comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 102. SEQ ID NO: 101 and SEQ ID NO: 102 are disclosed in Table 6. [00253] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43. SEQ ID NOs: 41-43 are provided in Table 6. [00254] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 101. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 101. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 101. [00255] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. SEQ ID NOs: 44-46 are provided in Table 6. [00256] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 102. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or NAI-1540294631v3 77 homologous to the amino acid sequence set forth in SEQ ID NO: 102. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 102. [00257] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00258] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 101, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 102. [00259] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00260] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 101, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 102. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid NAI-1540294631v3 78 sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 103. SEQ ID NO: 103 is provided in Table 6. [00261] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 101, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 102. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 104. SEQ ID NO: 104 is provided in Table 6. [00262] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 6 (ATA006) CDRs 1 2 3 VH DYAIH (SEQ ID NO: VISTYNGNTNNNQKFKG SGTTY (SEQ ID Q N L L E N L T Y L E M I
Figure imgf000081_0001
NAI-1540294631v3 79 [00263] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 101 is set forth in SEQ ID NO: 105, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTATGCTATACACTGGGTGATGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACAACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGACATCTGAGG ATTCTGCCATCTATTACTGTGCAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGT CTCCTCA (SEQ ID NO: 105) [00264] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 101 is set forth in SEQ ID NO: 106, which is provided below. CAAGTGCAGCTGCAGCAGAGCGGCCCTGAGGTGGTGCGGCCCGGAGTGTCCGTGAAGATCAGCTGCA AGGGCTCTGGCTACACCTTTACCGACTACGCCATCCACTGGGTGATGCAGAGCCATGCCAAGAGCCT GGAGTGGATCGGAGTGATCTCTACATACAACGGCAACACCAACAACAATCAGAAGTTCAAGGGAAAA GCCACAATGACCGTGGATAAGTCCTCCAGCACCGCCTACCTGGAACTGGCTAGACTGACTAGCGAGG ATAGCGCTATTTACTATTGCGCCAGAAGCGGCACCACATACTGGGGCCAGGGAACAACACTGACCGT GTCAAGC (SEQ ID NO: 106) [00265] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 102 is set forth in SEQ ID NO: 107, which is provided below. GATGTTGTGATGACCCAGTCTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGCCTCCTAGATAGTGAAGGAAAGACATATCTGAATTGGTTGTTACAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTC AGTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGTGGTGGGACCAAGCTGGAGCTGAA A (SEQ ID NO: 107) [00266] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 102 is set forth in SEQ ID NO: 108, which is provided below. GACGTGGTCATGACCCAGAGTCCTCTGACCCTGAGCGTCACAATCGGCCAGCCTGCCAGCATCAGCT GTAAAAGCAGCCAGAGCCTGCTGGACTCTGAGGGCAAAACCTACCTGAACTGGCTGCTGCAAAGACC TGGACAGTCTCCAAAGCGGCTGATCTACCTGGTTTCCAAGGTGGACTCTGGCGTGCCTGATAGATTC AGCGGCTCCGGCAGCGGCACCGACTTCACCCTGAAGATCAGCCGCGTGGAAGCCGAGGACCTGGGAG TGTACTACTGCTGGCAGGGCACCCACTTCCCCCACACATTCGGCGGCGGTACCAAGCTGGAACTCAA G (SEQ ID NO: 108) NAI-1540294631v3 80 [00267] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 103 is set forth in SEQ ID NO: 109, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGATTATGCTATACACTGGGTGATGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACAACAACCAGAAGTTTAAGGGCAAG GCCACAATGACTGTAGACAAATCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGACATCTGAGG ATTCTGCCATCTATTACTGTGCAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGT CTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACC CAGTCTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAAGTCAGA GCCTCCTAGATAGTGAAGGAAAGACATATCTGAATTGGTTGTTACAGAGGCCAGGCCAGTCTCCAAA GCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTCAGTGGCAGTGGATCA GGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGC AAGGTACACATTTTCCTCACACGTTCGGTGGTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 109) [00268] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 103 is set forth in SEQ ID NO: 110, which is provided below. CAAGTGCAGCTGCAGCAGAGCGGCCCTGAGGTGGTGCGGCCCGGAGTGTCCGTGAAGATCAGCTGCA AGGGCTCTGGCTACACCTTTACCGACTACGCCATCCACTGGGTGATGCAGAGCCATGCCAAGAGCCT GGAGTGGATCGGAGTGATCTCTACATACAACGGCAACACCAACAACAATCAGAAGTTCAAGGGAAAA GCCACAATGACCGTGGATAAGTCCTCCAGCACCGCCTACCTGGAACTGGCTAGACTGACTAGCGAGG ATAGCGCTATTTACTATTGCGCCAGAAGCGGCACCACATACTGGGGCCAGGGAACAACACTGACCGT GTCAAGCGGCGGCGGCGGCAGCGGCGGAGGCGGCTCTGGCGGAGGCGGCAGCGACGTGGTCATGACC CAGAGTCCTCTGACCCTGAGCGTCACAATCGGCCAGCCTGCCAGCATCAGCTGTAAAAGCAGCCAGA GCCTGCTGGACTCTGAGGGCAAAACCTACCTGAACTGGCTGCTGCAAAGACCTGGACAGTCTCCAAA GCGGCTGATCTACCTGGTTTCCAAGGTGGACTCTGGCGTGCCTGATAGATTCAGCGGCTCCGGCAGC GGCACCGACTTCACCCTGAAGATCAGCCGCGTGGAAGCCGAGGACCTGGGAGTGTACTACTGCTGGC AGGGCACCCACTTCCCCCACACATTCGGCGGCGGTACCAAGCTGGAACTCAAG (SEQ ID NO: 110) [00269] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 104 is set forth in SEQ ID NO: 111, which is provided below. GATGTTGTGATGACCCAGTCTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTT GCAAGTCAAGTCAGAGCCTCCTAGATAGTGAAGGAAAGACATATCTGAATTGGTTGTTACAGAGGCC AGGCCAGTCTCCAAAGCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTC NAI-1540294631v3 81 AGTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGTACACATTTTCCTCACACGTTCGGTGGTGGGACCAAGCTGGAGCTGAA AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAGCTGCAGCAGTCT GGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACACATTCA CTGATTATGCTATACACTGGGTGATGCAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATTAG TACTTACAATGGTAATACAAACAACAACCAGAAGTTTAAGGGCAAGGCCACAATGACTGTAGACAAA TCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGACATCTGAGGATTCTGCCATCTATTACTGTG CAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA (SEQ ID NO: 111) [00270] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 104 is set forth in SEQ ID NO: 112, which is provided below. GACGTGGTCATGACCCAGAGTCCTCTGACCCTGAGCGTCACAATCGGCCAGCCTGCCAGCATCAGCT GTAAAAGCAGCCAGAGCCTGCTGGACTCTGAGGGCAAAACCTACCTGAACTGGCTGCTGCAAAGACC TGGACAGTCTCCAAAGCGGCTGATCTACCTGGTTTCCAAGGTGGACTCTGGCGTGCCTGATAGATTC AGCGGCTCCGGCAGCGGCACCGACTTCACCCTGAAGATCAGCCGCGTGGAAGCCGAGGACCTGGGAG TGTACTACTGCTGGCAGGGCACCCACTTCCCCCACACATTCGGCGGCGGTACCAAGCTGGAACTCAA GGGCGGCGGCGGCAGCGGCGGAGGCGGCTCTGGCGGAGGCGGCAGCCAAGTGCAGCTGCAGCAGAGC GGCCCTGAGGTGGTGCGGCCCGGAGTGTCCGTGAAGATCAGCTGCAAGGGCTCTGGCTACACCTTTA CCGACTACGCCATCCACTGGGTGATGCAGAGCCATGCCAAGAGCCTGGAGTGGATCGGAGTGATCTC TACATACAACGGCAACACCAACAACAATCAGAAGTTCAAGGGAAAAGCCACAATGACCGTGGATAAG TCCTCCAGCACCGCCTACCTGGAACTGGCTAGACTGACTAGCGAGGATAGCGCTATTTACTATTGCG CCAGAAGCGGCACCACATACTGGGGCCAGGGAACAACACTGACCGTGTCAAGC (SEQ ID NO: 112) [00271] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 119; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 120. SEQ ID NO: 119 and SEQ ID NO: 120 are disclosed in Table 7. [00272] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: NAI-1540294631v3 82 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115. SEQ ID NOs: 113-115 are provided in Table 7. [00273] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 119. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 119. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 119. [00274] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. SEQ ID NOs: 116-118 are provided in Table 7. [00275] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 120. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 120. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 120. [00276] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification, and a CDR3 comprising the amino acid sequence NAI-1540294631v3 83 set forth in SEQ ID NO: 118 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. [00277] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 119, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 120. [00278] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00279] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 120. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 121. SEQ ID NO: 121 is provided in Table 7. [00280] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set NAI-1540294631v3 84 forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 120. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 122. SEQ ID NO: 122 is provided in Table 7. [00281] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 7 (ATA007) CDRs 1 2 3 VH TYGVH (SEQ ID NO: VIWSGGSTDYNASFIS RGNDGPPFAY Q G Q D G Q Y Y D G K R
Figure imgf000087_0001
[00282] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 119 is set forth in SEQ ID NO: 123, which is provided below. CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTCTCATTAACTACCTATGGTGTTCACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGAGTGGCTGGGAGTGATATGGAGTGGTGGAAGCACAGACTATAATGCATCTTTCATATCCAGACTG ACCATCAGCAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAAGCTAATGACA CAGCCATATATTACTGTGCCAGAAGAGGGAATGATGGTCCTCCGTTTGCTTACTGGGGCCAAGGGAC TCTGGTCACTGTCTCTGCA (SEQ ID NO: 123) NAI-1540294631v3 85 [00283] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 119 is set forth in SEQ ID NO: 124, which is provided below. CAAGTGCAGCTGAAGCAGAGCGGACCTGGCCTGGTCCAGCCTAGCCAAAGCCTGAGCATCACATGCA CCGTGTCCGGCTTCAGCCTGACAACATACGGCGTGCACTGGGTGCGGCAGAGCCCCGGCAAGGGCCT GGAATGGCTGGGCGTTATCTGGAGCGGCGGATCTACCGACTACAACGCCAGCTTTATCTCTAGACTG ACCATCAGCAAGGACAACTCTAAAAGCCAGGTGTTCTTCAAGATGAACAGCCTTCAGGCCAACGACA CCGCCATCTACTATTGTGCCAGGCGGGGCAATGACGGCCCTCCTTTCGCCTACTGGGGCCAGGGCAC CCTGGTGACCGTGTCCGCC (SEQ ID NO: 124) [00284] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 120 is set forth in SEQ ID NO: 125, which is provided below. GACATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTT GTCGGGCAAGTCAGGAAATTAGTGGTTACTTAAGCTGGCTTCAGCAGAAACCAGATGGAACTATTAA ACGCCTGATCTACGCCGCATCCACTTTAGATTCTGGTGTCCCAAAAAGGTTCAGTGGCAGTAGGTCT GGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGCAGACTATTACTGTCTAC AATATGTTAGTTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 125) [00285] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 120 is set forth in SEQ ID NO: 126, which is provided below. GATATCCAGATGACCCAGTCTCCCAGCAGCCTGAGCGCTTCTCTCGGCGAGAGAGTGTCCCTGACCT GCAGAGCCAGCCAGGAGATCAGCGGCTACCTGTCCTGGCTGCAGCAGAAGCCTGATGGCACAATTAA GCGGCTGATCTATGCCGCTAGCACCCTGGACAGCGGCGTGCCAAAGAGATTTAGCGGAAGCAGAAGC GGCAGCGACTACAGCCTGACCATCAGCTCTCTGGAAAGCGAGGACTTCGCTGATTACTACTGCCTGC AGTACGTGTCCTACCCCCTGACATTCGGCGCCGGAACAAAGCTGGAGCTGAAA (SEQ ID NO: 126) [00286] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 121 is set forth in SEQ ID NO: 127, which is provided below. CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTCTCATTAACTACCTATGGTGTTCACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGAGTGGCTGGGAGTGATATGGAGTGGTGGAAGCACAGACTATAATGCATCTTTCATATCCAGACTG ACCATCAGCAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAAGCTAATGACA CAGCCATATATTACTGTGCCAGAAGAGGGAATGATGGTCCTCCGTTTGCTTACTGGGGCCAAGGGAC TCTGGTCACTGTCTCTGCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGAC ATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTC NAI-1540294631v3 86 GGGCAAGTCAGGAAATTAGTGGTTACTTAAGCTGGCTTCAGCAGAAACCAGATGGAACTATTAAACG CCTGATCTACGCCGCATCCACTTTAGATTCTGGTGTCCCAAAAAGGTTCAGTGGCAGTAGGTCTGGG TCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGCAGACTATTACTGTCTACAAT ATGTTAGTTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 127) [00287] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 121 is set forth in SEQ ID NO: 128, which is provided below. CAAGTGCAGCTGAAGCAGAGCGGACCTGGCCTGGTCCAGCCTAGCCAAAGCCTGAGCATCACATGCA CCGTGTCCGGCTTCAGCCTGACAACATACGGCGTGCACTGGGTGCGGCAGAGCCCCGGCAAGGGCCT GGAATGGCTGGGCGTTATCTGGAGCGGCGGATCTACCGACTACAACGCCAGCTTTATCTCTAGACTG ACCATCAGCAAGGACAACTCTAAAAGCCAGGTGTTCTTCAAGATGAACAGCCTTCAGGCCAACGACA CCGCCATCTACTATTGTGCCAGGCGGGGCAATGACGGCCCTCCTTTCGCCTACTGGGGCCAGGGCAC CCTGGTGACCGTGTCCGCCGGCGGAGGCGGATCTGGCGGCGGCGGTTCAGGCGGAGGCGGCTCTGAT ATCCAGATGACCCAGTCTCCCAGCAGCCTGAGCGCTTCTCTCGGCGAGAGAGTGTCCCTGACCTGCA GAGCCAGCCAGGAGATCAGCGGCTACCTGTCCTGGCTGCAGCAGAAGCCTGATGGCACAATTAAGCG GCTGATCTATGCCGCTAGCACCCTGGACAGCGGCGTGCCAAAGAGATTTAGCGGAAGCAGAAGCGGC AGCGACTACAGCCTGACCATCAGCTCTCTGGAAAGCGAGGACTTCGCTGATTACTACTGCCTGCAGT ACGTGTCCTACCCCCTGACATTCGGCGCCGGAACAAAGCTGGAGCTGAAA (SEQ ID NO: 128) [00288] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 122 is set forth in SEQ ID NO: 129, which is provided below. GACATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTT GTCGGGCAAGTCAGGAAATTAGTGGTTACTTAAGCTGGCTTCAGCAGAAACCAGATGGAACTATTAA ACGCCTGATCTACGCCGCATCCACTTTAGATTCTGGTGTCCCAAAAAGGTTCAGTGGCAGTAGGTCT GGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGCAGACTATTACTGTCTAC AATATGTTAGTTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAAGGTGGAGGTGGATC AGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTG CAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTACCTATGGTGTTC ACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAGCAC AGACTATAATGCATCTTTCATATCCAGACTGACCATCAGCAAGGACAATTCCAAGAGCCAAGTTTTC TTTAAAATGAACAGTCTGCAAGCTAATGACACAGCCATATATTACTGTGCCAGAAGAGGGAATGATG GTCCTCCGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 129) [00289] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 122 is set forth in SEQ ID NO: 130, which is provided below. NAI-1540294631v3 87 GATATCCAGATGACCCAGTCTCCCAGCAGCCTGAGCGCTTCTCTCGGCGAGAGAGTGTCCCTGACCT GCAGAGCCAGCCAGGAGATCAGCGGCTACCTGTCCTGGCTGCAGCAGAAGCCTGATGGCACAATTAA GCGGCTGATCTATGCCGCTAGCACCCTGGACAGCGGCGTGCCAAAGAGATTTAGCGGAAGCAGAAGC GGCAGCGACTACAGCCTGACCATCAGCTCTCTGGAAAGCGAGGACTTCGCTGATTACTACTGCCTGC AGTACGTGTCCTACCCCCTGACATTCGGCGCCGGAACAAAGCTGGAGCTGAAAGGCGGAGGCGGATC TGGCGGCGGCGGTTCAGGCGGAGGCGGCTCTCAAGTGCAGCTGAAGCAGAGCGGACCTGGCCTGGTC CAGCCTAGCCAAAGCCTGAGCATCACATGCACCGTGTCCGGCTTCAGCCTGACAACATACGGCGTGC ACTGGGTGCGGCAGAGCCCCGGCAAGGGCCTGGAATGGCTGGGCGTTATCTGGAGCGGCGGATCTAC CGACTACAACGCCAGCTTTATCTCTAGACTGACCATCAGCAAGGACAACTCTAAAAGCCAGGTGTTC TTCAAGATGAACAGCCTTCAGGCCAACGACACCGCCATCTACTATTGTGCCAGGCGGGGCAATGACG GCCCTCCTTTCGCCTACTGGGGCCAGGGCACCCTGGTGACCGTGTCCGCC (SEQ ID NO: 130) [00290] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 137; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 138. SEQ ID NO: 137 and SEQ ID NO: 138 are disclosed in Table 8. [00291] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133. SEQ ID NOs: 131-133 are provided in Table 8. [00292] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 137. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 137. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 137. [00293] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino NAI-1540294631v3 88 acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136. SEQ ID NOs: 134-136 are provided in Table 8. [00294] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 138. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 138. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 138. [00295] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136. [00296] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 137, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138. [00297] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of NAI-1540294631v3 89 the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00298] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 138. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 139. SEQ ID NO: 139 is provided in Table 8. [00299] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 138. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 140. SEQ ID NO: 140 is provided in Table 8. [00300] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 8 (ATA008 or ATA020) CDRs 1 2 3 D
Figure imgf000092_0001
N - 5 0 9 63 v3 90 VL RASGNIHNYLA (SEQ NAKTLAD (SEQ ID NO: QHFWTTPFT (SEQ ID NO: 134) 135) ID NO: 136) V LK SGPGLV PS SLSITCTVSGFSLNSYGVHWVR SPGKGLEWLGAIWSSG M A G M G D A G K N
Figure imgf000093_0001
sequence of SEQ ID NO: 137 is set forth in SEQ ID NO: 141, which is provided below. CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTCTCATTAAATAGCTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGAGTGGCTGGGAGCGATATGGAGTAGTGGAAGCACAGACTATAATGCACCTTTCATATCCAGACTG AGCATCAGCAAGGACAACTCCAAGAACCAAGTTTTCTTTAAAATGAACAGTCTGCAAGTTGATGACA CAGCCATATATTTTTGTGCCGGAAATCCAGAATCGGATCATTACTACGGCTACGAGGCCATGGACTC CTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 141) [00302] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 137 is set forth in SEQ ID NO: 142, which is provided below. CAAGTGCAGCTGAAGCAGAGCGGTCCTGGCCTGGTTCAGCCATCTCAGAGCCTGAGCATCACATGCA CCGTGTCCGGCTTTAGCCTGAACAGCTATGGCGTGCATTGGGTGCGGCAGTCTCCTGGCAAAGGCCT GGAATGGCTGGGCGCCATTTGGTCCAGCGGCTCCACCGACTACAACGCCCCTTTCATCTCTAGACTG AGCATCAGCAAGGACAACAGCAAGAACCAGGTCTTTTTCAAGATGAACAGCCTGCAAGTGGACGACA CCGCCATCTACTTCTGCGCCGGCAACCCCGAGAGCGACCACTACTACGGCTACGAGGCCATGGACAG CTGGGGCCAGGGCACCAGCGTGACAGTGTCCAGC (SEQ ID NO: 142) [00303] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 138 is set forth in SEQ ID NO: 143, which is provided below. GACATCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACAT GTCGAGCAAGTGGGAATATTCACAATTATTTAGCATGGTATCAGCAGAGACAGGGAAAATCTCCTCA NAI-1540294631v3 91 GCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGTGGCAGTGGATCA GGAACACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAAGATTTTGGGAGTTATTACTGTCAAC ATTTTTGGACTACTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAA (SEQ ID NO: 143) [00304] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 138 is set forth in SEQ ID NO: 144, which is provided below. GATATCCAGATGACACAGAGCCCTGCTTCTCTCTCCGCCAGCGTGGGCGAGACAGTGACCATCACCT GTAGAGCCAGCGGAAATATCCACAACTACCTGGCCTGGTACCAGCAGCGGCAAGGCAAGAGCCCCCA GCTGCTGGTGTACAATGCCAAGACCCTGGCTGATGGCGTGCCCAGCAGATTCAGCGGATCCGGCTCT GGCACACAGTACAGCCTGAAAATCATCAGCCTGCAGCCTGAGGATTTCGGAAGCTACTATTGCCAGC ACTTCTGGACCACCCCTTTCACCTTCGGCTCTGGCACTAAGCTGGAAATCAAG (SEQ ID NO: 144) [00305] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 139 is set forth in SEQ ID NO: 145, which is provided below. CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTCTCATTAAATAGCTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGAGTGGCTGGGAGCGATATGGAGTAGTGGAAGCACAGACTATAATGCACCTTTCATATCCAGACTG AGCATCAGCAAGGACAACTCCAAGAACCAAGTTTTCTTTAAAATGAACAGTCTGCAAGTTGATGACA CAGCCATATATTTTTGTGCCGGAAATCCAGAATCGGATCATTACTACGGCTACGAGGCCATGGACTC CTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGT GGAGGTGGATCTGACATCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTG TCACCATCACATGTCGAGCAAGTGGGAATATTCACAATTATTTAGCATGGTATCAGCAGAGACAGGG AAAATCTCCTCAGCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGT GGCAGTGGATCAGGAACACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAAGATTTTGGGAGTT ATTACTGTCAACATTTTTGGACTACTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAA (SEQ ID NO: 145) [00306] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 139 is set forth in SEQ ID NO: 146, which is provided below. CAAGTGCAGCTGAAGCAGAGCGGTCCTGGCCTGGTTCAGCCATCTCAGAGCCTGAGCATCACATGCA CCGTGTCCGGCTTTAGCCTGAACAGCTATGGCGTGCATTGGGTGCGGCAGTCTCCTGGCAAAGGCCT GGAATGGCTGGGCGCCATTTGGTCCAGCGGCTCCACCGACTACAACGCCCCTTTCATCTCTAGACTG AGCATCAGCAAGGACAACAGCAAGAACCAGGTCTTTTTCAAGATGAACAGCCTGCAAGTGGACGACA CCGCCATCTACTTCTGCGCCGGCAACCCCGAGAGCGACCACTACTACGGCTACGAGGCCATGGACAG NAI-1540294631v3 92 CTGGGGCCAGGGCACCAGCGTGACAGTGTCCAGCGGCGGAGGCGGATCTGGCGGAGGCGGCTCTGGA GGCGGCGGCTCAGATATCCAGATGACACAGAGCCCTGCTTCTCTCTCCGCCAGCGTGGGCGAGACAG TGACCATCACCTGTAGAGCCAGCGGAAATATCCACAACTACCTGGCCTGGTACCAGCAGCGGCAAGG CAAGAGCCCCCAGCTGCTGGTGTACAATGCCAAGACCCTGGCTGATGGCGTGCCCAGCAGATTCAGC GGATCCGGCTCTGGCACACAGTACAGCCTGAAAATCATCAGCCTGCAGCCTGAGGATTTCGGAAGCT ACTATTGCCAGCACTTCTGGACCACCCCTTTCACCTTCGGCTCTGGCACTAAGCTGGAAATCAAG (SEQ ID NO: 146) [00307] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 140 is set forth in SEQ ID NO: 147, which is provided below. GACATCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACAT GTCGAGCAAGTGGGAATATTCACAATTATTTAGCATGGTATCAGCAGAGACAGGGAAAATCTCCTCA GCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGTGGCAGTGGATCA GGAACACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAAGATTTTGGGAGTTATTACTGTCAAC ATTTTTGGACTACTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAAGGTGGAGGTGGATC AGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTG CAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAAATAGCTATGGTGTAC ACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGCGATATGGAGTAGTGGAAGCAC AGACTATAATGCACCTTTCATATCCAGACTGAGCATCAGCAAGGACAACTCCAAGAACCAAGTTTTC TTTAAAATGAACAGTCTGCAAGTTGATGACACAGCCATATATTTTTGTGCCGGAAATCCAGAATCGG ATCATTACTACGGCTACGAGGCCATGGACTCCTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 147) [00308] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 140 is set forth in SEQ ID NO: 148, which is provided below. GATATCCAGATGACACAGAGCCCTGCTTCTCTCTCCGCCAGCGTGGGCGAGACAGTGACCATCACCT GTAGAGCCAGCGGAAATATCCACAACTACCTGGCCTGGTACCAGCAGCGGCAAGGCAAGAGCCCCCA GCTGCTGGTGTACAATGCCAAGACCCTGGCTGATGGCGTGCCCAGCAGATTCAGCGGATCCGGCTCT GGCACACAGTACAGCCTGAAAATCATCAGCCTGCAGCCTGAGGATTTCGGAAGCTACTATTGCCAGC ACTTCTGGACCACCCCTTTCACCTTCGGCTCTGGCACTAAGCTGGAAATCAAGGGCGGAGGCGGATC TGGCGGAGGCGGCTCTGGAGGCGGCGGCTCACAAGTGCAGCTGAAGCAGAGCGGTCCTGGCCTGGTT CAGCCATCTCAGAGCCTGAGCATCACATGCACCGTGTCCGGCTTTAGCCTGAACAGCTATGGCGTGC ATTGGGTGCGGCAGTCTCCTGGCAAAGGCCTGGAATGGCTGGGCGCCATTTGGTCCAGCGGCTCCAC CGACTACAACGCCCCTTTCATCTCTAGACTGAGCATCAGCAAGGACAACAGCAAGAACCAGGTCTTT TTCAAGATGAACAGCCTGCAAGTGGACGACACCGCCATCTACTTCTGCGCCGGCAACCCCGAGAGCG NAI-1540294631v3 93 ACCACTACTACGGCTACGAGGCCATGGACAGCTGGGGCCAGGGCACCAGCGTGACAGTGTCCAGC (SEQ ID NO: 148) [00309] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 155; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 156. SEQ ID NO: 155 and SEQ ID NO: 156 are disclosed in Table 9. [00310] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151. SEQ ID NOs: 149-151 are provided in Table 9. [00311] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 155. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 155. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 155. [00312] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154. SEQ ID NOs: 152-154 are provided in Table 9. [00313] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 156. For example, the VL comprises an amino NAI-1540294631v3 94 acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 156. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 156. [00314] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154. [00315] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 155, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 156. [00316] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00317] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154. In certain NAI-1540294631v3 95 embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 156. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 157. SEQ ID NO: 157 is provided in Table 9. [00318] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 156. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 158. SEQ ID NO: 158 is provided in Table 9. [00319] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 9 (ATA009 or ATA021) CDRs 1 2 3 V DAYIH (SEQ ID NO: GIDPENDNTEYDPKFPG RPLGLPFDY : : N Q F L N Q S E
Figure imgf000098_0001
NAI-1540294631v3 96 VL-VH DIVLSQSPSSLAVSVGENVTMNCKSSQRLLDSSNQRNYLAWYQQKPGQSPQLLIF scFv WASTRESGVPDRFTGSGSGTDFTLTIRSVKAEDLAVYYCQQYYSYPFTFGSGTKL A i EIKGGGGSGGGGSGGGGSEVQLQQSGAEHVKPGTSVRLSCTASGFNIKDAYIHWV A
Figure imgf000099_0001
sequence of SEQ ID NO: 155 is set forth in SEQ ID NO: 159, which is provided below. GAGGTTCAGCTTCAGCAGTCTGGGGCAGAGCATGTGAAGCCAGGGACCTCAGTCAGGTTGTCCTGCA CAGCTTCTGGCTTTAACATTAAAGACGCCTATATACACTGGGTGAAGCAGAGGCCTGAACAGGGCCT GGAGTGGATTGGAGGGATTGATCCTGAGAATGACAATACTGAATATGACCCTAAATTCCCCGGCAAG GCCACTGTTGTGGCCGACACATCCTCCAACACAGCCTACCTGCAACTCAGCAGCCTGACATCTGAGG ACACTGCCGTCTATTACTGTGCTAGAAGACCTCTCGGACTACCTTTTGACTACTGGGGCCAAGGCAC CACTCTCACAGTCTCCTCA (SEQ ID NO: 159) [00321] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 155 is set forth in SEQ ID NO: 160, which is provided below. GAAGTGCAGCTTCAACAATCTGGCGCCGAGCACGTGAAGCCTGGCACAAGCGTGCGGCTGAGCTGCA CCGCCAGCGGATTTAACATCAAGGACGCCTATATCCACTGGGTCAAGCAGAGACCTGAGCAGGGACT GGAATGGATCGGCGGCATCGACCCCGAGAACGACAACACCGAGTACGACCCTAAGTTCCCTGGCAAG GCCACCGTTGTGGCCGATACATCTTCTAATACCGCCTACCTGCAGCTGTCCTCCCTGACCAGCGAGG ACACCGCTGTGTACTATTGTGCCAGACGGCCTCTGGGCCTGCCTTTCGACTACTGGGGCCAGGGCAC AACCCTCACAGTGTCCAGC (SEQ ID NO: 160) [00322] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 156 is set forth in SEQ ID NO: 161, which is provided below. GACATTGTGCTGTCACAGTCTCCATCCTCCCTAGCTGTGTCAGTTGGAGAGAACGTTACTATGAACT GCAAGTCCAGTCAGAGACTTTTAGATAGTAGCAATCAAAGGAACTATTTGGCCTGGTACCAGCAGAA ACCAGGGCAGTCTCCTCAACTGCTGATTTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGC TTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGAAGTGTGAAGGCTGAAGACCTGG CAGTTTATTACTGTCAGCAATATTATAGCTATCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAAT AAAA (SEQ ID NO: 161) [00323] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 156 is set forth in SEQ ID NO: 162, which is provided below. GACATCGTGCTGAGCCAGAGCCCTAGCAGCCTGGCCGTGTCTGTGGGCGAGAATGTGACAATGAACT GCAAGAGCTCTCAGAGGCTGCTGGACAGCAGCAACCAGCGGAACTACCTGGCTTGGTACCAGCAAAA GCCCGGCCAGAGCCCCCAGCTGCTGATTTTCTGGGCCTCCACCAGAGAGAGCGGCGTGCCAGATAGA NAI-1540294631v3 97 TTCACCGGCAGCGGCAGCGGCACTGATTTCACCCTGACCATCAGAAGCGTGAAAGCTGAAGATCTGG CCGTGTACTACTGCCAGCAGTACTACAGCTACCCCTTTACATTCGGCAGCGGCACCAAGCTGGAAAT CAAA (SEQ ID NO: 162) [00324] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 157 is set forth in SEQ ID NO: 163, which is provided below. GAGGTTCAGCTTCAGCAGTCTGGGGCAGAGCATGTGAAGCCAGGGACCTCAGTCAGGTTGTCCTGCA CAGCTTCTGGCTTTAACATTAAAGACGCCTATATACACTGGGTGAAGCAGAGGCCTGAACAGGGCCT GGAGTGGATTGGAGGGATTGATCCTGAGAATGACAATACTGAATATGACCCTAAATTCCCCGGCAAG GCCACTGTTGTGGCCGACACATCCTCCAACACAGCCTACCTGCAACTCAGCAGCCTGACATCTGAGG ACACTGCCGTCTATTACTGTGCTAGAAGACCTCTCGGACTACCTTTTGACTACTGGGGCCAAGGCAC CACTCTCACAGTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGAC ATTGTGCTGTCACAGTCTCCATCCTCCCTAGCTGTGTCAGTTGGAGAGAACGTTACTATGAACTGCA AGTCCAGTCAGAGACTTTTAGATAGTAGCAATCAAAGGAACTATTTGGCCTGGTACCAGCAGAAACC AGGGCAGTCTCCTCAACTGCTGATTTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTC ACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGAAGTGTGAAGGCTGAAGACCTGGCAG TTTATTACTGTCAGCAATATTATAGCTATCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAA A (SEQ ID NO: 163) [00325] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 157 is set forth in SEQ ID NO: 164, which is provided below. GAAGTGCAGCTTCAACAATCTGGCGCCGAGCACGTGAAGCCTGGCACAAGCGTGCGGCTGAGCTGCA CCGCCAGCGGATTTAACATCAAGGACGCCTATATCCACTGGGTCAAGCAGAGACCTGAGCAGGGACT GGAATGGATCGGCGGCATCGACCCCGAGAACGACAACACCGAGTACGACCCTAAGTTCCCTGGCAAG GCCACCGTTGTGGCCGATACATCTTCTAATACCGCCTACCTGCAGCTGTCCTCCCTGACCAGCGAGG ACACCGCTGTGTACTATTGTGCCAGACGGCCTCTGGGCCTGCCTTTCGACTACTGGGGCCAGGGCAC AACCCTCACAGTGTCCAGCGGAGGCGGCGGATCCGGCGGTGGCGGATCTGGCGGAGGCGGCAGCGAC ATCGTGCTGAGCCAGAGCCCTAGCAGCCTGGCCGTGTCTGTGGGCGAGAATGTGACAATGAACTGCA AGAGCTCTCAGAGGCTGCTGGACAGCAGCAACCAGCGGAACTACCTGGCTTGGTACCAGCAAAAGCC CGGCCAGAGCCCCCAGCTGCTGATTTTCTGGGCCTCCACCAGAGAGAGCGGCGTGCCAGATAGATTC ACCGGCAGCGGCAGCGGCACTGATTTCACCCTGACCATCAGAAGCGTGAAAGCTGAAGATCTGGCCG TGTACTACTGCCAGCAGTACTACAGCTACCCCTTTACATTCGGCAGCGGCACCAAGCTGGAAATCAA A (SEQ ID NO: 164) [00326] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 158 is set forth in SEQ ID NO: 165, which is provided below. NAI-1540294631v3 98 GACATTGTGCTGTCACAGTCTCCATCCTCCCTAGCTGTGTCAGTTGGAGAGAACGTTACTATGAACT GCAAGTCCAGTCAGAGACTTTTAGATAGTAGCAATCAAAGGAACTATTTGGCCTGGTACCAGCAGAA ACCAGGGCAGTCTCCTCAACTGCTGATTTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGC TTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGAAGTGTGAAGGCTGAAGACCTGG CAGTTTATTACTGTCAGCAATATTATAGCTATCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAAT AAAAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGAGGTTCAGCTTCAGCAG TCTGGGGCAGAGCATGTGAAGCCAGGGACCTCAGTCAGGTTGTCCTGCACAGCTTCTGGCTTTAACA TTAAAGACGCCTATATACACTGGGTGAAGCAGAGGCCTGAACAGGGCCTGGAGTGGATTGGAGGGAT TGATCCTGAGAATGACAATACTGAATATGACCCTAAATTCCCCGGCAAGGCCACTGTTGTGGCCGAC ACATCCTCCAACACAGCCTACCTGCAACTCAGCAGCCTGACATCTGAGGACACTGCCGTCTATTACT GTGCTAGAAGACCTCTCGGACTACCTTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTC A (SEQ ID NO: 165) [00327] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 158 is set forth in SEQ ID NO: 166, which is provided below. GACATCGTGCTGAGCCAGAGCCCTAGCAGCCTGGCCGTGTCTGTGGGCGAGAATGTGACAATGAACT GCAAGAGCTCTCAGAGGCTGCTGGACAGCAGCAACCAGCGGAACTACCTGGCTTGGTACCAGCAAAA GCCCGGCCAGAGCCCCCAGCTGCTGATTTTCTGGGCCTCCACCAGAGAGAGCGGCGTGCCAGATAGA TTCACCGGCAGCGGCAGCGGCACTGATTTCACCCTGACCATCAGAAGCGTGAAAGCTGAAGATCTGG CCGTGTACTACTGCCAGCAGTACTACAGCTACCCCTTTACATTCGGCAGCGGCACCAAGCTGGAAAT CAAAGGAGGCGGCGGATCCGGCGGTGGCGGATCTGGCGGAGGCGGCAGCGAAGTGCAGCTTCAACAA TCTGGCGCCGAGCACGTGAAGCCTGGCACAAGCGTGCGGCTGAGCTGCACCGCCAGCGGATTTAACA TCAAGGACGCCTATATCCACTGGGTCAAGCAGAGACCTGAGCAGGGACTGGAATGGATCGGCGGCAT CGACCCCGAGAACGACAACACCGAGTACGACCCTAAGTTCCCTGGCAAGGCCACCGTTGTGGCCGAT ACATCTTCTAATACCGCCTACCTGCAGCTGTCCTCCCTGACCAGCGAGGACACCGCTGTGTACTATT GTGCCAGACGGCCTCTGGGCCTGCCTTTCGACTACTGGGGCCAGGGCACAACCCTCACAGTGTCCAG C (SEQ ID NO: 166) [00328] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 170; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 171. SEQ ID NO: 170 and SEQ ID NO: 171 are disclosed in Table 10. [00329] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 NAI-1540294631v3 99 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168. SEQ ID NOs: 131, 167, and 168 are provided in Table 10. [00330] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 170. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 170. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 170. [00331] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. SEQ ID NOs: 134, 135, and 169 are provided in Table 10. [00332] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 171. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 171. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 171. [00333] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification NAI-1540294631v3 100 thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. [00334] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 170, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 171. [00335] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00336] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 170, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 171. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 172. SEQ ID NO: 172 is provided in Table 10. [00337] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 NAI-1540294631v3 101 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 170, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 171. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 173. SEQ ID NO: 173 is provided in Table 10. [00338] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 10 (ATA0010) CDRs 1 2 3 VH SYGVH (SEQ ID NO: VVWSGGGTDYNAPFIS NPDSDHYYGYEAMD Q G M A G M G D A G K N
Figure imgf000104_0001
[00339] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 170 is set forth in SEQ ID NO: 174, which is provided below. CAGGTGCAGCTAAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTCTCATTAAATAGCTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGAGTGGCTGGGAGTGGTATGGAGTGGTGGAGGCACAGACTATAATGCACCTTTCATATCCAGACTG AGCATCAGCAAGGACAACTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAAGTTGATGACA NAI-1540294631v3 102 CAGCCATATATTATTGTGCCGGAAATCCAGACTCGGATCATTATTACGGCTACGAGGCTATGGACTA CTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 174) [00340] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 170 is set forth in SEQ ID NO: 175, which is provided below. CAGGTCCAGCTGAAGCAGAGCGGCCCTGGCCTGGTGCAACCTAGCCAGTCCCTGAGCATCACCTGCA CCGTGTCCGGATTTAGCCTGAATAGCTACGGCGTGCATTGGGTGCGGCAAAGCCCCGGCAAAGGCCT GGAATGGCTGGGCGTGGTGTGGAGCGGCGGCGGCACAGACTACAATGCCCCTTTCATCTCTCGGCTG AGCATCAGCAAGGACAACAGTAAGTCTCAGGTTTTCTTCAAGATGAACAGCCTGCAGGTGGACGACA CAGCCATCTACTATTGCGCCGGCAACCCAGACAGCGATCACTACTACGGCTACGAGGCCATGGACTA CTGGGGACAGGGCACCAGCGTGACCGTGTCTAGC (SEQ ID NO: 175) [00341] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 171 is set forth in SEQ ID NO: 176, which is provided below. GACTTCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACAT GTCGAGCAAGTGGAAATATTCACAATTATTTAGCATGGTATCAGCAGAGACAGGGAAAATCTCCTCA GCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGTGGCAGTGGATCA GGACCACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAGGATTTTGGGACTTATTACTGTCAAC ATTTTTGGACTATTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAA (SEQ ID NO: 176) [00342] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 171 is set forth in SEQ ID NO: 177, which is provided below. GACTTCCAGATGACACAGAGCCCTGCCAGCCTCTCCGCCTCTGTGGGCGAGACAGTGACCATTACCT GTAGAGCTTCTGGCAACATCCACAACTACCTGGCCTGGTACCAGCAGAGACAGGGCAAGAGCCCTCA GCTGCTGGTGTACAACGCCAAGACACTGGCTGATGGCGTCCCATCTAGATTCAGCGGCTCCGGCAGC GGCCCCCAGTACAGCCTTAAAATCATCAGCCTGCAACCTGAAGATTTCGGCACCTACTATTGCCAGC ACTTCTGGACCATCCCCTTCACCTTTGGCAGCGGAACCAAGCTGGAGATCAAG (SEQ ID NO: 177) [00343] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 172 is set forth in SEQ ID NO: 178, which is provided below. CAGGTGCAGCTAAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTCTCATTAAATAGCTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGAGTGGCTGGGAGTGGTATGGAGTGGTGGAGGCACAGACTATAATGCACCTTTCATATCCAGACTG AGCATCAGCAAGGACAACTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAAGTTGATGACA CAGCCATATATTATTGTGCCGGAAATCCAGACTCGGATCATTATTACGGCTACGAGGCTATGGACTA NAI-1540294631v3 103 CTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGT GGAGGTGGATCTGACTTCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTG TCACCATCACATGTCGAGCAAGTGGAAATATTCACAATTATTTAGCATGGTATCAGCAGAGACAGGG AAAATCTCCTCAGCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGT GGCAGTGGATCAGGACCACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAGGATTTTGGGACTT ATTACTGTCAACATTTTTGGACTATTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAA (SEQ ID NO: 178) [00344] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 172 is set forth in SEQ ID NO: 179, which is provided below. CAGGTCCAGCTGAAGCAGAGCGGCCCTGGCCTGGTGCAACCTAGCCAGTCCCTGAGCATCACCTGCA CCGTGTCCGGATTTAGCCTGAATAGCTACGGCGTGCATTGGGTGCGGCAAAGCCCCGGCAAAGGCCT GGAATGGCTGGGCGTGGTGTGGAGCGGCGGCGGCACAGACTACAATGCCCCTTTCATCTCTCGGCTG AGCATCAGCAAGGACAACAGTAAGTCTCAGGTTTTCTTCAAGATGAACAGCCTGCAGGTGGACGACA CAGCCATCTACTATTGCGCCGGCAACCCAGACAGCGATCACTACTACGGCTACGAGGCCATGGACTA CTGGGGACAGGGCACCAGCGTGACCGTGTCTAGCGGCGGAGGCGGCTCTGGAGGAGGAGGGTCCGGA GGCGGCGGCAGCGACTTCCAGATGACACAGAGCCCTGCCAGCCTCTCCGCCTCTGTGGGCGAGACAG TGACCATTACCTGTAGAGCTTCTGGCAACATCCACAACTACCTGGCCTGGTACCAGCAGAGACAGGG CAAGAGCCCTCAGCTGCTGGTGTACAACGCCAAGACACTGGCTGATGGCGTCCCATCTAGATTCAGC GGCTCCGGCAGCGGCCCCCAGTACAGCCTTAAAATCATCAGCCTGCAACCTGAAGATTTCGGCACCT ACTATTGCCAGCACTTCTGGACCATCCCCTTCACCTTTGGCAGCGGAACCAAGCTGGAGATCAAG (SEQ ID NO: 179) [00345] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 173 is set forth in SEQ ID NO: 180, which is provided below. GACTTCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACAT GTCGAGCAAGTGGAAATATTCACAATTATTTAGCATGGTATCAGCAGAGACAGGGAAAATCTCCTCA GCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGTGGCAGTGGATCA GGACCACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAGGATTTTGGGACTTATTACTGTCAAC ATTTTTGGACTATTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAAGGTGGAGGTGGATC AGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTGCAGCTAAAGCAGTCAGGACCTGGCCTAGTG CAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAAATAGCTATGGTGTAC ACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGGTATGGAGTGGTGGAGGCAC AGACTATAATGCACCTTTCATATCCAGACTGAGCATCAGCAAGGACAACTCCAAGAGCCAAGTTTTC TTTAAAATGAACAGTCTGCAAGTTGATGACACAGCCATATATTATTGTGCCGGAAATCCAGACTCGG NAI-1540294631v3 104 ATCATTATTACGGCTACGAGGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 180) [00346] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 173 is set forth in SEQ ID NO: 181, which is provided below. GACTTCCAGATGACACAGAGCCCTGCCAGCCTCTCCGCCTCTGTGGGCGAGACAGTGACCATTACCT GTAGAGCTTCTGGCAACATCCACAACTACCTGGCCTGGTACCAGCAGAGACAGGGCAAGAGCCCTCA GCTGCTGGTGTACAACGCCAAGACACTGGCTGATGGCGTCCCATCTAGATTCAGCGGCTCCGGCAGC GGCCCCCAGTACAGCCTTAAAATCATCAGCCTGCAACCTGAAGATTTCGGCACCTACTATTGCCAGC ACTTCTGGACCATCCCCTTCACCTTTGGCAGCGGAACCAAGCTGGAGATCAAGGGCGGAGGCGGCTC TGGAGGAGGAGGGTCCGGAGGCGGCGGCAGCCAGGTCCAGCTGAAGCAGAGCGGCCCTGGCCTGGTG CAACCTAGCCAGTCCCTGAGCATCACCTGCACCGTGTCCGGATTTAGCCTGAATAGCTACGGCGTGC ATTGGGTGCGGCAAAGCCCCGGCAAAGGCCTGGAATGGCTGGGCGTGGTGTGGAGCGGCGGCGGCAC AGACTACAATGCCCCTTTCATCTCTCGGCTGAGCATCAGCAAGGACAACAGTAAGTCTCAGGTTTTC TTCAAGATGAACAGCCTGCAGGTGGACGACACAGCCATCTACTATTGCGCCGGCAACCCAGACAGCG ATCACTACTACGGCTACGAGGCCATGGACTACTGGGGACAGGGCACCAGCGTGACCGTGTCTAGC (SEQ ID NO: 181) [00347] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 183; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 184. SEQ ID NO: 183 and SEQ ID NO: 184 are disclosed in Table 11. [00348] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182. SEQ ID NOs: 41, 24, and 182 are provided in Table 11. [00349] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 183. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about NAI-1540294631v3 105 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 183. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 183. [00350] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. SEQ ID NOs: 26, 27, and 46 are provided in Table 11. [00351] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 184. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 184. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 184. [00352] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. NAI-1540294631v3 106 [00353] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 183, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 184. [00354] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00355] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 184. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 185. SEQ ID NO: 185 is provided in Table 11. [00356] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 184. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 186. SEQ ID NO: 186 is provided in Table 11. [00357] In certain embodiments, the CDRs are identified according to the Kabat numbering system. NAI-1540294631v3 107 Table 11 (ATA011) CDRs 1 2 3 VH DYAIH (SEQ ID NO: VISTYNGNTNYNRKFKD DDY (SEQ ID Q N V L E N V L C L E K I
Figure imgf000110_0001
[00358] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 183 is set forth in SEQ ID NO: 187, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGACTATGCTATACACTGGGTGAAGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACTACAACCGGAAGTTTAAGGACAAG GCCACAATGACTGTTGACAAATCCTCCAGCACAGTTTATATGGAACTTGCCAGATTGACATCTGAGG AATCTGCCATCTATTACTGTGTAAAGGACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTC A (SEQ ID NO: 187) [00359] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 183 is set forth in SEQ ID NO: 188, which is provided below. CAGGTGCAGCTGCAGCAGAGCGGTCCTGAGGTGGTGCGGCCTGGCGTGTCCGTGAAGATCTCTTGCA AGGGCAGCGGATATACCTTTACCGACTACGCCATCCACTGGGTGAAGCAGAGCCACGCCAAGTCCCT CGAGTGGATCGGCGTTATCAGCACATACAACGGCAACACCAACTACAATAGAAAGTTCAAGGACAAA GCCACAATGACCGTGGACAAGAGCAGCAGCACCGTGTACATGGAGCTGGCCAGACTGACCAGCGAGG AAAGCGCTATCTACTACTGCGTGAAAGACGACTACTGGGGCCAGGGCACCAGCGTCACTGTGTCCAG C (SEQ ID NO: 188) NAI-1540294631v3 108 [00360] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 184 is set forth in SEQ ID NO: 189, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCTTCTCTT GCAAGTCAAGTCAGAGCCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGTTGTTACAGAGGCC AGGCCAGTCGCCAAAGCGCCTAATCTATTTGGTGTCTAAACTGGACTCTAGAGTCCCTGACAGGTTC ACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGCACACATTTTCCTCATACGTTCGGAGGAGGGACCAAGTTGGAAATAAA A (SEQ ID NO: 189) [00361] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 184 is set forth in SEQ ID NO: 190, which is provided below. GACGTGGTCATGACCCAGACCCCTCTGACACTGAGCGTGACCATCGGACAACCTGCTTCTTTTAGCT GTAAAAGCAGCCAGTCTCTGCTGGATTCTGATGGCAAGACCTACCTGAACTGGCTGCTGCAAAGACC TGGACAGTCCCCAAAGCGGCTGATCTACCTGGTGTCTAAGCTGGACAGCCGGGTGCCCGATAGATTC ACCGGCAGCGGCTCTGGCACAGATTTCACACTGAAAATTTCTAGAGTGGAAGCCGAGGACCTGGGCG TGTACTATTGCTGGCAGGGCACACATTTCCCCCACACCTTCGGCGGAGGAACAAAGCTGGAAATCAA G (SEQ ID NO: 190) [00362] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 185 is set forth in SEQ ID NO: 191, which is provided below. CAGGTCCAGCTGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCA AGGGTTCCGGCTACACATTCACTGACTATGCTATACACTGGGTGAAGCAGAGTCATGCAAAGAGTCT AGAGTGGATTGGAGTTATTAGTACTTACAATGGTAATACAAACTACAACCGGAAGTTTAAGGACAAG GCCACAATGACTGTTGACAAATCCTCCAGCACAGTTTATATGGAACTTGCCAGATTGACATCTGAGG AATCTGCCATCTATTACTGTGTAAAGGACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTC AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACCCAGACT CCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCTTCTCTTGCAAGTCAAGTCAGAGCCTCT TAGATAGTGATGGAAAGACATATTTGAATTGGTTGTTACAGAGGCCAGGCCAGTCGCCAAAGCGCCT AATCTATTTGGTGTCTAAACTGGACTCTAGAGTCCCTGACAGGTTCACTGGCAGTGGATCAGGGACA GATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGCAAGGCA CACATTTTCCTCATACGTTCGGAGGAGGGACCAAGTTGGAAATAAAA (SEQ ID NO: 191) [00363] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 185 is set forth in SEQ ID NO: 192, which is provided below. CAGGTGCAGCTGCAGCAGAGCGGTCCTGAGGTGGTGCGGCCTGGCGTGTCCGTGAAGATCTCTTGCA AGGGCAGCGGATATACCTTTACCGACTACGCCATCCACTGGGTGAAGCAGAGCCACGCCAAGTCCCT NAI-1540294631v3 109 CGAGTGGATCGGCGTTATCAGCACATACAACGGCAACACCAACTACAATAGAAAGTTCAAGGACAAA GCCACAATGACCGTGGACAAGAGCAGCAGCACCGTGTACATGGAGCTGGCCAGACTGACCAGCGAGG AAAGCGCTATCTACTACTGCGTGAAAGACGACTACTGGGGCCAGGGCACCAGCGTCACTGTGTCCAG CGGCGGCGGCGGCAGCGGCGGAGGCGGCAGCGGCGGCGGAGGCAGCGACGTGGTCATGACCCAGACC CCTCTGACACTGAGCGTGACCATCGGACAACCTGCTTCTTTTAGCTGTAAAAGCAGCCAGTCTCTGC TGGATTCTGATGGCAAGACCTACCTGAACTGGCTGCTGCAAAGACCTGGACAGTCCCCAAAGCGGCT GATCTACCTGGTGTCTAAGCTGGACAGCCGGGTGCCCGATAGATTCACCGGCAGCGGCTCTGGCACA GATTTCACACTGAAAATTTCTAGAGTGGAAGCCGAGGACCTGGGCGTGTACTATTGCTGGCAGGGCA CACATTTCCCCCACACCTTCGGCGGAGGAACAAAGCTGGAAATCAAG (SEQ ID NO: 192) [00364] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 186 is set forth in SEQ ID NO: 193, which is provided below. GATGTTGTGATGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCTTCTCTT GCAAGTCAAGTCAGAGCCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGTTGTTACAGAGGCC AGGCCAGTCGCCAAAGCGCCTAATCTATTTGGTGTCTAAACTGGACTCTAGAGTCCCTGACAGGTTC ACTGGCAGTGGATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAG TTTATTATTGCTGGCAAGGCACACATTTTCCTCATACGTTCGGAGGAGGGACCAAGTTGGAAATAAA AGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAGCTGCAGCAGTCT GGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACACATTCA CTGACTATGCTATACACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATTAG TACTTACAATGGTAATACAAACTACAACCGGAAGTTTAAGGACAAGGCCACAATGACTGTTGACAAA TCCTCCAGCACAGTTTATATGGAACTTGCCAGATTGACATCTGAGGAATCTGCCATCTATTACTGTG TAAAGGACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 193) [00365] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 186 is set forth in SEQ ID NO: 194, which is provided below. GACGTGGTCATGACCCAGACCCCTCTGACACTGAGCGTGACCATCGGACAACCTGCTTCTTTTAGCT GTAAAAGCAGCCAGTCTCTGCTGGATTCTGATGGCAAGACCTACCTGAACTGGCTGCTGCAAAGACC TGGACAGTCCCCAAAGCGGCTGATCTACCTGGTGTCTAAGCTGGACAGCCGGGTGCCCGATAGATTC ACCGGCAGCGGCTCTGGCACAGATTTCACACTGAAAATTTCTAGAGTGGAAGCCGAGGACCTGGGCG TGTACTATTGCTGGCAGGGCACACATTTCCCCCACACCTTCGGCGGAGGAACAAAGCTGGAAATCAA GGGCGGCGGCGGCAGCGGCGGAGGCGGCAGCGGCGGCGGAGGCAGCCAGGTGCAGCTGCAGCAGAGC GGTCCTGAGGTGGTGCGGCCTGGCGTGTCCGTGAAGATCTCTTGCAAGGGCAGCGGATATACCTTTA CCGACTACGCCATCCACTGGGTGAAGCAGAGCCACGCCAAGTCCCTCGAGTGGATCGGCGTTATCAG CACATACAACGGCAACACCAACTACAATAGAAAGTTCAAGGACAAAGCCACAATGACCGTGGACAAG NAI-1540294631v3 110 AGCAGCAGCACCGTGTACATGGAGCTGGCCAGACTGACCAGCGAGGAAAGCGCTATCTACTACTGCG TGAAAGACGACTACTGGGGCCAGGGCACCAGCGTCACTGTGTCCAGC (SEQ ID NO: 194) [00366] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 201; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 202. SEQ ID NO: 201 and SEQ ID NO: 202 are disclosed in Table 12. [00367] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197. SEQ ID NOs: 195-197 are provided in Table 12. [00368] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 201. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 201. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 201. [00369] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200. SEQ ID NOs: 198-200 are provided in Table 12. [00370] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 202. For example, the VL comprises an amino NAI-1540294631v3 111 acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 202. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 202. [00371] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200. [00372] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 201, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 202. [00373] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00374] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200. In certain NAI-1540294631v3 112 embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 202. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 203. SEQ ID NO: 203 is provided in Table 12. [00375] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 202. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 204. SEQ ID NO: 204 is provided in Table 12. [00376] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 12 (ATA012) CDRs 1 2 3 V SYYMS (SEQ ID NO: AINFYGGLAYYPDTVKG HYRYDNAMDY Q G G Y G G T Y Y G
Figure imgf000115_0001
113 Amino GSGGGGSGGGGSDVKLVESGGGLVKLGGSLKLSCAASGFTFSSYYMSWVRQTPEK Acid RLDLVAAINFYGGLAYYPDTVKGRFTISRDNAKNTLYLQMNSLKSEDTALYYCVR HYRYDNAMDYWGQGTSVTVSS (SEQ ID NO 204)
Figure imgf000116_0001
sequence of SEQ ID NO: 201 is set forth in SEQ ID NO: 205, which is provided below. GACGTTAAGCTCGTGGAGTCTGGGGGAGGCTTAGTGAAGCTTGGAGGGTCCCTGAAACTCTCCTGTG CAGCCTCTGGATTCACTTTCAGTAGCTATTACATGTCTTGGGTTCGCCAGACTCCAGAGAAGAGGCT GGACTTGGTCGCAGCCATAAATTTTTATGGTGGTCTTGCCTACTATCCAGACACTGTGAAGGGCCGA TTCACCATCTCCAGAGACAATGCCAAGAACACCCTGTACCTGCAAATGAACAGTCTGAAGTCTGAGG ACACAGCCTTGTATTACTGTGTGAGACATTATAGGTACGACAATGCTATGGACTACTGGGGTCAAGG AACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 205) [00378] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 201 is set forth in SEQ ID NO: 206, which is provided below. GACGTGAAACTGGTTGAGAGCGGAGGCGGCCTGGTGAAACTGGGAGGAAGCCTGAAACTGTCTTGTG CCGCCAGCGGCTTCACCTTTAGCAGCTATTACATGTCCTGGGTGCGGCAGACACCAGAGAAGAGACT GGACCTGGTGGCCGCTATCAACTTCTACGGCGGCCTGGCCTACTACCCTGACACCGTGAAGGGCAGA TTCACCATCAGCAGAGACAACGCCAAGAACACCCTGTACCTGCAGATGAACAGCCTCAAGAGCGAGG ACACTGCCCTGTATTACTGCGTGAGGCACTACCGGTACGACAACGCCATGGACTACTGGGGCCAGGG CACCAGCGTGACCGTGTCCTCT (SEQ ID NO: 206) [00379] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 202 is set forth in SEQ ID NO: 207, which is provided below. GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAATTAGAGACAGGGTCAACATCACCT GCAAGGCCAGTCAGAATGTGGGTACTGCTGTAGCCTGGTATCAACAGAGACCAGGACAATCTCCTAA ACTACTGATTTACTCGGCATCCAATCGGTACAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCT GGGACAGATTTCACTCTCACCATCAGCAATATGCAGTCTGAAGACCTGACAAGTTATTTCTGCCAGC AATATAGTAGTTATCCTCTCACGTTCGGTGTTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 207) [00380] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 202 is set forth in SEQ ID NO: 208, which is provided below. GATATCGTGATGACCCAGAGCCAGAAGTTCATGAGCACAAGCATCAGAGATAGAGTGAACATCACCT GCAAGGCTTCCCAGAACGTGGGCACAGCCGTGGCTTGGTACCAGCAAAGACCTGGCCAGTCCCCTAA GCTGCTGATCTACAGCGCCAGCAATAGATACTCTGGGGTCCCCGACCGGTTTACCGGCAGCGGCAGC GGCACAGATTTCACCCTGACCATTTCTAATATGCAGAGCGAAGATCTGACAAGCTACTTCTGCCAGC NAI-1540294631v3 114 AATACTCTTCTTACCCCCTGACATTCGGCGTGGGAACCAAGCTGGAACTGAAG (SEQ ID NO: 208) [00381] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 203 is set forth in SEQ ID NO: 209, which is provided below. GACGTTAAGCTCGTGGAGTCTGGGGGAGGCTTAGTGAAGCTTGGAGGGTCCCTGAAACTCTCCTGTG CAGCCTCTGGATTCACTTTCAGTAGCTATTACATGTCTTGGGTTCGCCAGACTCCAGAGAAGAGGCT GGACTTGGTCGCAGCCATAAATTTTTATGGTGGTCTTGCCTACTATCCAGACACTGTGAAGGGCCGA TTCACCATCTCCAGAGACAATGCCAAGAACACCCTGTACCTGCAAATGAACAGTCTGAAGTCTGAGG ACACAGCCTTGTATTACTGTGTGAGACATTATAGGTACGACAATGCTATGGACTACTGGGGTCAAGG AACCTCAGTCACCGTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCT GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAATTAGAGACAGGGTCAACATCACCT GCAAGGCCAGTCAGAATGTGGGTACTGCTGTAGCCTGGTATCAACAGAGACCAGGACAATCTCCTAA ACTACTGATTTACTCGGCATCCAATCGGTACAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCT GGGACAGATTTCACTCTCACCATCAGCAATATGCAGTCTGAAGACCTGACAAGTTATTTCTGCCAGC AATATAGTAGTTATCCTCTCACGTTCGGTGTTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 209) [00382] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 203 is set forth in SEQ ID NO: 210, which is provided below. GACGTGAAACTGGTTGAGAGCGGAGGCGGCCTGGTGAAACTGGGAGGAAGCCTGAAACTGTCTTGTG CCGCCAGCGGCTTCACCTTTAGCAGCTATTACATGTCCTGGGTGCGGCAGACACCAGAGAAGAGACT GGACCTGGTGGCCGCTATCAACTTCTACGGCGGCCTGGCCTACTACCCTGACACCGTGAAGGGCAGA TTCACCATCAGCAGAGACAACGCCAAGAACACCCTGTACCTGCAGATGAACAGCCTCAAGAGCGAGG ACACTGCCCTGTATTACTGCGTGAGGCACTACCGGTACGACAACGCCATGGACTACTGGGGCCAGGG CACCAGCGTGACCGTGTCCTCTGGCGGAGGCGGCAGCGGCGGCGGCGGAAGCGGCGGCGGAGGCAGC GATATCGTGATGACCCAGAGCCAGAAGTTCATGAGCACAAGCATCAGAGATAGAGTGAACATCACCT GCAAGGCTTCCCAGAACGTGGGCACAGCCGTGGCTTGGTACCAGCAAAGACCTGGCCAGTCCCCTAA GCTGCTGATCTACAGCGCCAGCAATAGATACTCTGGGGTCCCCGACCGGTTTACCGGCAGCGGCAGC GGCACAGATTTCACCCTGACCATTTCTAATATGCAGAGCGAAGATCTGACAAGCTACTTCTGCCAGC AATACTCTTCTTACCCCCTGACATTCGGCGTGGGAACCAAGCTGGAACTGAAG (SEQ ID NO: 210) [00383] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 204 is set forth in SEQ ID NO: 211, which is provided below. NAI-1540294631v3 115 GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAATTAGAGACAGGGTCAACATCACCT GCAAGGCCAGTCAGAATGTGGGTACTGCTGTAGCCTGGTATCAACAGAGACCAGGACAATCTCCTAA ACTACTGATTTACTCGGCATCCAATCGGTACAGTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCT GGGACAGATTTCACTCTCACCATCAGCAATATGCAGTCTGAAGACCTGACAAGTTATTTCTGCCAGC AATATAGTAGTTATCCTCTCACGTTCGGTGTTGGGACCAAGCTGGAGCTGAAAGGTGGAGGTGGATC AGGTGGAGGTGGATCTGGTGGAGGTGGATCTGACGTTAAGCTCGTGGAGTCTGGGGGAGGCTTAGTG AAGCTTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAGCTATTACATGT CTTGGGTTCGCCAGACTCCAGAGAAGAGGCTGGACTTGGTCGCAGCCATAAATTTTTATGGTGGTCT TGCCTACTATCCAGACACTGTGAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACACCCTG TACCTGCAAATGAACAGTCTGAAGTCTGAGGACACAGCCTTGTATTACTGTGTGAGACATTATAGGT ACGACAATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 211) [00384] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 204 is set forth in SEQ ID NO: 212, which is provided below. GATATCGTGATGACCCAGAGCCAGAAGTTCATGAGCACAAGCATCAGAGATAGAGTGAACATCACCT GCAAGGCTTCCCAGAACGTGGGCACAGCCGTGGCTTGGTACCAGCAAAGACCTGGCCAGTCCCCTAA GCTGCTGATCTACAGCGCCAGCAATAGATACTCTGGGGTCCCCGACCGGTTTACCGGCAGCGGCAGC GGCACAGATTTCACCCTGACCATTTCTAATATGCAGAGCGAAGATCTGACAAGCTACTTCTGCCAGC AATACTCTTCTTACCCCCTGACATTCGGCGTGGGAACCAAGCTGGAACTGAAGGGCGGAGGCGGCAG CGGCGGCGGCGGAAGCGGCGGCGGAGGCAGCGACGTGAAACTGGTTGAGAGCGGAGGCGGCCTGGTG AAACTGGGAGGAAGCCTGAAACTGTCTTGTGCCGCCAGCGGCTTCACCTTTAGCAGCTATTACATGT CCTGGGTGCGGCAGACACCAGAGAAGAGACTGGACCTGGTGGCCGCTATCAACTTCTACGGCGGCCT GGCCTACTACCCTGACACCGTGAAGGGCAGATTCACCATCAGCAGAGACAACGCCAAGAACACCCTG TACCTGCAGATGAACAGCCTCAAGAGCGAGGACACTGCCCTGTATTACTGCGTGAGGCACTACCGGT ACGACAACGCCATGGACTACTGGGGCCAGGGCACCAGCGTGACCGTGTCCTCT (SEQ ID NO: 212) [00385] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 215; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 216. SEQ ID NO: 215 and SEQ ID NO: 216 are disclosed in Table 13. [00386] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 NAI-1540294631v3 116 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168. SEQ ID NOs: 213, 167, and 168 are provided in Table 13. [00387] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 215. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 215. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 215. [00388] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. SEQ ID NOs: 214, 135, and 169 are provided in Table 13. [00389] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 216. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 216. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 216. [00390] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification NAI-1540294631v3 117 thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. [00391] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 215, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 216. [00392] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00393] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 215, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 216. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 217. SEQ ID NO: 217 is provided in Table 13. [00394] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 NAI-1540294631v3 118 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 215, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 216. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 218. SEQ ID NO: 218 is provided in Table 13. [00395] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 13 (ATA013) CDRs 1 2 3 VH NYGIN (SEQ ID NO: VVWSGGGTDYNAPFIS NPDSDHYYGYEAMD Q G M A G M G D A G K N
Figure imgf000121_0001
[00396] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 215 is set forth in SEQ ID NO: 219, which is provided below. CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTTTCATTAAATAACTATGGTATAAACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGAGTGGCTGGGAGTGGTATGGAGTGGTGGAGGCACAGACTATAATGCACCTTTCATATCCAGACTG AGCATCAGCAAGGACAACTCCAAGAGCCAAGTTTTCTTTAAAATGAGCAGTCTGCAAGTTGATGACA NAI-1540294631v3 119 CAGCCATATATTATTGTGCCGGAAATCCAGACTCGGATCATTACTACGGCTACGAGGCTATGGACTA CTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 219) [00397] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 215 is set forth in SEQ ID NO: 220, which is provided below. CAAGTGCAGCTGAAGCAGAGCGGACCTGGCCTGGTGCAGCCTAGCCAGTCCCTGAGCATCACCTGTA CCGTGAGCGGATTTTCTCTGAACAACTACGGCATCAACTGGGTGCGGCAGAGCCCCGGCAAGGGCCT GGAATGGCTGGGCGTGGTGTGGTCCGGCGGCGGCACTGATTATAATGCCCCTTTCATCTCCAGACTG TCCATTTCTAAGGACAACAGCAAGAGCCAGGTGTTCTTCAAGATGAGCAGCCTGCAGGTGGACGACA CCGCCATCTACTATTGCGCCGGCAACCCCGACAGCGACCACTACTACGGCTACGAGGCCATGGACTA CTGGGGCCAGGGCACAAGCGTGACCGTGTCCAGC (SEQ ID NO: 220) [00398] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 216 is set forth in SEQ ID NO: 221, which is provided below. GACTTCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCTTCACTT GTCGAGCAAGTGGAGATATTCACAATTTTTTAGCATGGTATCAGCAGAGACAGGGAAAGTCTCCTCA GCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGTGGCAGTGCATCA GGAACACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAGGATTTTGGGACTTATTACTGTCAAC ATTTTTGGACTATTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAA (SEQ ID NO: 221) [00399] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 216 is set forth in SEQ ID NO: 222, which is provided below. GATTTCCAGATGACCCAGTCTCCAGCCAGCCTGTCTGCTTCTGTGGGCGAGACAGTCACCTTTACAT GCAGAGCCAGCGGCGACATCCACAACTTCCTGGCCTGGTACCAGCAGCGGCAGGGAAAAAGCCCTCA GCTGCTGGTTTACAATGCCAAGACCCTGGCTGATGGAGTGCCTAGCAGATTCAGCGGCAGCGCCTCT GGCACACAGTACAGCCTGAAAATCATCAGCCTCCAACCTGAGGATTTCGGCACCTACTACTGCCAAC ACTTCTGGACCATCCCCTTCACATTTGGCAGCGGCACCAAGCTGGAAATCAAG (SEQ ID NO: 222) [00400] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 217 is set forth in SEQ ID NO: 223, which is provided below. CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTTTCATTAAATAACTATGGTATAAACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGAGTGGCTGGGAGTGGTATGGAGTGGTGGAGGCACAGACTATAATGCACCTTTCATATCCAGACTG AGCATCAGCAAGGACAACTCCAAGAGCCAAGTTTTCTTTAAAATGAGCAGTCTGCAAGTTGATGACA CAGCCATATATTATTGTGCCGGAAATCCAGACTCGGATCATTACTACGGCTACGAGGCTATGGACTA NAI-1540294631v3 120 CTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGT GGAGGTGGATCTGACTTCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTG TCACCTTCACTTGTCGAGCAAGTGGAGATATTCACAATTTTTTAGCATGGTATCAGCAGAGACAGGG AAAGTCTCCTCAGCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGT GGCAGTGCATCAGGAACACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAGGATTTTGGGACTT ATTACTGTCAACATTTTTGGACTATTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAA (SEQ ID NO: 223) [00401] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 217 is set forth in SEQ ID NO: 224, which is provided below. CAAGTGCAGCTGAAGCAGAGCGGACCTGGCCTGGTGCAGCCTAGCCAGTCCCTGAGCATCACCTGTA CCGTGAGCGGATTTTCTCTGAACAACTACGGCATCAACTGGGTGCGGCAGAGCCCCGGCAAGGGCCT GGAATGGCTGGGCGTGGTGTGGTCCGGCGGCGGCACTGATTATAATGCCCCTTTCATCTCCAGACTG TCCATTTCTAAGGACAACAGCAAGAGCCAGGTGTTCTTCAAGATGAGCAGCCTGCAGGTGGACGACA CCGCCATCTACTATTGCGCCGGCAACCCCGACAGCGACCACTACTACGGCTACGAGGCCATGGACTA CTGGGGCCAGGGCACAAGCGTGACCGTGTCCAGCGGCGGCGGAGGCTCTGGCGGTGGCGGAAGCGGC GGCGGCGGAAGCGATTTCCAGATGACCCAGTCTCCAGCCAGCCTGTCTGCTTCTGTGGGCGAGACAG TCACCTTTACATGCAGAGCCAGCGGCGACATCCACAACTTCCTGGCCTGGTACCAGCAGCGGCAGGG AAAAAGCCCTCAGCTGCTGGTTTACAATGCCAAGACCCTGGCTGATGGAGTGCCTAGCAGATTCAGC GGCAGCGCCTCTGGCACACAGTACAGCCTGAAAATCATCAGCCTCCAACCTGAGGATTTCGGCACCT ACTACTGCCAACACTTCTGGACCATCCCCTTCACATTTGGCAGCGGCACCAAGCTGGAAATCAAG (SEQ ID NO: 224) [00402] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 218 is set forth in SEQ ID NO: 225, which is provided below. GACTTCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCTTCACTT GTCGAGCAAGTGGAGATATTCACAATTTTTTAGCATGGTATCAGCAGAGACAGGGAAAGTCTCCTCA GCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGTGGCAGTGCATCA GGAACACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAGGATTTTGGGACTTATTACTGTCAAC ATTTTTGGACTATTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAAGGTGGAGGTGGATC AGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTG CAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTTTCATTAAATAACTATGGTATAA ACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGGTATGGAGTGGTGGAGGCAC AGACTATAATGCACCTTTCATATCCAGACTGAGCATCAGCAAGGACAACTCCAAGAGCCAAGTTTTC TTTAAAATGAGCAGTCTGCAAGTTGATGACACAGCCATATATTATTGTGCCGGAAATCCAGACTCGG NAI-1540294631v3 121 ATCATTACTACGGCTACGAGGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 225) [00403] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 218 is set forth in SEQ ID NO: 226, which is provided below. GATTTCCAGATGACCCAGTCTCCAGCCAGCCTGTCTGCTTCTGTGGGCGAGACAGTCACCTTTACAT GCAGAGCCAGCGGCGACATCCACAACTTCCTGGCCTGGTACCAGCAGCGGCAGGGAAAAAGCCCTCA GCTGCTGGTTTACAATGCCAAGACCCTGGCTGATGGAGTGCCTAGCAGATTCAGCGGCAGCGCCTCT GGCACACAGTACAGCCTGAAAATCATCAGCCTCCAACCTGAGGATTTCGGCACCTACTACTGCCAAC ACTTCTGGACCATCCCCTTCACATTTGGCAGCGGCACCAAGCTGGAAATCAAGGGCGGCGGAGGCTC TGGCGGTGGCGGAAGCGGCGGCGGCGGAAGCCAAGTGCAGCTGAAGCAGAGCGGACCTGGCCTGGTG CAGCCTAGCCAGTCCCTGAGCATCACCTGTACCGTGAGCGGATTTTCTCTGAACAACTACGGCATCA ACTGGGTGCGGCAGAGCCCCGGCAAGGGCCTGGAATGGCTGGGCGTGGTGTGGTCCGGCGGCGGCAC TGATTATAATGCCCCTTTCATCTCCAGACTGTCCATTTCTAAGGACAACAGCAAGAGCCAGGTGTTC TTCAAGATGAGCAGCCTGCAGGTGGACGACACCGCCATCTACTATTGCGCCGGCAACCCCGACAGCG ACCACTACTACGGCTACGAGGCCATGGACTACTGGGGCCAGGGCACAAGCGTGACCGTGTCCAGC (SEQ ID NO: 226) [00404] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 233; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 234. SEQ ID NO: 233 and SEQ ID NO: 234 are disclosed in Table 14. [00405] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229. SEQ ID NOs: 227-229 are provided in Table 14. [00406] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 233. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about NAI-1540294631v3 122 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 233. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 233. [00407] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232. SEQ ID NOs: 230-232 are provided in Table 14. [00408] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 234. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 234. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 234. [00409] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232. NAI-1540294631v3 123 [00410] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 233, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 234. [00411] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00412] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 233, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 234. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 235. SEQ ID NO: 235 is provided in Table 14. [00413] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 233, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 234. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 236. SEQ ID NO: 236 is provided in Table 14. [00414] In certain embodiments, the CDRs are identified according to the Kabat numbering system. NAI-1540294631v3 124 Table 14 (ATA014) CDRs 1 2 3 VH NYWMH (SEQ ID NO: NIYPGSCTSNYDERFKN GNYDYYFDY (SEQ Q S Q H S Q W F H G Q R
Figure imgf000127_0001
[00415] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 233 is set forth in SEQ ID NO: 237, which is provided below. CAGGTCCAACTGCAGCAACCTGGGTCTGAGCTGGTGAGGCCTGGAGTTTCAGTGAACCTGTCCTGCA AGGCTTCTGGCTACACTTTCACCAACTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCT TGAATGGATTGGAAATATTTATCCTGGTTCTTGTACTTCTAACTACGATGAGAGGTTCAAGAACAAG GTCACACTGACTGTAGACACATCTTCCAGTACAGCCTACATGCAGTTCAGCAGCCTGACATCTGAGG ACTCTGCGGTCTATTACTGTTCAAGAGGAAACTATGATTACTATTTTGACTACTGGGGCCAAGGCAC CACTCTCACAGTCTCCTCA (SEQ ID NO: 237) [00416] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 233 is set forth in SEQ ID NO: 238, which is provided below. CAAGTGCAGCTGCAACAGCCTGGCAGCGAGCTGGTGCGGCCTGGAGTGTCCGTGAACCTGTCTTGTA AAGCTTCTGGCTACACCTTCACCAACTACTGGATGCACTGGGTCAAGCAGAGACCAGGCCAGGGTCT CGAGTGGATCGGCAACATCTACCCTGGCAGCTGCACCAGCAACTACGACGAGAGATTTAAAAACAAG GTGACCCTGACCGTGGACACCTCCTCTAGCACCGCCTACATGCAGTTCAGCTCCCTGACATCTGAAG ATAGCGCCGTGTACTACTGCAGCAGAGGCAATTATGACTACTACTTCGACTACTGGGGCCAGGGCAC CACACTGACCGTGTCCAGC (SEQ ID NO: 238) NAI-1540294631v3 125 [00417] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 234 is set forth in SEQ ID NO: 239, which is provided below. GACATCCAGATGAACCAGTCTCCATCCAGTCTGTCTGCATCCCTTGGAGACACAGTTACCATCACTT GCCATGCCAGTCAGAACATTAATGTTTGGTTAAGCTGGTACCAGCAGAGACCAGGAAATATTCCTAA CCTATTGATCTATAAGGCTTCCAACTTACACACGGGCGTCCCGTCAAGGTTTAGTGGCAGTGGATCT GGAACAGGTTTCACGTTAACCATCAGCAGCCTGCAGCCTGAAGACATTGCCACTTACTTCTGTCAAC AGGGTCAAACTTATCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 239) [00418] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 234 is set forth in SEQ ID NO: 240, which is provided below. GACATCCAGATGAACCAGAGCCCTAGCAGCCTGAGCGCTAGCCTGGGAGATACCGTGACAATCACCT GCCACGCCAGCCAGAACATCAACGTGTGGCTGAGCTGGTACCAGCAAAGACCCGGCAATATTCCTAA CCTGCTGATCTACAAGGCCTCTAATCTGCACACCGGCGTTCCTTCTCGGTTTAGCGGATCTGGATCC GGCACAGGCTTCACACTGACAATCAGCAGCCTGCAGCCCGAGGATATCGCCACATATTTCTGCCAGC AGGGACAGACCTACCCCTGGACATTCGGCGGCGGCACCAAGCTGGAAATCAAG (SEQ ID NO: 240) [00419] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 235 is set forth in SEQ ID NO: 241, which is provided below. CAGGTCCAACTGCAGCAACCTGGGTCTGAGCTGGTGAGGCCTGGAGTTTCAGTGAACCTGTCCTGCA AGGCTTCTGGCTACACTTTCACCAACTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCT TGAATGGATTGGAAATATTTATCCTGGTTCTTGTACTTCTAACTACGATGAGAGGTTCAAGAACAAG GTCACACTGACTGTAGACACATCTTCCAGTACAGCCTACATGCAGTTCAGCAGCCTGACATCTGAGG ACTCTGCGGTCTATTACTGTTCAAGAGGAAACTATGATTACTATTTTGACTACTGGGGCCAAGGCAC CACTCTCACAGTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGAC ATCCAGATGAACCAGTCTCCATCCAGTCTGTCTGCATCCCTTGGAGACACAGTTACCATCACTTGCC ATGCCAGTCAGAACATTAATGTTTGGTTAAGCTGGTACCAGCAGAGACCAGGAAATATTCCTAACCT ATTGATCTATAAGGCTTCCAACTTACACACGGGCGTCCCGTCAAGGTTTAGTGGCAGTGGATCTGGA ACAGGTTTCACGTTAACCATCAGCAGCCTGCAGCCTGAAGACATTGCCACTTACTTCTGTCAACAGG GTCAAACTTATCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 241) [00420] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 235 is set forth in SEQ ID NO: 242, which is provided below. CAAGTGCAGCTGCAACAGCCTGGCAGCGAGCTGGTGCGGCCTGGAGTGTCCGTGAACCTGTCTTGTA AAGCTTCTGGCTACACCTTCACCAACTACTGGATGCACTGGGTCAAGCAGAGACCAGGCCAGGGTCT NAI-1540294631v3 126 CGAGTGGATCGGCAACATCTACCCTGGCAGCTGCACCAGCAACTACGACGAGAGATTTAAAAACAAG GTGACCCTGACCGTGGACACCTCCTCTAGCACCGCCTACATGCAGTTCAGCTCCCTGACATCTGAAG ATAGCGCCGTGTACTACTGCAGCAGAGGCAATTATGACTACTACTTCGACTACTGGGGCCAGGGCAC CACACTGACCGTGTCCAGCGGAGGCGGCGGCAGCGGCGGCGGCGGCAGTGGCGGAGGCGGCAGCGAC ATCCAGATGAACCAGAGCCCTAGCAGCCTGAGCGCTAGCCTGGGAGATACCGTGACAATCACCTGCC ACGCCAGCCAGAACATCAACGTGTGGCTGAGCTGGTACCAGCAAAGACCCGGCAATATTCCTAACCT GCTGATCTACAAGGCCTCTAATCTGCACACCGGCGTTCCTTCTCGGTTTAGCGGATCTGGATCCGGC ACAGGCTTCACACTGACAATCAGCAGCCTGCAGCCCGAGGATATCGCCACATATTTCTGCCAGCAGG GACAGACCTACCCCTGGACATTCGGCGGCGGCACCAAGCTGGAAATCAAG (SEQ ID NO: 242) [00421] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 236 is set forth in SEQ ID NO: 243, which is provided below. GACATCCAGATGAACCAGTCTCCATCCAGTCTGTCTGCATCCCTTGGAGACACAGTTACCATCACTT GCCATGCCAGTCAGAACATTAATGTTTGGTTAAGCTGGTACCAGCAGAGACCAGGAAATATTCCTAA CCTATTGATCTATAAGGCTTCCAACTTACACACGGGCGTCCCGTCAAGGTTTAGTGGCAGTGGATCT GGAACAGGTTTCACGTTAACCATCAGCAGCCTGCAGCCTGAAGACATTGCCACTTACTTCTGTCAAC AGGGTCAAACTTATCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAAGGTGGAGGTGGATC AGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAACTGCAGCAACCTGGGTCTGAGCTGGTG AGGCCTGGAGTTTCAGTGAACCTGTCCTGCAAGGCTTCTGGCTACACTTTCACCAACTACTGGATGC ACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAATGGATTGGAAATATTTATCCTGGTTCTTGTAC TTCTAACTACGATGAGAGGTTCAAGAACAAGGTCACACTGACTGTAGACACATCTTCCAGTACAGCC TACATGCAGTTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTTCAAGAGGAAACTATG ATTACTATTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA (SEQ ID NO: 243) [00422] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 236 is set forth in SEQ ID NO: 244, which is provided below. GACATCCAGATGAACCAGAGCCCTAGCAGCCTGAGCGCTAGCCTGGGAGATACCGTGACAATCACCT GCCACGCCAGCCAGAACATCAACGTGTGGCTGAGCTGGTACCAGCAAAGACCCGGCAATATTCCTAA CCTGCTGATCTACAAGGCCTCTAATCTGCACACCGGCGTTCCTTCTCGGTTTAGCGGATCTGGATCC GGCACAGGCTTCACACTGACAATCAGCAGCCTGCAGCCCGAGGATATCGCCACATATTTCTGCCAGC AGGGACAGACCTACCCCTGGACATTCGGCGGCGGCACCAAGCTGGAAATCAAGGGAGGCGGCGGCAG CGGCGGCGGCGGCAGTGGCGGAGGCGGCAGCCAAGTGCAGCTGCAACAGCCTGGCAGCGAGCTGGTG CGGCCTGGAGTGTCCGTGAACCTGTCTTGTAAAGCTTCTGGCTACACCTTCACCAACTACTGGATGC ACTGGGTCAAGCAGAGACCAGGCCAGGGTCTCGAGTGGATCGGCAACATCTACCCTGGCAGCTGCAC CAGCAACTACGACGAGAGATTTAAAAACAAGGTGACCCTGACCGTGGACACCTCCTCTAGCACCGCC NAI-1540294631v3 127 TACATGCAGTTCAGCTCCCTGACATCTGAAGATAGCGCCGTGTACTACTGCAGCAGAGGCAATTATG ACTACTACTTCGACTACTGGGGCCAGGGCACCACACTGACCGTGTCCAGC (SEQ ID NO: 244) [00423] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 246; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 247. SEQ ID NO: 246 and SEQ ID NO: 247 are disclosed in Table 15. [00424] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115. SEQ ID NOs: 113, 245, and 115 are provided in Table 15. [00425] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 247. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 247. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 247. [00426] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. SEQ ID NOs: 116-118 are provided in Table 15. [00427] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 247. For example, the VL comprises an amino NAI-1540294631v3 128 acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 247. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 247. [00428] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. [00429] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 246, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 247. [00430] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00431] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. In certain NAI-1540294631v3 129 embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 247. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 248. SEQ ID NO: 248 is provided in Table 15. [00432] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 247. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 249. SEQ ID NO: 249 is provided in Table 15. [00433] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 15 (ATA015) CDRs 1 2 3 V TYGVH (SEQ ID NO: VIWSGGSTDYNAAFIS RGNDGPPFAY Q G Q D G Q Y Y D G
Figure imgf000132_0001
130 Amino GSGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTTYGVHWVRQSPGK Acid GLDWLGVIWSGGSTDYNAAFISRLSISKDNSKSQVFFKMNSLQTNDTAIYYCARR GNDGPPFAYWGQGTLVTVSA (SEQ ID NO 249)
Figure imgf000133_0001
sequence of SEQ ID NO: 246 is set forth in SEQ ID NO: 250, which is provided below. CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTCTCATTAACTACCTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGATTGGCTGGGAGTGATATGGAGTGGTGGAAGCACAGACTATAATGCAGCTTTCATATCCAGACTG AGCATCAGCAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAAACTAATGACA CAGCCATATATTACTGTGCCAGAAGAGGGAATGATGGTCCTCCGTTTGCTTACTGGGGCCAAGGGAC TCTGGTCACTGTCTCTGCA (SEQ ID NO: 250) [00435] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 246 is set forth in SEQ ID NO: 251, which is provided below. CAAGTGCAGCTGAAGCAGAGCGGCCCTGGACTGGTGCAGCCTTCTCAGAGCCTCTCCATCACCTGCA CCGTGTCCGGCTTCAGCCTGACAACATACGGCGTGCACTGGGTGCGGCAGAGCCCCGGCAAGGGCCT GGACTGGCTGGGCGTGATCTGGTCCGGCGGCTCCACAGATTACAACGCCGCTTTTATCAGCAGACTG TCCATCAGCAAGGACAACAGCAAGAGCCAGGTGTTCTTCAAGATGAACAGCCTGCAGACCAATGACA CCGCCATCTACTACTGCGCCAGAAGGGGCAACGACGGCCCACCTTTCGCCTACTGGGGCCAGGGAAC ACTGGTGACCGTTAGTGCT (SEQ ID NO: 251) [00436] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 247 is set forth in SEQ ID NO: 252, which is provided below. GACATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTT GTCGGGCAAGTCAGGAAATTAGTGGTTACTTAAGCTGGCTTCAGAAGAAACCAGATGGAACTATTAA ACGCCTGATCTACGCCGCATCCACTTTAGATTCTGGTGTCCCAAAAAGGTTCAGTGGCAGTAGGTCT GGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGCAGACTATTACTGTCTAC AATATGTTAGTTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 252) [00437] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 247 is set forth in SEQ ID NO: 253, which is provided below. GATATCCAGATGACCCAGTCTCCTAGCAGCCTGAGCGCTAGCCTGGGCGAGAGAGTGTCCCTGACAT GTAGAGCCAGCCAGGAGATCAGCGGCTACCTGAGCTGGCTGCAGAAAAAGCCCGACGGCACCATCAA GAGACTGATCTACGCCGCCTCTACCCTGGATAGCGGCGTCCCCAAGCGGTTCAGCGGAAGCCGGAGC GGTTCTGATTACAGCCTGACCATTTCTAGCCTTGAAAGCGAGGACTTCGCCGACTACTATTGCCTGC NAI-1540294631v3 131 AATACGTGTCTTATCCTCTGACTTTTGGAGCCGGCACCAAGCTGGAACTGAAA (SEQ ID NO: 253) [00438] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 248 is set forth in SEQ ID NO: 254, which is provided below. CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCA CAGTCTCTGGTTTCTCATTAACTACCTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCT GGATTGGCTGGGAGTGATATGGAGTGGTGGAAGCACAGACTATAATGCAGCTTTCATATCCAGACTG AGCATCAGCAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAAACTAATGACA CAGCCATATATTACTGTGCCAGAAGAGGGAATGATGGTCCTCCGTTTGCTTACTGGGGCCAAGGGAC TCTGGTCACTGTCTCTGCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGAC ATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTC GGGCAAGTCAGGAAATTAGTGGTTACTTAAGCTGGCTTCAGAAGAAACCAGATGGAACTATTAAACG CCTGATCTACGCCGCATCCACTTTAGATTCTGGTGTCCCAAAAAGGTTCAGTGGCAGTAGGTCTGGG TCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGCAGACTATTACTGTCTACAAT ATGTTAGTTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 254) [00439] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 248 is set forth in SEQ ID NO: 255, which is provided below. CAAGTGCAGCTGAAGCAGAGCGGCCCTGGACTGGTGCAGCCTTCTCAGAGCCTCTCCATCACCTGCA CCGTGTCCGGCTTCAGCCTGACAACATACGGCGTGCACTGGGTGCGGCAGAGCCCCGGCAAGGGCCT GGACTGGCTGGGCGTGATCTGGTCCGGCGGCTCCACAGATTACAACGCCGCTTTTATCAGCAGACTG TCCATCAGCAAGGACAACAGCAAGAGCCAGGTGTTCTTCAAGATGAACAGCCTGCAGACCAATGACA CCGCCATCTACTACTGCGCCAGAAGGGGCAACGACGGCCCACCTTTCGCCTACTGGGGCCAGGGAAC ACTGGTGACCGTTAGTGCTGGCGGAGGCGGCAGCGGAGGAGGCGGCTCTGGCGGCGGCGGCTCAGAT ATCCAGATGACCCAGTCTCCTAGCAGCCTGAGCGCTAGCCTGGGCGAGAGAGTGTCCCTGACATGTA GAGCCAGCCAGGAGATCAGCGGCTACCTGAGCTGGCTGCAGAAAAAGCCCGACGGCACCATCAAGAG ACTGATCTACGCCGCCTCTACCCTGGATAGCGGCGTCCCCAAGCGGTTCAGCGGAAGCCGGAGCGGT TCTGATTACAGCCTGACCATTTCTAGCCTTGAAAGCGAGGACTTCGCCGACTACTATTGCCTGCAAT ACGTGTCTTATCCTCTGACTTTTGGAGCCGGCACCAAGCTGGAACTGAAA (SEQ ID NO: 255) [00440] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 249 is set forth in SEQ ID NO: 256, which is provided below. GACATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTT GTCGGGCAAGTCAGGAAATTAGTGGTTACTTAAGCTGGCTTCAGAAGAAACCAGATGGAACTATTAA ACGCCTGATCTACGCCGCATCCACTTTAGATTCTGGTGTCCCAAAAAGGTTCAGTGGCAGTAGGTCT NAI-1540294631v3 132 GGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGCAGACTATTACTGTCTAC AATATGTTAGTTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAAGGTGGAGGTGGATC AGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTG CAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTACCTATGGTGTAC ACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGATTGGCTGGGAGTGATATGGAGTGGTGGAAGCAC AGACTATAATGCAGCTTTCATATCCAGACTGAGCATCAGCAAGGACAATTCCAAGAGCCAAGTTTTC TTTAAAATGAACAGTCTGCAAACTAATGACACAGCCATATATTACTGTGCCAGAAGAGGGAATGATG GTCCTCCGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 256) [00441] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 249 is set forth in SEQ ID NO: 257, which is provided below. GATATCCAGATGACCCAGTCTCCTAGCAGCCTGAGCGCTAGCCTGGGCGAGAGAGTGTCCCTGACAT GTAGAGCCAGCCAGGAGATCAGCGGCTACCTGAGCTGGCTGCAGAAAAAGCCCGACGGCACCATCAA GAGACTGATCTACGCCGCCTCTACCCTGGATAGCGGCGTCCCCAAGCGGTTCAGCGGAAGCCGGAGC GGTTCTGATTACAGCCTGACCATTTCTAGCCTTGAAAGCGAGGACTTCGCCGACTACTATTGCCTGC AATACGTGTCTTATCCTCTGACTTTTGGAGCCGGCACCAAGCTGGAACTGAAAGGCGGAGGCGGCAG CGGAGGAGGCGGCTCTGGCGGCGGCGGCTCACAAGTGCAGCTGAAGCAGAGCGGCCCTGGACTGGTG CAGCCTTCTCAGAGCCTCTCCATCACCTGCACCGTGTCCGGCTTCAGCCTGACAACATACGGCGTGC ACTGGGTGCGGCAGAGCCCCGGCAAGGGCCTGGACTGGCTGGGCGTGATCTGGTCCGGCGGCTCCAC AGATTACAACGCCGCTTTTATCAGCAGACTGTCCATCAGCAAGGACAACAGCAAGAGCCAGGTGTTC TTCAAGATGAACAGCCTGCAGACCAATGACACCGCCATCTACTACTGCGCCAGAAGGGGCAACGACG GCCCACCTTTCGCCTACTGGGGCCAGGGAACACTGGTGACCGTTAGTGCT (SEQ ID NO: 257) [00442] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 262; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 263. SEQ ID NO: 262 and SEQ ID NO: 263 are disclosed in Table 16. [00443] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260. SEQ ID NOs: 258-260 are provided in Table 16. NAI-1540294631v3 133 [00444] In certain embodiments, the VH comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 263. For example, the VH comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 263. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 263. [00445] In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. SEQ ID NOs: 230, 231, and 261 are provided in Table 16. [00446] In certain embodiments, the VL comprises an amino acid sequence that is at least about 80% (e.g., at least about 85%, at least about 90%, or at least about 95%) identical or homologous to the amino acid sequence set forth in SEQ ID NO: 263. For example, the VL comprises an amino acid sequence that is about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 263. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 263. [00447] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof; and VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 NAI-1540294631v3 134 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. [00448] In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 262, and the VL comprises the amino acid sequence set forth in SEQ ID NO: 63. [00449] The VH the VL can be linked one after another, e.g., by a linker. In certain embodiments, the VH and VL are linked via a linker. In certain embodiments, the linker comprises or consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO : 4. The variable regions from the N- to the C-terminus can be VH-VL or VL-VH. [00450] In certain embodiments, the VH is positioned at the N-terminus of the extracellular domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VH-VL. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 262, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 263. In certain embodiments, the VH and VL are linked via a linker comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 264. SEQ ID NO: 264 is provided in Table 16. [00451] In certain embodiments, the VL is positioned at the N-terminus of the extracellular antigen-binding domain, i.e., the VH and the VL are positioned from the N- to the C-terminus as VL- VH. In certain embodiments, the extracellular domain of the CAR is an scFv that comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230; a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. In certain embodiments, the scFv comprises a VH comprising the amino acid sequence set forth NAI-1540294631v3 135 in SEQ ID NO: 262, and a VL comprising the amino acid sequence set forth in SEQ ID NO: 263. In certain embodiments, the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 265. SEQ ID NO: 265 is provided in Table 16. [00452] In certain embodiments, the CDRs are identified according to the Kabat numbering system. Table 16 (ATA0016 or ATA022) CDRs 1 2 3 VH NYWLH (SEQ ID NO: NIYPGYGTSNYDEKFTN GGYDYYFDY (SEQ Q Y Q H Y Q W Y H G Q R
Figure imgf000138_0001
[00453] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 262 is set forth in SEQ ID NO: 266, which is provided below. CAGGTCCAACTGCAGCAACCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCA AGGCTTCTGGCTACACATTCACCAACTACTGGTTGCACTGGGTGAAGCAAAGGCCTGGACAAGGCCT TGAGTGGATTGGAAATATTTATCCTGGTTATGGTACTTCTAACTACGATGAGAAGTTCACGAACAAG GCCGCACTGACTATAGACACATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGG ACTCTGCGGTCTATTTCTGTACAAGAGGGGGATATGATTACTACTTTGACTACTGGGGCCAAGGCAC CACTCTCACAGTCTCCTCA (SEQ ID NO: 266) [00454] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 262 is set forth in SEQ ID NO: 267, which is provided below. NAI-1540294631v3 136 CAAGTGCAGCTGCAGCAGCCTGGCAGCGAGCTGGTGCGGCCTGGCGCCAGCGTGAAACTGTCTTGTA AAGCTTCTGGCTACACCTTTACAAACTACTGGCTGCACTGGGTCAAGCAGAGACCTGGACAGGGCCT GGAATGGATCGGCAATATCTACCCTGGCTACGGCACAAGCAACTACGACGAGAAGTTCACCAACAAG GCCGCTCTGACAATCGATACCAGCAGCTCCACCGCCTACATGCAGCTGAGCAGCCTGACATCCGAGG ATTCTGCCGTGTACTTCTGCACCAGAGGCGGATATGATTACTACTTCGACTACTGGGGCCAGGGCAC CACCCTGACCGTGTCCAGC (SEQ ID NO: 267) [00455] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 263 is set forth in SEQ ID NO: 268, which is provided below. GACATTCAGATGAACCAGTCTCCATCCAGTCTGTCTGCATCCCTTGGAGACACAATTACCATCACTT GCCATGCCAGTCAGAACATTAATGTTTGGTTAAGCTGGTACCAGCAGACACCAGGAAATATTCCTAA ACTATTGATCTATAAGGCTTCCAACTTGCACACAGGCGTCCCATCAAGGTTTAGTGGCAGTGGATCT GGAACAGGTTTCACATTAACCATCAGCAGTCTGCAGCCTGAAGACATTGCCACTTACTACTGTCAAC AGGGTCAAAGTTATCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 268) [00456] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 263 is set forth in SEQ ID NO: 269, which is provided below. GACATCCAGATGAACCAGAGCCCTAGCTCCCTGAGCGCCAGCCTGGGCGACACCATTACCATCACCT GCCACGCCAGCCAGAACATCAACGTGTGGCTGAGCTGGTACCAGCAAACACCTGGCAACATCCCCAA GCTGCTGATCTATAAGGCCTCCAATCTGCACACAGGCGTGCCATCTAGATTCAGCGGCAGCGGCTCT GGAACCGGCTTTACCCTGACCATCAGCTCTCTCCAGCCCGAGGACATCGCCACATACTACTGCCAGC AGGGCCAAAGCTACCCCTGGACCTTCGGCGGCGGAACAAAGCTGGAAATCAAG (SEQ ID NO: 269) [00457] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 264 is set forth in SEQ ID NO: 270, which is provided below. CAGGTCCAACTGCAGCAACCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCA AGGCTTCTGGCTACACATTCACCAACTACTGGTTGCACTGGGTGAAGCAAAGGCCTGGACAAGGCCT TGAGTGGATTGGAAATATTTATCCTGGTTATGGTACTTCTAACTACGATGAGAAGTTCACGAACAAG GCCGCACTGACTATAGACACATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGG ACTCTGCGGTCTATTTCTGTACAAGAGGGGGATATGATTACTACTTTGACTACTGGGGCCAAGGCAC CACTCTCACAGTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGAC ATTCAGATGAACCAGTCTCCATCCAGTCTGTCTGCATCCCTTGGAGACACAATTACCATCACTTGCC ATGCCAGTCAGAACATTAATGTTTGGTTAAGCTGGTACCAGCAGACACCAGGAAATATTCCTAAACT ATTGATCTATAAGGCTTCCAACTTGCACACAGGCGTCCCATCAAGGTTTAGTGGCAGTGGATCTGGA NAI-1540294631v3 137 ACAGGTTTCACATTAACCATCAGCAGTCTGCAGCCTGAAGACATTGCCACTTACTACTGTCAACAGG GTCAAAGTTATCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 270) [00458] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 264 is set forth in SEQ ID NO: 271, which is provided below. CAAGTGCAGCTGCAGCAGCCTGGCAGCGAGCTGGTGCGGCCTGGCGCCAGCGTGAAACTGTCTTGTA AAGCTTCTGGCTACACCTTTACAAACTACTGGCTGCACTGGGTCAAGCAGAGACCTGGACAGGGCCT GGAATGGATCGGCAATATCTACCCTGGCTACGGCACAAGCAACTACGACGAGAAGTTCACCAACAAG GCCGCTCTGACAATCGATACCAGCAGCTCCACCGCCTACATGCAGCTGAGCAGCCTGACATCCGAGG ATTCTGCCGTGTACTTCTGCACCAGAGGCGGATATGATTACTACTTCGACTACTGGGGCCAGGGCAC CACCCTGACCGTGTCCAGCGGCGGCGGCGGCAGCGGAGGCGGCGGCAGCGGAGGAGGTGGCAGCGAC ATCCAGATGAACCAGAGCCCTAGCTCCCTGAGCGCCAGCCTGGGCGACACCATTACCATCACCTGCC ACGCCAGCCAGAACATCAACGTGTGGCTGAGCTGGTACCAGCAAACACCTGGCAACATCCCCAAGCT GCTGATCTATAAGGCCTCCAATCTGCACACAGGCGTGCCATCTAGATTCAGCGGCAGCGGCTCTGGA ACCGGCTTTACCCTGACCATCAGCTCTCTCCAGCCCGAGGACATCGCCACATACTACTGCCAGCAGG GCCAAAGCTACCCCTGGACCTTCGGCGGCGGAACAAAGCTGGAAATCAAG (SEQ ID NO: 271) [00459] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 265 is set forth in SEQ ID NO: 272, which is provided below. GACATTCAGATGAACCAGTCTCCATCCAGTCTGTCTGCATCCCTTGGAGACACAATTACCATCACTT GCCATGCCAGTCAGAACATTAATGTTTGGTTAAGCTGGTACCAGCAGACACCAGGAAATATTCCTAA ACTATTGATCTATAAGGCTTCCAACTTGCACACAGGCGTCCCATCAAGGTTTAGTGGCAGTGGATCT GGAACAGGTTTCACATTAACCATCAGCAGTCTGCAGCCTGAAGACATTGCCACTTACTACTGTCAAC AGGGTCAAAGTTATCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAAGGTGGAGGTGGATC AGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAACTGCAGCAACCTGGGTCTGAGCTGGTG AGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACATTCACCAACTACTGGTTGC ACTGGGTGAAGCAAAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTTATGGTAC TTCTAACTACGATGAGAAGTTCACGAACAAGGCCGCACTGACTATAGACACATCCTCCAGCACAGCC TACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTTCTGTACAAGAGGGGGATATG ATTACTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA (SEQ ID NO: 272) [00460] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 265 is set forth in SEQ ID NO: 273, which is provided below. GACATCCAGATGAACCAGAGCCCTAGCTCCCTGAGCGCCAGCCTGGGCGACACCATTACCATCACCT GCCACGCCAGCCAGAACATCAACGTGTGGCTGAGCTGGTACCAGCAAACACCTGGCAACATCCCCAA GCTGCTGATCTATAAGGCCTCCAATCTGCACACAGGCGTGCCATCTAGATTCAGCGGCAGCGGCTCT NAI-1540294631v3 138 GGAACCGGCTTTACCCTGACCATCAGCTCTCTCCAGCCCGAGGACATCGCCACATACTACTGCCAGC AGGGCCAAAGCTACCCCTGGACCTTCGGCGGCGGAACAAAGCTGGAAATCAAGGGCGGCGGCGGCAG CGGAGGCGGCGGCAGCGGAGGAGGTGGCAGCCAAGTGCAGCTGCAGCAGCCTGGCAGCGAGCTGGTG CGGCCTGGCGCCAGCGTGAAACTGTCTTGTAAAGCTTCTGGCTACACCTTTACAAACTACTGGCTGC ACTGGGTCAAGCAGAGACCTGGACAGGGCCTGGAATGGATCGGCAATATCTACCCTGGCTACGGCAC AAGCAACTACGACGAGAAGTTCACCAACAAGGCCGCTCTGACAATCGATACCAGCAGCTCCACCGCC TACATGCAGCTGAGCAGCCTGACATCCGAGGATTCTGCCGTGTACTTCTGCACCAGAGGCGGATATG ATTACTACTTCGACTACTGGGGCCAGGGCACCACCCTGACCGTGTCCAGC (SEQ ID NO: 273) [00461] The VH and/or VL amino acid sequences consisting of at least about 80%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% (e.g., about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99%) homology or identity to a specific sequence (e.g., SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, SEQ ID NO: 262, SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263) may contain substitutions (e.g., conservative substitutions), insertions, or deletions relative to the specified sequence(s), but retain the ability to bind to a target antigen (e.g., mesothelin). In certain embodiments, a total of 1 to 10 amino acids are substituted, inserted and/or deleted in a specific sequence (e.g., SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, SEQ ID NO: 262, SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263). In certain embodiments, substitutions, insertions, or deletions occur in regions outside the CDRs (e.g., in the FRs) of the extracellular antigen-binding domain. In certain embodiments, the extracellular domain comprises VH and/or VL sequence selected from SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NAI-1540294631v3 139 NO: 246, SEQ ID NO: 262, SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263, including post-translational modifications of SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, SEQ ID NO: 262, SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263. [00462] In addition, the extracellular domain can comprise a leader or a signal peptide that directs the nascent protein into the endoplasmic reticulum. Signal peptide or leader can be essential if the CAR is to be glycosylated and anchored in the cell membrane. The signal sequence or leader can be a peptide sequence (about 5, about 10, about 15, about 20, about 25, or about 30 amino acids long) present at the N-terminus of newly synthesized proteins that directs their entry to the secretory pathway. In certain embodiments, the signal peptide is covalently joined to the 5’ terminus (N- terminus) of the extracellular domain. In certain embodiments, the signal peptide comprises a CD8 polypeptide, e.g., the CAR comprises a truncated CD8 signal peptide. In certain embodiments, the signal peptide is generated from the antibody from it is derived. In certain embodiments, the signal peptide is linked to the N-terminus of a VH disclosed herein. In certain embodiments, the signal peptide is linked to the N-terminus of a VL disclosed herein. In certain embodiments, the signal peptide comprises or consists of the amino acid sequence set forth in SEQ ID NO: 274, SEQ ID NO: 275, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQ ID NO: 280, SEQ ID NO: 281, SEQ ID NO: 282, SEQ ID NO: 283, SEQ ID NO: 284, SEQ ID NO: 285, SEQ ID NO: 286, SEQ ID NO: 287, SEQ ID NO: 288, or SEQ ID NO: 289. MGWSCIIFFLVATATGVLS (SEQ ID NO: 274) MKCSWVIFFLMAVVTGVNS (SEQ ID NO: 275) MNFGLRLIFLVLVLKGVLC (SEQ ID NO: 276) MGWSCIILFLVATATGVHS (SEQ ID NO: 277) MGWSCIIFFLVATATGVHS (SEQ ID NO: 278) MGWSSIILFLVATASGVHS (SEQ ID NO: 279) MAVLGLLFCLVTFPSCVLS (SEQ ID NO: 280) MSPAQFLFLSVLWIRETNG (SEQ ID NO: 281) MSPAQFLFLLVLWIREIHG (SEQ ID NO: 282) NAI-1540294631v3 140 MDSQAQVLMLLLLWVSGTCG (SEQ ID NO: 283) MSPAQFLFLLVLWNRETNG (SEQ ID NO: 284) MESHTQAFVFAFLWLSGVDG (SEQ ID NO: 285) MSPAQFLFLLVLWIRETNG (SEQ ID NO: 286) MRVPAHVFGFLLLWFPGTRC (SEQ ID NO: 287) MSVLTQVLALLLLWLTGARC (SEQ ID NO: 288) MRVLAELLGLLLFCFLGVRC (SEQ ID NO: 289) [00463] In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 274 is set forth in SEQ ID NO: 290. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 275 is set forth in SEQ ID NO: 291. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 276 is set forth in SEQ ID NO: 292. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 277 is set forth in SEQ ID NO: 293 or SEQ ID NO: 294. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 278 is set forth in SEQ ID NO: 295. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 279 is set forth in SEQ ID NO: 296. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 280 is set forth in SEQ ID NO: 297. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 281 is set forth in SEQ ID NO: 298. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 282 is set forth in SEQ ID NO: 299. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 283 is set forth in SEQ ID NO: 300. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 284 is set forth in SEQ ID NO: 301. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 285 is set forth in SEQ ID NO: 302. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 286 is set forth in SEQ ID NO: 303 or SEQ ID NO: 304. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 287 is set forth in SEQ ID NO: 305. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 288 is set forth in SEQ ID NO: 306. In certain embodiments, an exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 289 is set forth in SEQ ID NO: 307. SEQ ID NOs: 290-307 are provided below. NAI-1540294631v3 141 ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCTCTCC (SEQ ID NO: 290) ATGAAATGCAGCTGGGTTATCTTCTTCCTGATGGCAGTGGTTACAGGGGTCAATTCA (SEQ ID NO: 291) ATGAACTTCGGGCTCAGATTGATTTTCCTTGTCCTTGTTTTAAAAGGTGTCCTGTGT (SEQ ID NO: 292) ATGGGTTGGAGCTGTATCATCCTTTTTCTGGTAGCAACAGCTACAGGTGTGCACTCC (SEQ ID NO: 293) ATGGGCTGGTCATGCATTATTCTGTTTCTGGTCGCAACTGCTACAGGCGTGCATAGT (SEQ ID NO: 294) ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCC (SEQ ID NO: 295) ATGGGATGGAGCTCTATCATCCTCTTCTTGGTAGCAACAGCCTCAGGTGTCCACTCC (SEQ ID NO: 296) ATGGCTGTCTTGGGGCTGCTCTTCTGCCTGGTGACATTCCCAAGCTGTGTCCTATCC (SEQ ID NO: 297) ATGAGTCCTGCCCAGTTCCTGTTTCTGTCAGTGCTCTGGATTCGGGAAACCAACGGT (SEQ ID NO: 298) ATGAGTCCTGCCCAGTTCCTGTTTCTGTTAGTGCTCTGGATTCGGGAAATCCACGGT (SEQ ID NO: 299) ATGGATTCACAGGCCCAGGTTCTTATGTTACTGCTGCTATGGGTATCTGGTACCTGTGGG (SEQ ID NO: 300) ATGAGTCCTGCCCAGTTCCTGTTTCTGTTAGTGCTCTGGAATCGGGAAACCAACGGT (SEQ ID NO: 301) ATGGAGTCTCATACTCAGGCCTTTGTATTCGCGTTTCTCTGGTTGTCTGGTGTTGATGGA (SEQ ID NO: 302) ATGAGTCCTGCCCAGTTCCTGTTTCTGTTAGTGCTCTGGATTCGGGAAACCAATGGT (SEQ ID NO: 303) ATGAGTCCTGCCCAGTTCCTGTTTCTGTTAGTGCTCTGGATTCGGGAAACCAACGGT (SEQ ID NO: 304) ATGAGGGTTCCTGCTCACGTTTTTGGCTTCTTGTTGCTCTGGTTTCCAGGTACCAGATGT (SEQ ID NO: 305) ATGAGTGTGCTCACTCAGGTCCTGGCGTTGCTGCTGCTGTGGCTTACAGGTGCCAGATGT (SEQ ID NO: 306) ATGAGGGTCCTTGCTGAGCTCCTGGGGCTGCTGCTGTTCTGCTTTTTAGGTGTGAGATGT (SEQ ID NO: 307) [00464] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 29 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00465] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 47 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00466] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 62 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00467] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75 and a signal peptide comprising the amino acid NAI-1540294631v3 142 sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00468] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 89 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 274. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00469] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 101 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00470] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00471] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00472] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 275. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00473] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 170 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00474] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00475] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201 and a signal peptide comprising the amino NAI-1540294631v3 143 acid sequence set forth in SEQ ID NO: 276. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00476] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 215 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00477] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 233 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 279. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00478] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 280. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00479] In certain embodiments, the extracellular domain of the CAR comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 262 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 279. In certain embodiments, the signal peptide is linked to the N-terminus of the VH. [00480] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 30, and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 281. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00481] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 48 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 286. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00482] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 63 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 286. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00483] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 76 and a signal peptide comprising the amino acid NAI-1540294631v3 144 sequence set forth in SEQ ID NO: 286. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00484] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 90 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 286. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00485] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 102 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 282. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00486] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 120 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 287. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00487] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 138 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 288. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00488] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 156 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 283. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00489] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 171 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 288. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00490] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 184 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 284. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00491] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 202 and a signal peptide comprising the amino NAI-1540294631v3 145 acid sequence set forth in SEQ ID NO: 285. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00492] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 216 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 288. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00493] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 234 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 289. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00494] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 247 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 287. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00495] In certain embodiments, the extracellular domain of the CAR comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 263 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 289. In certain embodiments, the signal peptide is linked to the N-terminus of the VL. [00496] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 31, and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00497] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 49 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00498] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 77 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00499] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 139 and a signal peptide comprising NAI-1540294631v3 146 the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00500] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 157 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00501] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 264 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00502] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 32, and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00503] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 50 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00504] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 78 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00505] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 140 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00506] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 158 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00507] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 265 and a signal peptide comprising NAI-1540294631v3 147 the amino acid sequence set forth in SEQ ID NO: 278. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00508] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 31, and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00509] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 49 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00510] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 64 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00511] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 77 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00512] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 91 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00513] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 103 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00514] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 121 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00515] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 139 and a signal peptide comprising NAI-1540294631v3 148 the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00516] In certain embodiments, the extracellular domain of the CAR f comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 157 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00517] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 172 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00518] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 185 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00519] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 203 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00520] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 217 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00521] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 235 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00522] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 248 and a signal peptide comprising the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. [00523] In certain embodiments, the extracellular domain of the CAR comprises an scFv comprising the amino acid sequence set forth in SEQ ID NO: 264 and a signal peptide comprising NAI-1540294631v3 149 the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the signal peptide is linked to the N-terminus of the scFv. 5.3.2. Transmembrane Domain of a CAR [00524] In certain embodiments, the transmembrane domain of the CAR comprises a hydrophobic alpha helix that spans at least a portion of the membrane. Different transmembrane domains result in different receptor stability. After antigen recognition, receptors cluster and a signal are transmitted to the cell. [00525] The transmembrane domain may be derived either from a natural or from a synthetic source. Where the source is natural, the domain may be derived from any membrane-bound or transmembrane protein. For example, the transmembrane region may be derived from (e.g., comprise at least the transmembrane region(s) of) the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8 (e.g., CD8 alpha, CD8 beta), CD9, CD16, CD22, CD27, CD33, CD37, CD40, CD64, CD80, CD86, CD134, CD137, CD154, ICAM-1, KIRDS2, OX40, CD2, NKGD2, LFA-1 (CD11a, CD18) , ICOS (CD278), 4-1BB (CD137), GITR, CD40, BAFFR, HVEM (LIGHTR) , SLAMF7, NKp80 (KLRF1), CD160, CD166, CD19, IL2R beta, IL2R gamma, IL7R α, ITGA1, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, ITGB7, TNFR2, DNAM1 (CD226) , SLAMF4 (CD244, 2B4) , CD84, CD96 (Tactile) , CEACAM1, CRTAM, Ly9 (CD229) , CD160 (BY55) , PSGL1, CD100 (SEMA4D), SLAMF6 (NTB-A, Ly108) , SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, and PAG/Cbp. In certain embodiments, the transmembrane domain of the CAR comprises a native or modified transmembrane domain of CD8, of CD28, of CD3ζ, of CD4, of 4-1BB, of OX40, of ICOS, of CD84, of CD166, of CD8a, of CD8b, of ICAM-1, of CTLA-4, of CD27, of CD40, of NKGD2, or a combination thereof. Alternatively, the transmembrane domain may be synthetic, in which case it comprises predominantly hydrophobic residues such as leucine and valine. In some cases, a triplet of phenylalanine, tryptophan and valine can be found at each end of a synthetic transmembrane domain. A short oligo- or polypeptide linker, such as between 2 and 10 amino acids in length, may form the linkage between the transmembrane domain and the intracellular domain of the CAR. [00526] In certain embodiments, the CAR comprises more than one transmembrane domain, which can be a repeat of the same transmembrane domain, or can be different transmembrane domains. [00527] In certain embodiments, the transmembrane domain of the CAR comprises a CD28 polypeptide (e.g., a transmembrane domain of CD28 or a portion thereof). In certain embodiments, NAI-1540294631v3 150 the transmembrane domain of the CAR comprises a transmembrane domain of human CD28 or a portion thereof. The CD28 polypeptide can comprise or consist of an amino acid sequence that is at least about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99% or 100% identical or homologous to the sequence having a NCBI Reference No: NP_006130 (SEQ ID NO: 308), or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions. In certain embodiments, the CD28 polypeptide comprises or consists of an amino acid sequence that is a consecutive portion of SEQ ID NO: 308, which is at least about 20, or at least about 30, or at least about 40, or at least about 50, or at least about 60, and/or up to about 70, up to about 80, up to about 90, up to about 100, up to about 150, up to about 200, or up to about 220 amino acids in length. In certain embodiments, the CD28 polypeptide comprises or consists of the amino acid sequence of amino acids 1 to 220, 1 to 50, 50 to 100, 100 to 150, 154 to 179, 150 to 200, 153 to 179, or 200 to 220 of SEQ ID NO: 308. In certain embodiments, the transmembrane domain of the CAR comprises a CD28 polypeptide comprising or consisting of the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308. SEQ ID NO: 308 is provided below. MLRLLLALNLFPSIQVTGNKILVKQSPMLVAYDNAVNLSCKYSYNLFSREFRASLHKGLDSAVEVCV VYGNYSQQLQVYSKTGFNCDGKLGNESVTFYLQNLYVNQTDIYFCKIEVMYPPPYLDNEKSNGTIIH VKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGP TRKHYQPYAPPRDFAAYRS [SEQ ID NO: 308] [00528] An exemplary nucleotide sequence encoding the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308 is set forth in SEQ ID NO: 309, which is provided below. TGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTA TTTTCTGGGTG [SEQ ID NO: 309] [00529] In certain embodiments, the transmembrane domain of the CAR comprises a CD8 polypeptide (e.g., a transmembrane domain of CD8 or a portion thereof). In certain embodiments, the transmembrane domain of the CAR comprises a transmembrane domain of human CD8 or a portion thereof. In certain embodiments, the CD8 polypeptide comprises or has an amino acid sequence that is at least about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the sequence having a NCBI Reference No: NP_001139345.1 (SEQ ID NO: 310) or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions. In certain embodiments, the CD8 polypeptide comprises or consists of an amino acid sequence that is a consecutive portion of SEQ ID NO: 310, which is at least about 20, or at least about 30, or at least about 40, or at least NAI-1540294631v3 151 about 50, and up to about 235 amino acids in length. Alternatively or additionally, in certain embodiments, the CD8 polypeptide comprises or consists of an amino acid sequence of amino acids 1 to 235, 1 to 50, 50 to 100, 100 to 150, 150 to 200, 137 to 209 or 200 to 235 of SEQ ID NO: 310. In certain embodiments, the transmembrane domain of the CAR comprises a CD8 polypeptide comprising or consisting of the amino acid sequence of amino acids 137 to 209 of SEQ ID NO: 310. SEQ ID NO: 310 is provided below. MALPVTALLLPLALLLHAARPSQFRVSPLDRTWNLGETVELKCQVLLSNPTSGCSWLFQPRGAAASP TFLLYLSQNKPKAAEGLDTQRFSGKRLGDTFVLTLSDFRRENEGYYFCSALSNSIMYFSHFVPVFLP AKPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLV ITLYCNHRNRRRVCKCPRPVVKSGDKPSLSARYV [SEQ ID NO: 310] [00530] In certain embodiments, the CAR further comprise a spacer/hinge domain that links the extracellular domain to the transmembrane domain. A hinge domain is a short sequence of amino acids that facilitates antibody flexibility (see, e.g., Woof et al., Nat. Rev. Immunol.4(2): 89-99 (2004)). The hinge domain can be any suitable sequence derived or obtained from any suitable molecule. The hinge domain can be flexible enough to allow the extracellular domain to orient in different directions to facilitate antigen recognition while preserving the activating activity of the CAR. [00531] In some embodiments, for example, the hinge domain sequence is derived from a CD8a molecule or a CD28 molecule. [00532] In certain embodiments, the hinge/spacer domain of the CAR comprises a native or modified hinge region of CD8, of CD28, of CD3ζ, of CD40, of 4-1BB, of OX40, of CD84, of CD166, of CD8a, of CD8b, of ICOS, of ICAM-1, of CTLA-4, of CD27, of CD40, of NKGD2, or a combination thereof. The hinge/spacer region can be the hinge region from IgG1, or the CH2CH3 region of immunoglobulin and portions of CD3, a portion of a CD28 polypeptide (e.g., a portion of SEQ ID NO: 308), a portion of a CD8 polypeptide (e.g., a portion of SEQ ID NO: 310), a variant of any of the foregoing which is at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100% identical or homologous thereto, or a synthetic spacer sequence. [00533] In certain embodiments, the hinge domain of the CAR comprises a native or modified hinge region of CD28. In certain embodiments, the hinge domain of the CAR comprises a native hinge region of CD28. In certain embodiments, the hinge domain of the CAR comprises the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308. An exemplary nucleotide sequence encoding the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308 is set forth in SEQ ID NO: 311, which is provided below. NAI-1540294631v3 152 ATTGAAGTTATGTATCCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGA AAGGGAAACACCTTTGTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTT [SEQ ID NO: 311] 5.3.3. Intracellular Domain of a CAR [00534] In certain embodiments, the CAR comprises an intracellular domain. In certain embodiments, the intracellular domain of the CAR comprises a CD3ζ polypeptide. CD3ζ can activate or stimulate a cell (e.g., an immune effector cell, e.g., a cell of the lymphoid lineage, e.g., a T-cell) after antigen recognition. The intracellular domain of the CD3ζ-chain is the primary transmitter of signals from endogenous TCRs. [00535] In certain embodiments, the intracellular domain of the CAR transmits a signal to the immune effector cell expressing the CAR after antigen recognition, activating at least one of the normal effector functions of the immune effector cell. In certain embodiments, the effector function of a T cell, for example, is cytolytic activity or helper activity, including secretion of cytokines. Therefore, the intracellular domain can comprise the “intracellular signaling domain” of a T cell receptor (TCR) and optional co-receptors. While usually the entire intracellular signaling domain can be employed, in many cases it is not necessary to use the entire chain. To the extent that a truncated portion of the intracellular signaling domain is used, such truncated portion may be used in place of the intact chain as long as it transduces the effector function signal. [00536] Cytoplasmic signaling sequences that regulate primary activation of the TCR complex that act in a stimulatory manner may contain signaling motifs which are known as immunoreceptor tyrosine-based activation motifs (ITAMs). Examples of ITAM containing cytoplasmic signaling sequences include those derived from CD8, CD3ζ, CD3δ, CD3γ, CD3ε, CD32 (Fc gamma RIIa), DAP10, DAP12, CD79a, CD79b, FcγRIγ, FcγRIIIγ, FcεRIβ (FCERIB), and FcεRIγ (FCERIG). [00537] In certain embodiments, the intracellular signaling domain is derived from CD3 zeta (CD3ζ) (TCR zeta, GenBank acc. no. BAG36664.1). T-cell surface glycoprotein CD3 zeta (CD3ζ) chain, also known as T-cell receptor T3 zeta chain or CD247 (Cluster of Differentiation 247), is a protein that in humans is encoded by the CD247 gene. Wild type (“native”) CD3ζ comprises three functional immunoreceptor tyrosine-based activation motifs (ITAMs), three functional basic-rich stretch (BRS) regions (BRS1, BRS2 and BRS3). The intracellular tails of the CD3 molecules comprise a single ITAM, which is responsible for the signaling capacity of the TCR. The intracellular tail of the ζ chain (CD3ζ) comprises three (3) ITAMs. [00538] In certain embodiments, the intracellular domain of the CAR comprises a native CD3ζ. In certain embodiments, the native CD3ζ comprises or consists of an amino acid sequence that is at NAI-1540294631v3 153 least about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99% or about 100% identical or homologous to the amino acid sequence having a NCBI Reference No: NP_932170 (SEQ ID NO: 312), or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions. In certain embodiments, the CD3ζ polypeptide comprises or consists of an amino acid sequence that is a consecutive portion of SEQ ID NO: 312, which is at least about 20, or at least about 30, or at least about 40, or at least about 50, and up to about 164 amino acids in length. In certain embodiments, the native CD3ζ comprises or consists of the amino acid sequence of amino acids 1 to 164, 1 to 50, 50 to 100, 52 to 164, 100 to 150, or 150 to 164 of SEQ ID NO: 312. In certain embodiments, the intracellular signaling domain of the CAR comprises a native CD3ζ comprising or consisting of the amino acid sequence of amino acids 52 to 164 of SEQ ID NO: 312. SEQ ID NO: 312 is provided below. MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQG QNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRR GKGHDGLYQGLSTATKDTYDALHMQALPPR [SEQ ID NO: 312] [00539] In certain embodiments, the intracellular domain of the CAR comprises a modified CD3ζ polypeptide. In certain embodiments, the modified CD3ζ polypeptide comprises a mutation, such as a point mutation, in at least one ITAM so as to render the ITAM non-functional. In certain embodiments, either the membrane-proximal ITAM (ITAM1), the membrane-distal ITAM (C- terminal third ITAM, ITAM3), or both are non-functional. In certain embodiments, either two membrane-proximal ITAMs (ITAM1 and ITAM2) or two membrane-distal ITAMs (ITAM2 and ITAM3) are non-functional. In certain embodiments, only ITAM2 is non-functional. In certain embodiments, the modified CD3ζ polypeptide comprises a deletion (e.g., a truncation) mutation such that at least one ITAM is missing. In certain embodiments, the modified CD3ζ polypeptide is missing the membrane-proximal ITAM (ITAM1), the membrane-distal ITAM (ITAM3), or both. In other embodiments, the modified CD3ζ polypeptide is missing either two membrane-proximal ITAMs (ITAM1 and ITAM2) or two membrane-distal ITAMs (ITAM2 and ITAM3). In certain embodiments, the modified CD3ζ polypeptide is missing ITAM2. Removing at least one ITAM from the introduced CAR may reduce CD3ζ-mediated apoptosis. Alternatively, removing at least one ITAM from the introduced CAR can reduce its size without loss of function. [00540] In certain embodiments, the modified CD3ζ polypeptide comprises one, two or three ITAMs. In certain embodiments, the modified CD3ζ polypeptide comprises a native ITAM1. In certain embodiments, the native ITAM1 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 313. NAI-1540294631v3 154 QNQLYNELNLGRREEYDVLDKR [SEQ ID NO: 313] [00541] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 313 is set forth in SEQ ID NO: 314, which is provided below. CAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGA [SEQ ID NO: 314] [00542] In certain embodiments, the modified CD3ζ polypeptide comprises an ITAM1 variant comprising one or more loss-of-function mutations. In certain embodiments, the ITAM1 variant comprises or consists of two loss-of-function mutations. In certain embodiments, each of the one or more (e.g., two) loss of function mutations comprises a mutation of a tyrosine residue in ITAM1. In certain embodiments, the ITAM1 variant consists of two loss-of-function mutations. In certain embodiments, the ITAM1 variant comprises or consists of the amino acid sequence set forth in SEQ ID NO: 315, which is provided below. QNQLFNELNLGRREEFDVLDKR [SEQ ID NO: 315] [00543] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 315 is set forth in SEQ ID NO: 316, which is provided below. CAGAACCAGCTCTTTAACGAGCTCAATCTAGGACGAAGAGAGGAGTTCGATGTTTTGGACAAGAGA [SEQ ID NO: 316] [00544] In certain embodiments, the modified CD3ζ polypeptide comprises a native ITAM2. In certain embodiments, the native ITAM2 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 317, which is provided below. QEGLYNELQKDKMAEAYSEIGMK [SEQ ID NO: 317] [00545] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 317 is set forth in SEQ ID NO: 318, which is provided below. CAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGA AA [SEQ ID NO: 318] [00546] In certain embodiments, the modified CD3ζ polypeptide comprises an ITAM2 variant. In certain embodiments, the ITAM2 variant comprises or consists of one or more loss-of-function mutations. In certain embodiments, the ITAM2 variant comprises or consists of two loss-of-function mutations. In certain embodiments, each of the one or more (e.g., two) the loss of function mutations comprises a mutation of a tyrosine residue in ITAM2. In certain embodiments, the ITAM1 variant consists of two loss-of-function mutations. In certain embodiments, the ITAM2 variant comprises or consists of the amino acid sequence set forth in SEQ ID NO: 319, which is provided below. QEGLFNELQKDKMAEAFSEIGMK [SEQ ID NO: 319] NAI-1540294631v3 155 [00547] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 319 is set forth in SEQ ID NO: 320, which is provided below. CAGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAGTGAGATTGGGATGA AA [SEQ ID NO: 320] [00548] In certain embodiments, the modified CD3ζ polypeptide comprises a native ITAM3. In certain embodiments, the native ITAM3 comprises or consists of the amino acid sequence set forth in SEQ ID NO: 321, which is provided below. HDGLYQGLSTATKDTYDALHMQ [SEQ ID NO: 321] [00549] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 321 is set forth in SEQ ID NO: 322, which is provided below. CACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAG [SEQ ID NO: 322] [00550] In certain embodiments, the modified CD3ζ polypeptide comprises an ITAM3 variant. In certain embodiments, the ITAM3 variant comprises or consists of two loss-of-function mutations. In certain embodiments, each of the one or more (e.g., two) the loss of function mutations comprises a mutation of a tyrosine residue in ITAM3. In certain embodiments, the ITAM3 variant comprises or consists of two loss-of-function mutations. In certain embodiments, the ITAM3 variant comprises or consists of the amino acid sequence set forth in SEQ ID NO: 323, which is provided below. HDGLFQGLSTATKDTFDALHMQ [SEQ ID NO: 323] [00551] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 323 is set forth in SEQ ID NO: 324, which is provided below. CACGATGGCCTTTTCCAGGGGCTCAGTACAGCCACCAAGGACACCTTCGACGCCCTTCACATGCAG [SEQ ID NO: 324] [00552] Various modified CD3ζ polypeptides and CARs comprising modified CD3ζ polypeptides are disclosed in International Patent Application Publication No. WO2019/133969, which is incorporated by reference hereby in its entirety. [00553] In certain embodiments, the intracellular domain of the CAR comprises a modified CD3ζ polypeptide comprising a native ITAM1, an ITAM2 variant comprising or consisting of one or more (e.g., two) loss-of-function mutations, and an ITAM3 variant comprising or consisting of one or more (e.g., two) loss-of-function mutations. In certain embodiments, the intracellular domain of the CAR comprises a modified CD3ζ polypeptide comprising a native ITAM1, an ITAM2 variant consisting of two loss-of-function mutations, and an ITAM3 variant consisting of two loss-of- function mutations. In certain embodiments, the intracellular domain of the CAR comprises a NAI-1540294631v3 156 modified CD3ζ polypeptide comprising a native ITAM1 consisting of the amino acid sequence set forth in SEQ ID NO: 313, an ITAM2 variant consisting of the amino acid sequence set forth in SEQ ID NO: 319, and an ITAM3 variant consisting of the amino acid sequence set forth in SEQ ID NO: 323. In certain embodiments, the modified CD3ζ polypeptide comprises or consists of the amino acid sequence set forth in SEQ ID NO: 325. SEQ ID NO: 325 is provided below. In certain embodiments, a CAR comprising an intracellular domain comprising a modified CD3ζ polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 325 is designated as an “1XX” CAR. RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKM AEAFSEIGMKGERRRGKGHDGLFQGLSTATKDTFDALHMQALPPR [SEQ ID NO: 325] [00554] In certain embodiments, the intracellular domain of the CAR comprises a modified CD3ζ polypeptide comprising or consisting of an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, at least about 100% identical to SEQ ID NO: 43 or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions. [00555] An exemplary nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 325 is set forth in SEQ ID NO: 326, which is provided below. AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACG AGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGAT GGGGGGAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATG GCGGAGGCCTTCAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTT TCCAGGGGCTCAGTACAGCCACCAAGGACACCTTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCG C [SEQ ID NO: 326] [00556] In certain embodiments, the intracellular domain of the CAR further comprises at least one co-stimulatory signaling region. In certain embodiments, the at least one co-stimulatory region comprises a co-stimulatory molecule or a portion thereof. In certain embodiments, the at least one co-stimulatory region comprises an intracellular domain of at least one co-stimulatory molecule or a portion thereof. [00557] As used herein, a “co-stimulatory molecule” refers to a cell surface molecule other than antigen receptor or its ligand that can provide an efficient response of lymphocytes to an antigen. A co-stimulatory molecule can provide optimal lymphocyte activation. The co-stimulatory molecule can bind to a co-stimulatory ligand, which is a protein expressed on cell surface that upon binding to its receptor produces a co-stimulatory response, i.e., an intracellular response that effects the NAI-1540294631v3 157 stimulation provided when an antigen-recognizing receptor (e.g., a chimeric antigen receptor (CAR)) binds to its target antigen. As one example, a 4-1BB ligand (i.e., 4-1BBL) may bind to 4-1BB for providing an intracellular signal that in combination with a CAR signal induces an effector cell function of the CAR+ T-cell. [00558] Non-limiting examples of co-stimulatory molecules include CD28, 4-1BB (CD137), OX40, ICOS, DAP-10, CD27, CD28, CD30, CD40, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, CD8, CD4, B2C, CD80, CD86, DAP10, DAP12, MyD88, BTNL3, and NKG2D. and combinations thereof. [00559] In certain embodiments, the intracellular domain of the CAR comprises a co-stimulatory signaling region that comprises a CD28 polypeptide, e.g., an intracellular domain of CD28 or a portion thereof. In certain embodiments, the intracellular domain of the CAR comprises a co- stimulatory signaling region that comprises an intracellular domain of human CD28 or a portion thereof. [00560] In certain embodiments, the CD28 polypeptide comprised in the co-stimulatory signaling region of the CAR comprise or consists of an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, at least about 100% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 308, or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions. In certain embodiments, the CD28 polypeptide comprised in the co-stimulatory signaling region comprises or consist of an amino acid sequence that is a consecutive portion of SEQ ID NO: 308, which is at least about 20, or at least about 30, or at least about 40, or at least about 50, and up to about 220 amino acids in length. Alternatively or additionally, in certain embodiments, the CD28 polypeptide comprised in the co- stimulatory signaling region comprises or consists of amino acids 1 to 220, 1 to 50, 50 to 100, 100 to 150, 114 to 220, 150 to 200, 180 to 220, or 200 to 220 of SEQ ID NO: 308. In certain embodiments, the intracellular signaling domain of the CAR comprises a co-stimulatory signaling region that comprises a CD28 polypeptide comprising or consisting of the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308. [00561] An exemplary nucleic acid sequence encoding the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308 is set forth in SEQ ID NO: 327, which is provided below. NAI-1540294631v3 158 AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCA CCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCC [SEQ ID NO: 327] [00562] In certain embodiments, the intracellular domain of the CAR comprises a modified CD28 domain that imparts unique functional properties to the CAR. In this regard, a native CD28 domain comprises three intracellular subdomains consisting of the amino acid sequences YMNM, PRRP, and PYAP that regulate signaling pathways post stimulation. In certain embodiments, the intracellular domain of the CAR comprises a modified CD28 domain wherein one or more of the YMNM, PRRP, and PYAP subdomains are mutated or deleted, so as to amplify, attenuate, or inactivate said subdomain(s), thereby modulating CAR-T function. In certain embodiments, the intracellular domain of the CAR comprises a modified CD28 domain disclosed in International Patent Application Publication Nos.: WO 2019/010383 and WO 2021158850, which are incorporated by reference in their entireties. [00563] In certain embodiments, the intracellular domain of the CAR comprises a co-stimulatory signaling region that comprises a 4-1BB polypeptide, e.g., an intracellular domain of 4-1BB or a portion thereof. In certain embodiments, the co-stimulatory signaling region comprises an intracellular domain of human 4-1BB or a portion thereof. In certain embodiments, the 4-1BB comprised in the co-stimulatory signaling region comprises or consists of an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%, at least about 100% identical or homologous to the sequence having a NCBI Ref No.: NP_001552 (SEQ ID NO: 328) or a fragment thereof, and/or may optionally comprise up to one or up to two or up to three conservative amino acid substitutions. In certain embodiments, the 4-1BB comprised in the co-stimulatory signaling region comprises or consists of an amino acid sequence that is a consecutive portion of SEQ ID NO: 328, which is at least about 20, or at least about 30, or at least about 40, or at least about 50, and/or up to about 50, up to about 60, up to about 70, up to about 80, up to about 90, up to about 100, up to about 200, or up to about 255 amino acids in length. In certain embodiments, the 4-1BB polypeptide comprised in the co-stimulatory signaling region comprises or consists of the amino acid sequence of amino acids 1 to 255, 1 to 50, 50 to 100, 100 to 150, 150 to 200, or 200 to 255 of SEQ ID NO: 328. In certain embodiments, the co-stimulatory signaling region comprises a 4-1BB polypeptide comprising or consisting of the amino acid sequence of amino acids 214 to 255 of SEQ ID NO: 328. SEQ ID NO: 328 is provided below. NAI-1540294631v3 159 MGNSCYNIVATLLLVLNFERTRSLQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQ CKGVFRTRKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQELTKKGCKDCCFGTFNDQKRGICR PWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSVTPPAPAREPGHSPQIISFFLALTSTALLF LLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL [SEQ ID NO: 328] [00564] In certain embodiments, the intracellular domain of the CAR comprises a co-stimulatory signaling region that comprises intracellular domains of two or more co-stimulatory molecules or portions thereof, e.g., an intracellular domain of CD28 or a portion thereof and an intracellular domain of 4-1BB or a portion thereof, or an intracellular domains of CD28 or a portion thereof and an intracellular domain of OX40 or a portion thereof. [00565] In certain embodiments, the intracellular domain of the CAR further comprises a peptide. In certain embodiments, the self-cleaving P2A peptide comprises or consists of the amino acid sequence of SEQ ID NO: 329. SEQ ID NO: 329 is provided below. GSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 329] [00566] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 329 is set forth in SEQ ID NO: 330, which is provided below. GGATCTGGAGCAACAAACTTCTCACTACTCAAACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 330] [00567] In certain embodiments, the CAR is expressed with a C-terminal LNGFR tag. In certain embodiments, the LNGFR tag comprises or consists of the amino acid sequence of SEQ ID NO: 331. SEQ ID NO: 331 is provided below. MGAGATGRAMDGPRLLLLLLLGVSLGGAKEACPTGLYTHSGECCKACNLGEGVAQPCGANQTVCEPC LDSVTFSDVVSATEPCKPCTECVGLQSMSAPCVEADDAVCRCAYGYYQDETTGRCEACRVCEAGSGR SSGLVFSCQDKQNTVCEECPDGTYSDEANHVDPCLPCTVCEDTERQLRECTRWADAECEEIPGRWIT RSTPPEGSDSTAPSTQEPEAPPEQDLIASTVAGVVTTVMGSSQPVVTRGTTDNLIPVYCSILAAVVV GLVAYIAFKRWNS [SEQ ID NO: 331] [00568] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 331 is set forth in SEQ ID NO: 332, which is provided below. ATGGGGGCAGGGGCCACCGGACGCGCAATGGACGGGCCGCGCCTGCTGCTGTTGCTGCTTCTGGGGG TGTCCCTTGGAGGTGCCAAGGAGGCATGCCCCACAGGCCTGTACACACACAGCGGTGAGTGCTGCAA AGCCTGCAACCTGGGCGAGGGTGTGGCCCAGCCTTGTGGAGCCAACCAGACCGTGTGTGAGCCCTGC CTGGACAGCGTGACGTTCTCCGACGTGGTGAGCGCGACCGAGCCGTGCAAGCCGTGCACCGAGTGCG TGGGGCTCCAGAGCATGTCGGCGCCGTGCGTGGAGGCCGACGACGCCGTGTGCCGCTGCGCCTACGG NAI-1540294631v3 160 CTACTACCAGGATGAGACGACTGGGCGCTGCGAGGCGTGCCGCGTGTGCGAGGCGGGCTCGGGCCGC TCGAGCGGCCTCGTGTTCTCCTGCCAGGACAAGCAGAACACCGTGTGCGAGGAGTGCCCCGACGGCA CGTATTCCGACGAGGCCAACCACGTGGACCCGTGCCTGCCCTGCACCGTGTGCGAGGACACCGAGCG CCAGCTCCGCGAGTGCACACGCTGGGCCGACGCCGAGTGCGAGGAGATCCCTGGCCGTTGGATTACA CGGTCCACACCCCCAGAGGGCTCGGACAGCACAGCCCCCAGCACCCAGGAGCCTGAGGCACCTCCAG AACAAGACCTCATAGCCAGCACGGTGGCAGGTGTGGTGACCACAGTGATGGGCAGCTCCCAGCCCGT GGTGACCCGAGGCACCACCGACAACCTCATCCCTGTCTATTGCTCCATCCTGGCTGCTGTGGTTGTG GGCCTTGTGGCCTACATAGCCTTCAAGAGGTGGAACAGCTGA [SEQ ID NO: 332] 5.3.4. Exemplified CARs [00569] In certain embodiments, the CAR comprises (a) an extracellular antigen-binding domain comprising (i) a VH that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 25, and (ii) a VL that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 28, (b) a hinge domain comprising a CD28 polypeptide (e.g., a hinge domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308); (c) a transmembrane domain comprising a CD28 polypeptide (e.g., a transmembrane domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308), and (d) an intracellular signaling domain comprising (i) a CD3ζ polypeptide (e.g., a modified human CD3ζ polypeptide, e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 325), and (ii) a co-stimulatory signaling region comprising a CD28 polypeptide (e.g., an intracellular domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308). In certain embodiments, the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329. In certain embodiments, the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278). In certain embodiments, the VH and VL are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00570] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VH- VL. In certain embodiments, the CAR is designed as “1_28z1XX (vH-vK)” or “ATA017 VH-VK CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 333, which is provided below. NAI-1540294631v3 161 MGWSCIIFFLVATATGVHSQVQLQQSGPEVVRPGVSVKISCKGSGYKFTDYAMHWVKQSHAKSLEWI GVISTYNGNTNYNRKFKDKATMTVDKSSSTAYMELARLTSEDSAIYYCLKNDYWGQGTSVTVSSGGG GSGGGGSGGGGSDVVMTQTPLTLSVTIGQPASFSCKSSQSLLDSDGKTYLNWVLQRPGQSPNRLIYL VSKLDSRVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTQFPHTFGGGTKLEIKRAAAIEVMYP PPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRL LHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYD VLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLSTATKDT FDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 333] [00571] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 333 is set forth in SEQ ID NO: 334, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCCAGGTCCAGC TGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGG CTACAAATTCACTGATTATGCTATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATT GGAGTTATTAGTACTTACAATGGTAATACAAACTACAACCGGAAGTTTAAGGACAAGGCCACAATGA CTGTAGACAAATCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAAGATTCTGCCAT CTATTACTGTTTAAAGAACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGT GGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACCCAGACTCCACTCACTT TGTCGGTTACCATTGGACAACCAGCCTCCTTCTCTTGCAAGTCAAGTCAGAGTCTCTTAGATAGTGA TGGAAAGACATATTTGAATTGGGTGTTACAGAGGCCAGGCCAGTCGCCAAATCGCCTAATCTATCTG GTGTCTAAATTGGACTCTAGAGTCCCTGACAGGTTCACTGGCAGTGGATCAGGGACAGATTTCACAC TGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGCAAGGTACACAGTTTCC TCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAACGGGCGGCCGCAATTGAAGTTATGTATCCT CCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGAAAGGGAAACACCTTTGTC CAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGC TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGGAGCAGGCTC CTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCT ATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCC CGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGAT GTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGG AAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAGTGAGATTGGGATGAAAGG CGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGTACAGCCACCAAGGACACC TTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAACAAACTTCTCACTACTCA AACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 334] NAI-1540294631v3 162 [00572] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VL- VH. In certain embodiments, the CAR is designed as “1_28z1XX (vK-vH)” or “ATA017 VK-VH CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 335, which is provided below. MGWSCIIFFLVATATGVHSDVVMTQTPLTLSVTIGQPASFSCKSSQSLLDSDGKTYLNWVLQRPGQS PNRLIYLVSKLDSRVPDRFTGSGSGTDFTLKISRVEAEDLGVYYCWQGTQFPHTFGGGTKLEIKGGG GSGGGGSGGGGSQVQLQQSGPEVVRPGVSVKISCKGSGYKFTDYAMHWVKQSHAKSLEWIGVISTYN GNTNYNRKFKDKATMTVDKSSSTAYMELARLTSEDSAIYYCLKNDYWGQGTSVTVSSRAAAIEVMYP PPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRL LHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYD VLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLSTATKDT FDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 335] [00573] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 335 is set forth in SEQ ID NO: 336, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCGATGTTGTGA TGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCTTCTCTTGCAAGTCAAG TCAGAGTCTCTTAGATAGTGATGGAAAGACATATTTGAATTGGGTGTTACAGAGGCCAGGCCAGTCG CCAAATCGCCTAATCTATCTGGTGTCTAAATTGGACTCTAGAGTCCCTGACAGGTTCACTGGCAGTG GATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTG CTGGCAAGGTACACAGTTTCCTCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAAGGTGGAGGT GGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAGCTGCAGCAGTCTGGGCCTGAGG TGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACAAATTCACTGATTATGC TATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATTAGTACTTACAAT GGTAATACAAACTACAACCGGAAGTTTAAGGACAAGGCCACAATGACTGTAGACAAATCCTCCAGCA CAGCCTATATGGAACTTGCCAGATTGACATCTGAAGATTCTGCCATCTATTACTGTTTAAAGAACGA CTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCACGGGCGGCCGCAATTGAAGTTATGTATCCT CCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGAAAGGGAAACACCTTTGTC CAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGC TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGGAGCAGGCTC CTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCT ATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCC CGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGAT GTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGG NAI-1540294631v3 163 AAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAGTGAGATTGGGATGAAAGG CGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGTACAGCCACCAAGGACACC TTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAACAAACTTCTCACTACTCA AACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 336] [00574] In certain embodiments, the CAR comprises (a) an extracellular antigen-binding domain comprising (i) a VH that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 43, and (ii) a VL that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 46, (b) a hinge domain comprising a CD28 polypeptide (e.g., a hinge domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308); (c) a transmembrane domain comprising a CD28 polypeptide (e.g., a transmembrane domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308), and (d) an intracellular signaling domain comprising (i) a CD3ζ polypeptide (e.g., a modified human CD3ζ polypeptide, e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 325), and (ii) a co-stimulatory signaling region comprising a CD28 polypeptide (e.g., an intracellular domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308). In certain embodiments, the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329. In certain embodiments, the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278). In certain embodiments, the VH and VL are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00575] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VH- VL. In certain embodiments, the CAR is designed as “2_28z1XX (vH-vK)” or “ATA018 VH-VK CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 337, which is provided below. MGWSCIIFFLVATATGVHSQVQLQQSGPEVVRPGVSVKISCKGSGYTFTDYAIHWVKQSHAKSLEWI GVISTYNGNTNNNQKFKGKATMTVDKSSSTAYLELARLTSEDSAIYYCARSGTTYWGQGTTLTVSSG GGGSGGGGSGGGGSDVVMTQTPLTLSVTIGQPASISCKSSQSLLDSEGKTYLNWLLQRPGQSPKRLI YLVSKVDSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPHTFGGGTKLELKRAAAIEVM YPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRS NAI-1540294631v3 164 RLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREE YDVLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLSTATK DTFDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 337] [00576] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 337 is set forth in SEQ ID NO: 338, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCCAGGTCCAGC TGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGG CTACACATTCACTGATTATGCTATACACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATT GGAGTTATTAGTACTTACAATGGTAATACAAACAACAACCAGAAGTTTAAGGGCAAGGCCACAATGA CTGTAGACAAATCCTCCAGCACAGCCTATCTGGAACTTGCCAGATTGACATCTGAGGATTCTGCCAT CTATTACTGTGCAAGATCGGGGACGACCTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCAGGT GGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACCCAGACTCCAC TCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAAGTCAGAGCCTCCTAGA TAGTGAAGGAAAGACATATTTGAATTGGTTGTTACAGAGGCCAGGCCAGTCTCCAAAGCGCCTAATC TATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGGACAGATT TCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTGCTGGCAAGGTACACA TTTTCCTCACACGTTCGGTGGTGGGACCAAGCTGGAGCTGAAACGGGCGGCCGCAATTGAAGTTATG TATCCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGAAAGGGAAACACC TTTGTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGTGGTTGGTGGAGT CCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGGAGC AGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACC AGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAGCAGGAGCGCAGA CGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAG TACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACC CTCAGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAGTGAGATTGGGAT GAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGTACAGCCACCAAG GACACCTTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAACAAACTTCTCAC TACTCAAACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 338] [00577] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VL- VH. In certain embodiments, the CAR is designed as “2_28z1XX (vK-vH)” or “ATA018 VK-VH CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 339, which is provided below. NAI-1540294631v3 165 MGWSCIIFFLVATATGVHSDVVMTQTPLTLSVTIGQPASISCKSSQSLLDSEGKTYLNWLLQRPGQS PKRLIYLVSKVDSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCWQGTHFPHTFGGGTKLELKGGG GSGGGGSGGGGSQVQLQQSGPEVVRPGVSVKISCKGSGYTFTDYAIHWVKQSHAKSLEWIGVISTYN GNTNNNQKFKGKATMTVDKSSSTAYLELARLTSEDSAIYYCARSGTTYWGQGTTLTVSSRAAAIEVM YPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRS RLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREE YDVLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLSTATK DTFDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 339] [00578] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 339 is set forth in SEQ ID NO: 340, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCGATGTTGTGA TGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAAG TCAGAGCCTCCTAGATAGTGAAGGAAAGACATATTTGAATTGGTTGTTACAGAGGCCAGGCCAGTCT CCAAAGCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTCAGTGGCAGTG GATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAGGATTTGGGAGTTTATTATTG CTGGCAAGGTACACATTTTCCTCACACGTTCGGTGGTGGGACCAAGCTGGAGCTGAAAGGTGGAGGT GGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAGCTGCAGCAGTCTGGGCCTGAGG TGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACACATTCACTGATTATGC TATACACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATTAGTACTTACAAT GGTAATACAAACAACAACCAGAAGTTTAAGGGCAAGGCCACAATGACTGTAGACAAATCCTCCAGCA CAGCCTATCTGGAACTTGCCAGATTGACATCTGAGGATTCTGCCATCTATTACTGTGCAAGATCGGG GACGACCTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCACGGGCGGCCGCAATTGAAGTTATG TATCCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGAAAGGGAAACACC TTTGTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGTGGTTGGTGGAGT CCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGGAGC AGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACC AGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAGCAGGAGCGCAGA CGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAG TACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACC CTCAGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAGTGAGATTGGGAT GAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGTACAGCCACCAAG GACACCTTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAACAAACTTCTCAC TACTCAAACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 340] NAI-1540294631v3 166 [00579] In certain embodiments, the CAR comprises (a) an extracellular antigen-binding domain comprising (i) a VH that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 25, and (ii) a VL that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 46, (b) a hinge domain comprising a CD28 polypeptide (e.g., a hinge domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308); (c) a transmembrane domain comprising a CD28 polypeptide (e.g., a transmembrane domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308), and (d) an intracellular signaling domain comprising (i) a CD3ζ polypeptide (e.g., a modified human CD3ζ polypeptide, e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 325), and (ii) a co-stimulatory signaling region comprising a CD28 polypeptide (e.g., an intracellular domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308). In certain embodiments, the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329. In certain embodiments, the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278). In certain embodiments, the VH and VL are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00580] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VH- VL. In certain embodiments, the CAR is designed as “4_28z1XX (vH-vK)” or “ATA019 VH-VK CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 341, which is provided below. MGWSCIIFFLVATATGVHSQVQLQQSGPEVVRPGVSVKISCKGSGYTFTDYAMHWVKQSHAKSLEWI GVISTYNGNTNYNQKFKGKATMTVDKSSSTAYMELARLTSEDSAIYYCLKNDYWGQGTSVTVSSGGG GSGGGGSGGGGSDVVMTQTPLTLSVTIGQPASISCKSSQSLLNSDGKTYLNWLLQRPGQSPKRLIYL VSKVDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVFYCWQGTHFPHTFGGGTKLEIKRAAAIEVMYP PPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRL LHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYD VLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLSTATKDT FDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 341] NAI-1540294631v3 167 [00581] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 341 is set forth in SEQ ID NO: 342, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCCAGGTCCAAC TGCAGCAGTCTGGGCCTGAGGTGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGG CTACACATTCACTGATTATGCTATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATT GGAGTTATCAGTACTTACAATGGTAATACAAACTACAACCAGAAGTTTAAGGGCAAGGCCACAATGA CTGTAGACAAATCCTCCAGCACAGCCTATATGGAACTTGCCAGATTGACATCTGAGGATTCTGCCAT CTATTACTGTCTAAAGAACGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGGTGGAGGT GGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGATGTTGTGATGACCCAGACTCCACTCACTT TGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAAGTCAGAGCCTCTTAAATAGTGA TGGAAAGACATATTTGAATTGGTTGTTACAGAGGCCAGGCCAGTCTCCAAAGCGCCTAATCTATCTG GTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTCACTGGCAGTGGATCAGGGACAGATTTCACAC TGAAAATCAGCAGAGTGGAGGCTGAAGATTTGGGAGTTTTTTATTGCTGGCAAGGTACACATTTTCC TCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAACGGGCGGCCGCAATTGAAGTTATGTATCCT CCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGAAAGGGAAACACCTTTGTC CAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGC TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGGAGCAGGCTC CTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCT ATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCC CGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGAT GTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGG AAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAGTGAGATTGGGATGAAAGG CGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGTACAGCCACCAAGGACACC TTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAACAAACTTCTCACTACTCA AACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 342] [00582] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VL- VH. In certain embodiments, the CAR is designed as “4_28z1XX (vK-vH)” or “ATA019 VK-VH CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 343, which is provided below. MGWSCIIFFLVATATGVHSDVVMTQTPLTLSVTIGQPASISCKSSQSLLNSDGKTYLNWLLQRPGQS PKRLIYLVSKVDSGVPDRFTGSGSGTDFTLKISRVEAEDLGVFYCWQGTHFPHTFGGGTKLEIKGGG GSGGGGSGGGGSQVQLQQSGPEVVRPGVSVKISCKGSGYTFTDYAMHWVKQSHAKSLEWIGVISTYN GNTNYNQKFKGKATMTVDKSSSTAYMELARLTSEDSAIYYCLKNDYWGQGTSVTVSSRAAAIEVMYP NAI-1540294631v3 168 PPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRL LHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYD VLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLSTATKDT FDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 343] [00583] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 343 is set forth in SEQ ID NO: 344, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCGATGTTGTGA TGACCCAGACTCCACTCACTTTGTCGGTTACCATTGGACAACCAGCCTCCATCTCTTGCAAGTCAAG TCAGAGCCTCTTAAATAGTGATGGAAAGACATATTTGAATTGGTTGTTACAGAGGCCAGGCCAGTCT CCAAAGCGCCTAATCTATCTGGTGTCTAAAGTGGACTCTGGAGTCCCTGACAGGTTCACTGGCAGTG GATCAGGGACAGATTTCACACTGAAAATCAGCAGAGTGGAGGCTGAAGATTTGGGAGTTTTTTATTG CTGGCAAGGTACACATTTTCCTCACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAAGGTGGAGGT GGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTCAGGTCCAACTGCAGCAGTCTGGGCCTGAGG TGGTGAGGCCTGGGGTCTCAGTGAAGATTTCCTGCAAGGGTTCCGGCTACACATTCACTGATTATGC TATGCACTGGGTGAAGCAGAGTCATGCAAAGAGTCTAGAGTGGATTGGAGTTATCAGTACTTACAAT GGTAATACAAACTACAACCAGAAGTTTAAGGGCAAGGCCACAATGACTGTAGACAAATCCTCCAGCA CAGCCTATATGGAACTTGCCAGATTGACATCTGAGGATTCTGCCATCTATTACTGTCTAAAGAACGA CTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCACGGGCGGCCGCAATTGAAGTTATGTATCCT CCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGAAAGGGAAACACCTTTGTC CAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCTGGC TTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGGAGCAGGCTC CTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGCCCT ATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAGCAGGAGCGCAGACGCCCC CGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGAT GTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGG AAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAGTGAGATTGGGATGAAAGG CGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGTACAGCCACCAAGGACACC TTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAACAAACTTCTCACTACTCA AACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 344] [00584] In certain embodiments, the CAR comprises (a) an extracellular antigen-binding domain comprising (i) a VH that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 133, and (ii) a VL that NAI-1540294631v3 169 comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 136, (b) a hinge domain comprising a CD28 polypeptide (e.g., a hinge domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308); (c) a transmembrane domain comprising a CD28 polypeptide (e.g., a transmembrane domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308), and (d) an intracellular signaling domain comprising (i) a CD3ζ polypeptide (e.g., a modified human CD3ζ polypeptide, e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 325), and (ii) a co-stimulatory signaling region comprising a CD28 polypeptide (e.g., an intracellular domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308). In certain embodiments, the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329. In certain embodiments, the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278). In certain embodiments, the VH and VL are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00585] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VH- VL. In certain embodiments, the CAR is designed as “8_28z1XX (vH-vK)” or “ATA020 VH-VK CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 345, which is provided below. MGWSCIIFFLVATATGVHSQVQLKQSGPGLVQPSQSLSITCTVSGFSLNSYGVHWVRQSPGKGLEWL GAIWSSGSTDYNAPFISRLSISKDNSKNQVFFKMNSLQVDDTAIYFCAGNPESDHYYGYEAMDSWGQ GTSVTVSSGGGGSGGGGSGGGGSDIQMTQSPASLSASVGETVTITCRASGNIHNYLAWYQQRQGKSP QLLVYNAKTLADGVPSRFSGSGSGTQYSLKIISLQPEDFGSYYCQHFWTTPFTFGSGTKLEIKRAAA IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVR SKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLG RREEYDVLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLS TATKDTFDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 345] [00586] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 345 is set forth in SEQ ID NO: 346, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCCAGGTGCAGC TGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGG NAI-1540294631v3 170 TTTCTCATTAAATAGCTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTG GGAGCGATATGGAGTAGTGGAAGCACAGACTATAATGCACCTTTCATATCCAGACTGAGCATCAGCA AGGACAACTCCAAGAACCAAGTTTTCTTTAAAATGAACAGTCTGCAAGTTGATGACACAGCCATATA TTTTTGTGCCGGAAATCCAGAATCGGATCATTACTACGGCTACGAGGCCATGGACTCCTGGGGTCAA GGAACCTCAGTCACCGTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGAT CTGACATCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCAC ATGTCGAGCAAGTGGGAATATTCACAATTATTTAGCATGGTATCAGCAGAGACAGGGAAAATCTCCT CAGCTCCTGGTCTATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGTGGCAGTGGAT CAGGAACACAATATTCTCTCAAGATCATCAGCCTTCAGCCTGAAGATTTTGGGAGTTATTACTGTCA ACATTTTTGGACTACTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAACGGGCGGCCGCA ATTGAAGTTATGTATCCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGA AAGGGAAACACCTTTGTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGT GGTTGGTGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGG AGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCC GCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAG CAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGA CGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGA GAAGGAAGAACCCTCAGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAG TGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGT ACAGCCACCAAGGACACCTTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAA CAAACTTCTCACTACTCAAACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 346] [00587] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VL- VH. In certain embodiments, the CAR is designed as “8_28z1XX (vK-vH)” or “ATA020 VK-VH CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 347, which is provided below. MGWSCIIFFLVATATGVHSDIQMTQSPASLSASVGETVTITCRASGNIHNYLAWYQQRQGKSPQLLV YNAKTLADGVPSRFSGSGSGTQYSLKIISLQPEDFGSYYCQHFWTTPFTFGSGTKLEIKGGGGSGGG GSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLNSYGVHWVRQSPGKGLEWLGAIWSSGSTDYN APFISRLSISKDNSKNQVFFKMNSLQVDDTAIYFCAGNPESDHYYGYEAMDSWGQGTSVTVSSRAAA IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVR SKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLG NAI-1540294631v3 171 RREEYDVLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLS TATKDTFDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 347] [00588] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 347 is set forth in SEQ ID NO: 348, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCGACATCCAGA TGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACATGTCGAGCAAG TGGGAATATTCACAATTATTTAGCATGGTATCAGCAGAGACAGGGAAAATCTCCTCAGCTCCTGGTC TATAATGCAAAAACCTTAGCCGATGGTGTGCCATCAAGGTTCAGTGGCAGTGGATCAGGAACACAAT ATTCTCTCAAGATCATCAGCCTTCAGCCTGAAGATTTTGGGAGTTATTACTGTCAACATTTTTGGAC TACTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAAGGTGGAGGTGGATCAGGTGGAGGT GGATCTGGTGGAGGTGGATCTCAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCAC AGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAAATAGCTATGGTGTACACTGGGTTCG CCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGCGATATGGAGTAGTGGAAGCACAGACTATAAT GCACCTTTCATATCCAGACTGAGCATCAGCAAGGACAACTCCAAGAACCAAGTTTTCTTTAAAATGA ACAGTCTGCAAGTTGATGACACAGCCATATATTTTTGTGCCGGAAATCCAGAATCGGATCATTACTA CGGCTACGAGGCCATGGACTCCTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCACGGGCGGCCGCA ATTGAAGTTATGTATCCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGA AAGGGAAACACCTTTGTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGT GGTTGGTGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGG AGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCC GCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAG CAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGA CGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGA GAAGGAAGAACCCTCAGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAG TGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGT ACAGCCACCAAGGACACCTTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAA CAAACTTCTCACTACTCAAACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 348] [00589] In certain embodiments, the CAR comprises (a) an extracellular antigen-binding domain comprising (i) a VH that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 151, and (ii) a VL that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 NAI-1540294631v3 172 consisting of the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 154, (b) a hinge domain comprising a CD28 polypeptide (e.g., a hinge domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308); (c) a transmembrane domain comprising a CD28 polypeptide (e.g., a transmembrane domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308), and (d) an intracellular signaling domain comprising (i) a CD3ζ polypeptide (e.g., a modified human CD3ζ polypeptide, e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 325), and (ii) a co-stimulatory signaling region comprising a CD28 polypeptide (e.g., an intracellular domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308). In certain embodiments, the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329. In certain embodiments, the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278). In certain embodiments, the VH and VL are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00590] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VH- VL. In certain embodiments, the CAR is designed as “9_28z1XX (vH-vK)” or “ATA021 VH-VK CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 349, which is provided below. MGWSCIIFFLVATATGVHSEVQLQQSGAEHVKPGTSVRLSCTASGFNIKDAYIHWVKQRPEQGLEWI GGIDPENDNTEYDPKFPGKATVVADTSSNTAYLQLSSLTSEDTAVYYCARRPLGLPFDYWGQGTTLT VSSGGGGSGGGGSGGGGSDIVLSQSPSSLAVSVGENVTMNCKSSQRLLDSSNQRNYLAWYQQKPGQS PQLLIFWASTRESGVPDRFTGSGSGTDFTLTIRSVKAEDLAVYYCQQYYSYPFTFGSGTKLEIKRAA AIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWV RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNL GRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGL STATKDTFDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 349] [00591] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 349 is set forth in SEQ ID NO: 350, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCGAGGTTCAGC TTCAGCAGTCTGGGGCAGAGCATGTGAAGCCAGGGACCTCAGTCAGGTTGTCCTGCACAGCTTCTGG CTTTAACATTAAAGACGCCTATATACACTGGGTGAAGCAGAGGCCTGAACAGGGCCTGGAGTGGATT NAI-1540294631v3 173 GGAGGGATTGATCCTGAGAATGACAATACTGAATATGACCCTAAATTCCCCGGCAAGGCCACTGTTG TGGCCGACACATCCTCCAACACAGCCTACCTGCAACTCAGCAGCCTGACATCTGAGGACACTGCCGT CTATTACTGTGCTAGAAGACCTCTCGGACTACCTTTTGACTACTGGGGCCAAGGCACCACTCTCACA GTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGACATTGTGCTGT CACAGTCTCCATCCTCCCTAGCTGTGTCAGTTGGAGAGAACGTTACTATGAACTGCAAGTCCAGTCA GAGACTTTTAGATAGTAGCAATCAAAGGAACTATTTGGCCTGGTACCAGCAGAAACCAGGGCAGTCT CCTCAACTGCTGATTTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTG GATCTGGGACAGATTTCACTCTCACCATCAGAAGTGTGAAGGCTGAAGACCTGGCAGTTTATTACTG TCAGCAATATTATAGCTATCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAACGGGCGGCC GCAATTGAAGTTATGTATCCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATG TGAAAGGGAAACACCTTTGTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGT GGTGGTTGGTGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTG AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCA CCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTT CAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTA GGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGC CGAGAAGGAAGAACCCTCAGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTT CAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTC AGTACAGCCACCAAGGACACCTTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAG CAACAAACTTCTCACTACTCAAACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 350] [00592] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VL- VH. In certain embodiments, the CAR is designed as “9_28z1XX (vK-vH)” or “ATA021 VK-VH CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 351, which is provided below. MGWSCIIFFLVATATGVHSDIVLSQSPSSLAVSVGENVTMNCKSSQRLLDSSNQRNYLAWYQQKPGQ SPQLLIFWASTRESGVPDRFTGSGSGTDFTLTIRSVKAEDLAVYYCQQYYSYPFTFGSGTKLEIKGG GGSGGGGSGGGGSEVQLQQSGAEHVKPGTSVRLSCTASGFNIKDAYIHWVKQRPEQGLEWIGGIDPE NDNTEYDPKFPGKATVVADTSSNTAYLQLSSLTSEDTAVYYCARRPLGLPFDYWGQGTTLTVSSRAA AIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWV RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNL GRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGL STATKDTFDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 351] NAI-1540294631v3 174 [00593] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 351 is set forth in SEQ ID NO: 352, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCGACATTGTGC TGTCACAGTCTCCATCCTCCCTAGCTGTGTCAGTTGGAGAGAACGTTACTATGAACTGCAAGTCCAG TCAGAGACTTTTAGATAGTAGCAATCAAAGGAACTATTTGGCCTGGTACCAGCAGAAACCAGGGCAG TCTCCTCAACTGCTGATTTTCTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCA GTGGATCTGGGACAGATTTCACTCTCACCATCAGAAGTGTGAAGGCTGAAGACCTGGCAGTTTATTA CTGTCAGCAATATTATAGCTATCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAAGGTGGA GGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGAGGTTCAGCTTCAGCAGTCTGGGGCAG AGCATGTGAAGCCAGGGACCTCAGTCAGGTTGTCCTGCACAGCTTCTGGCTTTAACATTAAAGACGC CTATATACACTGGGTGAAGCAGAGGCCTGAACAGGGCCTGGAGTGGATTGGAGGGATTGATCCTGAG AATGACAATACTGAATATGACCCTAAATTCCCCGGCAAGGCCACTGTTGTGGCCGACACATCCTCCA ACACAGCCTACCTGCAACTCAGCAGCCTGACATCTGAGGACACTGCCGTCTATTACTGTGCTAGAAG ACCTCTCGGACTACCTTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCACGGGCGGCC GCAATTGAAGTTATGTATCCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATG TGAAAGGGAAACACCTTTGTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGT GGTGGTTGGTGGAGTCCTGGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTG AGGAGTAAGAGGAGCAGGCTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCA CCCGCAAGCATTACCAGCCCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTT CAGCAGGAGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTA GGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGC CGAGAAGGAAGAACCCTCAGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTT CAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTC AGTACAGCCACCAAGGACACCTTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAG CAACAAACTTCTCACTACTCAAACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 352] [00594] In certain embodiments, the CAR comprises (a) an extracellular antigen-binding domain comprising (i) a VH that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 260, and (ii) a VL that comprises a CDR1 consisting of the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 consisting of the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 consisting of the amino acid sequence set forth in SEQ ID NO: 261, (b) a hinge domain comprising a CD28 NAI-1540294631v3 175 polypeptide (e.g., a hinge domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 114 to 153 of SEQ ID NO: 308); (c) a transmembrane domain comprising a CD28 polypeptide (e.g., a transmembrane domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 154 to 179 of SEQ ID NO: 308), and (d) an intracellular signaling domain comprising (i) a CD3ζ polypeptide (e.g., a modified human CD3ζ polypeptide, e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 325), and (ii) a co-stimulatory signaling region comprising a CD28 polypeptide (e.g., an intracellular domain of human CD28 or a portion thereof, e.g., a CD28 polypeptide consisting of the amino acid sequence of amino acids 180 to 220 of SEQ ID NO: 308). In certain embodiments, the CAR further comprises a self-cleaving P2A peptide (e.g., one consisting of the amino acid sequence set forth in SEQ ID NO: 329. In certain embodiments, the CAR further comprises a signal peptide (e.g., one consisting of amino acid sequence set forth in SEQ ID NO: 278). In certain embodiments, the VH and VL are linked via a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1. [00595] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VH- VL. In certain embodiments, the CAR is designed as “16_28z1XX (vH-vK)” or “ATA022 VH-VK CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 353, which is provided below. MGWSCIIFFLVATATGVHSQVQLQQPGSELVRPGASVKLSCKASGYTFTNYWLHWVKQRPGQGLEWI GNIYPGYGTSNYDEKFTNKAALTIDTSSSTAYMQLSSLTSEDSAVYFCTRGGYDYYFDYWGQGTTLT VSSGGGGSGGGGSGGGGSDIQMNQSPSSLSASLGDTITITCHASQNINVWLSWYQQTPGNIPKLLIY KASNLHTGVPSRFSGSGSGTGFTLTISSLQPEDIATYYCQQGQSYPWTFGGGTKLEIKRAAAIEVMY PPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSR LLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEY DVLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLSTATKD TFDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 353] [00596] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 353 is set forth in SEQ ID NO: 354, which is provided below. ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCCAGGTCCAAC TGCAGCAACCTGGGTCTGAGCTGGTGAGGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGG CTACACATTCACCAACTACTGGTTGCACTGGGTGAAGCAAAGGCCTGGACAAGGCCTTGAGTGGATT GGAAATATTTATCCTGGTTATGGTACTTCTAACTACGATGAGAAGTTCACGAACAAGGCCGCACTGA CTATAGACACATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGT NAI-1540294631v3 176 CTATTTCTGTACAAGAGGGGGATATGATTACTACTTTGACTACTGGGGCCAAGGCACCACTCTCACA GTCTCCTCAGGTGGAGGTGGATCAGGTGGAGGTGGATCTGGTGGAGGTGGATCTGACATTCAGATGA ACCAGTCTCCATCCAGTCTGTCTGCATCCCTTGGAGACACAATTACCATCACTTGCCATGCCAGTCA GAACATTAATGTTTGGTTAAGCTGGTACCAGCAGACACCAGGAAATATTCCTAAACTATTGATCTAT AAGGCTTCCAACTTGCACACAGGCGTCCCATCAAGGTTTAGTGGCAGTGGATCTGGAACAGGTTTCA CATTAACCATCAGCAGTCTGCAGCCTGAAGACATTGCCACTTACTACTGTCAACAGGGTCAAAGTTA TCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACGGGCGGCCGCAATTGAAGTTATGTAT CCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGAAAGGGAAACACCTTT GTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCT GGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGGAGCAGG CTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGC CCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAGCAGGAGCGCAGACGC CCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTAC GATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTC AGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAGTGAGATTGGGATGAA AGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGTACAGCCACCAAGGAC ACCTTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAACAAACTTCTCACTAC TCAAACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 354] [00597] In certain embodiments, the VH and VL are positioned from the N- to the C-terminus: VL- VH. In certain embodiments, the CAR is designed as “16_28z1XX (vK-vH)” or “ATA022 VK-VH CAR”. In certain embodiments, the CAR comprises the amino acid sequence set forth in SEQ ID NO: 355, which is provided below. MGWSCIIFFLVATATGVHSDIQMNQSPSSLSASLGDTITITCHASQNINVWLSWYQQTPGNIPKLLI YKASNLHTGVPSRFSGSGSGTGFTLTISSLQPEDIATYYCQQGQSYPWTFGGGTKLEIKGGGGSGGG GSGGGGSQVQLQQPGSELVRPGASVKLSCKASGYTFTNYWLHWVKQRPGQGLEWIGNIYPGYGTSNY DEKFTNKAALTIDTSSSTAYMQLSSLTSEDSAVYFCTRGGYDYYFDYWGQGTTLTVSSRAAAIEVMY PPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSR LLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEY DVLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLSTATKD TFDALHMQALPPRGSGATNFSLLKQAGDVEENPGP [SEQ ID NO: 355] [00598] An exemplary nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 355 is set forth in SEQ ID NO: 356, which is provided below. NAI-1540294631v3 177 ATGGGTTGGAGCTGTATCATCTTCTTTCTGGTAGCAACAGCTACAGGTGTGCACTCCGACATTCAGA TGAACCAGTCTCCATCCAGTCTGTCTGCATCCCTTGGAGACACAATTACCATCACTTGCCATGCCAG TCAGAACATTAATGTTTGGTTAAGCTGGTACCAGCAGACACCAGGAAATATTCCTAAACTATTGATC TATAAGGCTTCCAACTTGCACACAGGCGTCCCATCAAGGTTTAGTGGCAGTGGATCTGGAACAGGTT TCACATTAACCATCAGCAGTCTGCAGCCTGAAGACATTGCCACTTACTACTGTCAACAGGGTCAAAG TTATCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAAGGTGGAGGTGGATCAGGTGGAGGT GGATCTGGTGGAGGTGGATCTCAGGTCCAACTGCAGCAACCTGGGTCTGAGCTGGTGAGGCCTGGAG CTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACATTCACCAACTACTGGTTGCACTGGGTGAA GCAAAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTTATCCTGGTTATGGTACTTCTAACTAC GATGAGAAGTTCACGAACAAGGCCGCACTGACTATAGACACATCCTCCAGCACAGCCTACATGCAGC TCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTTCTGTACAAGAGGGGGATATGATTACTACTT TGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCACGGGCGGCCGCAATTGAAGTTATGTAT CCTCCTCCTTACCTAGACAATGAGAAGAGCAATGGAACCATTATCCATGTGAAAGGGAAACACCTTT GTCCAAGTCCCCTATTTCCCGGACCTTCTAAGCCCTTTTGGGTGCTGGTGGTGGTTGGTGGAGTCCT GGCTTGCTATAGCTTGCTAGTAACAGTGGCCTTTATTATTTTCTGGGTGAGGAGTAAGAGGAGCAGG CTCCTGCACAGTGACTACATGAACATGACTCCCCGCCGCCCCGGGCCCACCCGCAAGCATTACCAGC CCTATGCCCCACCACGCGACTTCGCAGCCTATCGCTCCAGAGTGAAGTTCAGCAGGAGCGCAGACGC CCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTAC GATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTC AGGAAGGCCTGTTCAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTTCAGTGAGATTGGGATGAA AGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTTCCAGGGGCTCAGTACAGCCACCAAGGAC ACCTTCGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCGGATCTGGAGCAACAAACTTCTCACTAC TCAAACAAGCAGGGGACGTGGAGGAGAATCCCGGACCC [SEQ ID NO: 356] 5.4. Cells [00599] The presently disclosed subject matter provides cells comprising a presently disclosed GPC3-targeted CAR (e.g., one disclosed in Section 5.3). In certain embodiments, the cell is selected from the group consisting of cells of lymphoid lineage and cells of myeloid lineage. In certain embodiments, the cell is an immunoresponsive cell. In certain embodiments, the immunoresponsive cell is a cell of lymphoid lineage. [00600] In certain embodiments, the cell is a cell of the lymphoid lineage. Cells of the lymphoid lineage can provide production of antibodies, regulation of cellular immune system, detection of foreign agents in the blood, detection of cells foreign to the host, and the like. Non-limiting examples of cells of the lymphoid lineage include T cells, Natural Killer (NK) cells, B cells, NAI-1540294631v3 178 dendritic cells, and stem cells from which lymphoid cells may be differentiated. In certain embodiments, the stem cell is a pluripotent stem cell (e.g., embryonic stem cell or an induced pluripotent stem cell). [00601] In certain embodiments, the cell is an immune effector cell. Immune effector cells can be obtained from a number of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. Immune effector cells can be obtained from blood collected from a subject using any number of techniques known to the skilled artisan, such as Ficoll™ separation. For example, cells from the circulating blood of an individual may be obtained by apheresis. In certain embodiments,, immune effector cells are isolated from peripheral blood lymphocytes by lysing the red blood cells and depleting the monocytes, for example, by centrifugation through a PERCOLL™ gradient or by counterflow centrifugal elutriation. A specific subpopulation of immune effector cells can be further isolated by positive or negative selection techniques. For example, immune effector cells can be isolated using a combination of antibodies directed to surface markers unique to the positively selected cells, e.g., by incubation with antibody-conjugated beads for a time period sufficient for positive selection of the desired immune effector cells. Alternatively, enrichment of immune effector cells population can be accomplished by negative selection using a combination of antibodies directed to surface markers unique to the negatively selected cells. [00602] In certain embodiments, the immune effector cells comprise any leukocyte involved in defending the body against infectious disease and foreign materials. For example, the immune effector cells can comprise lymphocytes, monocytes, macrophages, dendritic cells, mast cells, neutrophils, basophils, eosinophils, or any combinations thereof. For example, the immune effector cells can comprise T lymphocytes, preferably cytotoxic T lymphocytes (CTLs). [00603] In certain embodiments, the cell is a T cell. T cells can be lymphocytes that mature in the thymus and are chiefly responsible for cell-mediated immunity. T cells are involved in the adaptive immune system. T cells or T lymphocytes can be distinguished from other lymphocytes, such as B cells and natural killer cells (NK cells), by the presence of a T-cell receptor (TCR) on the cell surface. They are called T cells because they mature in the thymus (although some also mature in the tonsils). There are several subsets of T cells, each with a distinct function. [00604] The T cells of the presently disclosed subject matter can be any type of T cells, including, but not limited to, helper T cells, cytotoxic T cells, memory T cells (including central memory T cells, stem-cell-like memory T cells (or stem-like memory T cells), and two types of effector memory T cells: e.g., TEM cells and TEMRA cells), Regulatory T cells (also known as suppressor T NAI-1540294631v3 179 cells or Treg), tumor-infiltrating lymphocyte (TIL), Natural killer T cells (NK-T cells), mucosal associated invariant T cells, αβ T cells, and γδ T cells. [00605] In certain embodiments, the cell comprising the presently disclosed GPC3-targeted CAR is a helper T cell or CD4+ T cell. Helper T cells (or T helper cells, or TH cells) assist other white blood cells in immunologic processes, including maturation of B cells into plasma cells and memory B cells, and activation of cytotoxic T cells and macrophages. These cells are also known as CD4+ T cells because they express the CD4 glycoprotein on their surface. Helper T cells become activated when they are presented with peptide antigens by MHC class II molecules, which are expressed on the surface of antigen-presenting cells (APCs). Once activated, they divide rapidly and secrete small proteins called cytokines that regulate or assist in the active immune response. These cells can differentiate into one of several subtypes, including TH1, TH2, TH3, TH17, TH9, or TFH, which secrete different cytokines to facilitate a different type of immune response. [00606] In certain embodiments, the cell comprising the presently disclosed GPC3-targeted CAR is a cytotoxic T cell or CD8+ T cell. Cytotoxic T cells (TC cells, or CTLs) destroy virally infected cells and tumor cells and are also implicated in transplant rejection. These cells are also known as CD8+ T cells since they express the CD8 glycoprotein at their surface. These cells recognize their targets by binding to antigen associated with MHC class I molecules, which are present on the surface of all nucleated cells. Through IL-10, adenosine and other molecules secreted by regulatory T cells, the CD8+ cells can be inactivated to an anergic state, which prevents autoimmune diseases. [00607] In certain embodiments, the cell comprising the presently disclosed GPC3-targeted CAR is a memory T cell. Memory T cells are a subset of antigen-specific T cells that persist long-term after an infection has resolved. They quickly expand to large numbers of effector T cells upon re- exposure to their cognate antigen, thus providing the immune system with “memory” against past infections. Memory cells may be either CD4+ or CD8+. Memory T cells typically express the cell surface protein CD45RO. [00608] In certain embodiments, the cell comprising the presently disclosed GPC3-targeted CAR is a regulatory T cell. Regulatory T cells (Treg cells), formerly known as suppressor T cells, are crucial for the maintenance of immunological tolerance. Their major role is to shut down T cell- mediated immunity toward the end of an immune reaction and to suppress auto-reactive T cells that escaped the process of negative selection in the thymus. Two major classes of CD4+ Treg cells have been described — naturally occurring Treg cells and adaptive Treg cells. [00609] In certain embodiments, the cell comprising the presently disclosed GPC3-targeted CAR is a Natural killer T cell. Natural killer T (NKT) cells (not to be confused with natural killer (NK) NAI-1540294631v3 180 cells) bridge the adaptive immune system with the innate immune system. Unlike conventional T cells that recognize peptide antigens presented by major histocompatibility complex (MHC) molecules, NKT cells recognize glycolipid antigen presented by a molecule called CD1d. [00610] In certain embodiments, the cell comprising the presently disclosed GPC3-targeted CAR is a γδ T cell. γδ T cells possess a distinct T-cell receptor (TCR) having one γ chain and one δ chain instead of α and β chains. [00611] In certain embodiments, the cell comprising the presently disclosed GPC3-targeted CAR is an innate lymphoid cell (ILC), a cytokine induced killer (CIK) cell, or a lymphokine activated killer (LAK) cell. [00612] In certain embodiments, the cell is a T cell, and the presently disclosed GPC3-targeted CAR is integrated at a locus within the genome of the T cell. Non-limiting examples of the loci include a TRAC locus, a TRBC locus, a TRDC locus, and a TRGC locus. In certain embodiments, the locus is a TRAC locus or a TRBC locus. Methods of targeting a CAR to a site within the genome of T cell are disclosed in WO2017180989 and Eyquem et al., Nature. (2017 Mar 2); 543(7643): 113-117, both of which are incorporated by reference in their entireties. [00613] In certain embodiments, the T cells comprise a mixture of CD4+ T cells and CD8+ T cells. In certain embodiments, the T cells are enriched for one or more subsets based on cell surface marker expression profile. [00614] In certain embodiments, the cell is a Natural killer cell. Natural killer (NK) cells can be lymphocytes that are part of cell-mediated immunity and act during the innate immune response. NK cells do not require prior activation in order to perform their cytotoxic effect on target cells. Unlike cytotoxic CD8+ T cells, NK cells launch cytotoxicity against tumor cells without the requirement for prior sensitization and can also eradicate MHC-I-negative cells. In certain embodiments, the cell is a genetically modified NK cell. In certain embodiments, the cell is an edited NK cell. In certain embodiments, the cell is a NK cell derived from a stem cell. In certain embodiments, the cell is a NK cell derived from a pluripotent stem cell. In certain embodiments, the cell is an induced pluripotent stem cell (iPSC)-derived NK cell. [00615] The presently disclosed cells (e.g., T cells or NK cells) can be autologous, non- autologous (e.g., allogeneic), or derived in vitro from engineered progenitor or stem cells. In certain embodiments, the cells comprising a presently disclosed GPC3-targeted CAR are obtained from the subject to be treated (i.e. are autologous). In certain embodiments, the cells comprising a presently disclosed GPC3-targeted CAR are allogeneic to the subject to be treated. NAI-1540294631v3 181 [00616] The cells of the presently disclosed subject matter can be cells of the myeloid lineage. Non-limiting examples of cells of the myeloid lineage include monocytes, macrophages, neutrophils, dendritic cells, basophils, neutrophils, eosinophils, megakaryocytes, mast cell, erythrocyte, thrombocytes, and stem cells from which myeloid cells may be differentiated. In certain embodiments, the stem cell is a pluripotent stem cell (e.g., an embryonic stem cell or an induced pluripotent stem cell). [00617] Epstein-Barr virus (EBV)-induced lymphoproliferative diseases (EBV-LPDs) and other EBV-associated cancers are a significant cause of morbidity and mortality for recipients of allogeneic hematopoietic cell transplantation (HCT) or solid organ transplants (SOT), particularly in those who have received certain T-cell reactive Abs to prevent or treat GVHD. Prophylaxis and treatment by the adoptive transfer of autologous or allogeneic EBV-specific cytotoxic T cells and the subsequent long-term restoration of immunity against EBV-associated lymphoproliferation have provided positive outcomes in the management of these uniformly fatal complications of allogeneic tissue transfer. Therefore, in some embodiments, the disclosed immune effector cells that comprise one or more of the CAR polypeptides of the present invention are allogeneic or autologous EBV- specific cytotoxic T lymphocytes (CTLs). For example, generation of EBV-specific cytotoxic T cells may involve isolating PBMCs from of an EBV-seropositive autologous or allogenic donor and enriching them for T cells by depletion of monocytes and NK cells. EBV-specific cytotoxic T cells may also be produced by contacting donor PBMCs or purified donor T cells with a “stimulator” cell that expresses one or more EBV antigen(s) and presents the EBV antigen(s) to unstimulated T cells, thereby causing stimulation and expansion of EBV-specific CTLs. EBV antigens include, for example, latent membrane protein (LMP) and EBV nuclear antigen (EBNA) proteins, such as LMP- 1, LMP-2A, and LMP-2B and EBNA-1, EBNA-2, EBNA-3A, EBNA-3B, EBNA-3C and EBNA- LP. Cytotoxic T cells that comprise T cell receptor(s) which recognize one or more EBV-specific antigens are deemed to have been “sensitized” to those EBV antigen(s) and are therefore termed “EBV-sensitized cytotoxic T cells” herein. Known methods for generating allogeneic or autologous EBV-specific cytotoxic T cell populations that may comprise one or more of the CAR polypeptides of the present invention are described, for example, in Barker et al., Blood 116(23):5045-49 (2010); Doubrovina et al., Blood 119(11):2644-56 (2012); Koehne et al., Blood 99(5):1730-40 (2002); and Smith et al., Cancer Res.72(5):1116-25 (2012), which are incorporated by reference for these teachings. Similarly, cytotoxic T cells may be “sensitized” to other viral antigens, including cytomegalovirus (CMV), papillomavirus (e.g., HPV), adenovirus, polyomavirus (e.g., BKV, JCV, and Merkel cell virus), retrovirus (e.g., HTLV-I, also including lentivirus such as HIV), picomavirus NAI-1540294631v3 182 (e.g., Hepatitis A virus), hepadnavirus (e.g., Hepatitis B virus), hepacivirus (e.g., Hepatitis C virus), deltavirus (e.g., Hepatitis D virus), hepevirus (e.g., Hepatitis E virus), and the like. In certain embodiments, the T cells used for generating the CAR-T cells of the invention are polyfunctional T- cells, i.e., those T cells that are capable of inducing multiple immune effector functions, that provide a more effective immune response to a pathogen than do cells that produce, for example, only a single immune effector (e.g. a single biomarker such as a cytokine or CD107a). Less-polyfunctional, monofunctional, or even “exhausted” T cells may dominate immune responses during chronic infections, thus negatively impacting protection against virus-associated complications. In certain embodiments, the presently disclosed CAR-T cells are polyfunctional. In certain embodiments, at least 50% of the T cells used for generating the presently disclosed CAR-T cells are CD4+ T cells. In certain embodiments, the T cells are less than 50% CD4+ T cells. In certain embodiments, the T cells are predominantly CD4+ T cells. In certain embodiments, at least 50% of the T cells used for generating the presently disclosed CAR-T cells are CD8+ T cells. In certain embodiments, the T cells are less than 50% CD8+ T cells. In certain embodiments, the T cells are predominantly CD8+ T cells. In some embodiments, the T cells (e.g., the sensitized T cells and/or CAR-T cells described herein) are stored in a cell library or bank before they are administered to the subject. [00618] In certain embodiments, the cells can be transduced with the presently disclosed GPC3- targeted CAR such that the cells express the GPC3-targeted CAR. 5.5. Anti-GPC3 Antibodies [00619] The presently disclosed subject matter provides anti-GPC3 antibodies and antigen- binding fragments thereof. The antibodies can be polyclonal antibodies, monoclonal antibodies, humanized antibodies, human antibodies, multispecific antibodies, bispecific antibodies, or hetero- conjugate antibodies, as well as variants thereof having increased or decreased affinity or other properties. [00620] In certain embodiments, the presently disclosed anti-GPC3 antibody and antigen-binding fragment thereof binds to human GPC3. 5.5.1. Monospecific Antibodies and Antigen-binding Fragments Thereof [00621] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL). [00622] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 29; and/or ii) the VL comprises NAI-1540294631v3 183 a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 30. SEQ ID NO: 29 and SEQ ID NO: 30 are disclosed in Table 1. [00623] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 47; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 48. SEQ ID NO: 47 and SEQ ID NO: 48 are disclosed in Table 2. [00624] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 62; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 63. SEQ ID NO: 62 and SEQ ID NO: 63 are disclosed in Table 3. [00625] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 75; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 76. SEQ ID NO: 75 and SEQ ID NO: 76 are disclosed in Table 4. [00626] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 89; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 90. SEQ ID NO: 89 and SEQ ID NO: 90 are disclosed in Table 5. [00627] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 101; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 102. SEQ ID NO: 101 and SEQ ID NO: 102 are disclosed in Table 6. [00628] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 119; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 120. SEQ ID NO: 119 and SEQ ID NO: 120 are disclosed in Table 7. [00629] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 137; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 138. SEQ ID NO: 137 and SEQ ID NO: 138 are disclosed in Table 8. [00630] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 155; and/or ii) the VL NAI-1540294631v3 184 comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 156. SEQ ID NO: 155 and SEQ ID NO: 156 are disclosed in Table 9. [00631] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 170; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 171. SEQ ID NO: 170 and SEQ ID NO: 171 are disclosed in Table 10. [00632] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 183; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 184. SEQ ID NO: 183 and SEQ ID NO: 184 are disclosed in Table 11. [00633] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 201; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 202. SEQ ID NO: 201 and SEQ ID NO: 202 are disclosed in Table 12. [00634] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 215; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 216. SEQ ID NO: 215 and SEQ ID NO: 216 are disclosed in Table 13. [00635] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 233; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 234. SEQ ID NO: 233 and SEQ ID NO: 234 are disclosed in Table 14. [00636] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 246; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 247. SEQ ID NO: 246 and SEQ ID NO: 247 are disclosed in Table 15. [00637] In certain embodiments, the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 as set forth in a VH comprising the amino acid sequence of SEQ ID NO: 262; and/or ii) the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 as set forth in a VL comprising the amino acid sequence of SEQ ID NO: 263. SEQ ID NO: 262 and SEQ ID NO: 263 are disclosed in Table 16. [00638] The VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 can be identified according to any known CDR numbering systems, which include but are not limited to, Kabat numbering system, Chothia numbering system, AbM numbering system. Contact numbering NAI-1540294631v3 185 system, and IMGT® Information System. In certain embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system. [00639] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28. [00640] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 23, SEQ ID NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, and SEQ ID NO: 28 are disclosed in Table 1. [00641] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification NAI-1540294631v3 186 thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00642] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, and SEQ ID NO: 46 are disclosed in Table 2. [00643] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00644] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and a VL comprising a NAI-1540294631v3 187 CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61 SEQ ID NO: 26, SEQ ID NO: 27, and SEQ ID NO: 46 are disclosed in Table 3. [00645] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00646] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 23, SEQ ID NO: 60, SEQ ID NO: 25, SEQ ID NO: 74, SEQ ID NO: 45, and SEQ ID NO: 46 are disclosed in Table 4. [00647] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set NAI-1540294631v3 188 forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00648] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 43, SEQ ID NO: 26, SEQ ID NO: 27, and SEQ ID NO: 46 are disclosed in Table 5. [00649] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. NAI-1540294631v3 189 [00650] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, and SEQ ID NO: 46 are disclosed in Table 6. [00651] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. [00652] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 113, SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, and SEQ ID NO: 118 are disclosed in Table 7. NAI-1540294631v3 190 [00653] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136. [00654] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 131, SEQ ID NO: 132, SEQ ID NO: 133, SEQ ID NO: 134, SEQ ID NO: 135, and SEQ ID NO: 136 are disclosed in Table 8. [00655] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: NAI-1540294631v3 191 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154. [00656] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 151, SEQ ID NO: 152, SEQ ID NO: 153, and SEQ ID NO: 154 are disclosed in Table 9. [00657] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. [00658] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence NAI-1540294631v3 192 set forth in SEQ ID NO: 169. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 131, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 134, SEQ ID NO: 135, and SEQ ID NO: 169 are disclosed in Table 10. [00659] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. [00660] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 41, SEQ ID NO: 24, SEQ ID NO: 182, SEQ ID NO: 26, SEQ ID NO: 27, and SEQ ID NO: 46 are disclosed in Table 11. [00661] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification thereof, and a CDR3 NAI-1540294631v3 193 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200. [00662] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 195, SEQ ID NO: 196, SEQ ID NO: 197, SEQ ID NO: 198, SEQ ID NO: 199, and SEQ ID NO: 200 are disclosed in Table 12. [00663] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. [00664] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and NAI-1540294631v3 194 a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 213, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 214, SEQ ID NO: 135, and SEQ ID NO: 169 are disclosed in Table 13. [00665] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232. [00666] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 227, SEQ ID NO: 228, SEQ ID NO: 229, SEQ ID NO: 230, SEQ ID NO: 231, and SEQ ID NO: 232 are disclosed in Table 14. [00667] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification NAI-1540294631v3 195 thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. [00668] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 113, SEQ ID NO: 245, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117, and SEQ ID NO: 118 are disclosed in Table 15. [00669] In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof. In certain embodiments, the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260. In certain embodiments, the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. NAI-1540294631v3 196 [00670] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and a VL comprising a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261. In certain embodiments, the CDRs are identified according to the Kabat numbering system. SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO: 260, SEQ ID NO: 230, SEQ ID NO: 231, and SEQ ID NO: 261 are disclosed in Table 16. [00671] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 29; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 30. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 29. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 30. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 29 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 30. [00672] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 47; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 48. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 47. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 48. In certain embodiments, the anti-GPC3 antibody or antigen- NAI-1540294631v3 197 binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 47 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 48. [00673] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 62; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 63. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 62. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 63. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 62 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 63. [00674] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 75; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 76. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 76. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 76. [00675] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 89; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at NAI-1540294631v3 198 least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 90. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 89. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 90. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 89 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 90. [00676] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 101; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 102. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 101. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 102. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 101 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 102. [00677] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 119; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 120. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 120. In certain embodiments, the anti-GPC3 antibody or antigen- NAI-1540294631v3 199 binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 120. [00678] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 137; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 138. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 138. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 138. [00679] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 155; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 156. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 156. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 156. [00680] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 170; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at NAI-1540294631v3 200 least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 171. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 170. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 171. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 170 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 171. [00681] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 183; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 184. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 184. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 184. [00682] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 201; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 202. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 202. In certain embodiments, the anti-GPC3 antibody or antigen- NAI-1540294631v3 201 binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 202. [00683] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 215; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 216. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 215. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 216. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 215 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 216. [00684] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 233; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 234. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 233. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 234. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 233 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 234. [00685] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 246; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at NAI-1540294631v3 202 least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 247. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 247. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 247. [00686] In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 262; and/or a VL comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% identical to the amino acid sequence set forth in SEQ ID NO: 263. In certain embodiments, the anti-GPC3 antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 262. In certain embodiments, the anti- GPC3 antibody or antigen-binding fragment thereof comprises a VL comprising the amino acid sequence set forth in SEQ ID NO: 263. In certain embodiments, the anti-GPC3 antibody or antigen- binding fragment thereof comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 262 and a VL comprising the amino acid sequence set forth in SEQ ID NO: 263. [00687] The antibody designated as “ATA001” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 1; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 1. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 29 (disclosed in Table 1) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 30 (disclosed in Table 1). [00688] The antibody designated as “ATA002” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 2; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 2. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 47 (disclosed in Table 2) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 48 (disclosed in Table 2). [00689] The antibody designated as “ATA003” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 3; and the VL CDR1, VL CDR2, and VL CDR3 sequences NAI-1540294631v3 203 disclosed in Table 3. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 62 (disclosed in Table 3) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 63 (disclosed in Table 3). [00690] The antibody designated as “ATA004” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 4; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 4. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 75 (disclosed in Table 4) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 76 (disclosed in Table 4). [00691] The antibody designated as “ATA005” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 5; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 5. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 89 (disclosed in Table 5) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 90 (disclosed in Table 5). [00692] The antibody designated as “ATA006” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 6; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 6. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 101 (disclosed in Table 6) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 102 (disclosed in Table 6). [00693] The antibody designated as “ATA007” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 7; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 7. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 119 (disclosed in Table 7) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 120 (disclosed in Table 7). [00694] The antibody designated as “ATA008” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 8; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 8. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 137 (disclosed in Table 8) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 138 (disclosed in Table 8). [00695] The antibody designated as “ATA009” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 9; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 9. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 155 (disclosed in Table 9) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 156 (disclosed in Table 9). NAI-1540294631v3 204 [00696] The antibody designated as “ATA010” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 10; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 10. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 170 (disclosed in Table 10) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 171 (disclosed in Table 10). [00697] The antibody designated as “ATA011” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 11; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 11. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 183 (disclosed in Table 11) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 184 (disclosed in Table 11). [00698] The antibody designated as “ATA012” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 12; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 12 The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 201 (disclosed in Table 12) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 202 (disclosed in Table 12). [00699] The antibody designated as “ATA013” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 13; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 13. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 215 (disclosed in Table 13) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 216 (disclosed in Table 13). [00700] The antibody designated as “ATA014” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 14; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 14. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 233 (disclosed in Table 14) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 234 (disclosed in Table 14). [00701] The antibody designated as “ATA015” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 15; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 15. The ATA001 antibody comprises a VH comprising the amino acid sequence set forth in SEQ ID NO: 246 (disclosed in Table 15) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 247 (disclosed in Table 15). [00702] The antibody designated as “ATA016” comprises the VH CDR1, VH CDR2, and VH CDR3 sequences disclosed in Table 16; and the VL CDR1, VL CDR2, and VL CDR3 sequences disclosed in Table 16. The ATA001 antibody comprises a VH comprising the amino acid sequence NAI-1540294631v3 205 set forth in SEQ ID NO: 262 (disclosed in Table 16) and a VL comprising the amino acid sequence set forth in SEQ ID NO: 263 (disclosed in Table 16). [00703] In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof comprises a fragment crystallizable region (Fc region). Fc region is the C-terminal region of an immunoglobulin heavy chain. The Fc region can be a functional Fc region, a native Fc region, a recombinant Fc region, or a variant Fc region. In certain embodiments, the Fc region comprises one or more heavy chain constant domains (e.g., CH2, and CH3). In certain embodiments, the variant Fc region comprises one or more modifications (e.g., substitutions) in the CH2 domain. In certain embodiments, the variant Fc region comprises one or more modifications (e.g., substitutions) in the CH3 domain. In certain embodiments, the heavy chain constant region is chosen from IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE. In certain embodiments, the heavy chain constant region is chosen from IgG1, IgG2, IgG3, and IgG4. In certain embodiments, the heavy chain constant region is chosen from IgG1. In certain embodiments, the heavy chain constant region is chosen from human IgG1. [00704] In certain embodiments, the Fc region is an IgG1 Fc region. In certain embodiments, the Fc region is a human IgG1 Fc region. In certain embodiments, the Fc region is a native human IgG1 Fc region. In certain embodiments, the Fc region comprises the amino acid sequence set forth in SEQ ID NO: 357. SEQ ID NO: 357 is provided below. DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPS RDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK [SEQ ID NO: 357] [00705] In certain embodiments, the Fc region comprises an amino acid sequence that is at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to the amino acid sequence set forth in SEQ ID NO: 357. [00706] A “functional Fc region” possesses an “effector function” of a native sequence Fc region. Exemplary “effector functions” include C1q binding; complement dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; down regulation of cell surface receptors (e.g., B cell receptor; BCR), etc. Such effector functions generally require the Fc region to be combined with a binding region or binding domain (e.g., an antibody variable region or domain) and can be assessed using various assays as disclosed. NAI-1540294631v3 206 [00707] A “native Fc region” encompasses a Fc region found in nature, and not manipulated, modified, and/or changed (e.g., isolated, purified, selected, including or combining with other sequences such as variable region sequences) by a human. Non-limiting examples of native human Fc regions include a native human IgG1 Fc region (non-A and A allotypes), a native human IgG2 Fc region, a native human IgG3 Fc region a native human IgG4 Fc region as well as naturally occurring variants of the foregoing. [00708] A “variant Fc region” comprises an amino acid sequence which differs from that of a native sequence Fc region by virtue of at least one amino acid modification, (e.g., substituting, addition, or deletion) preferably one or more amino acid substitution(s). In certain embodiments, the variant Fc region comprises at least one amino acid substitution compared to a native Fc region or to the Fc region of a parent polypeptide, e.g., from about 1 to about 10 amino acid substitutions, from about 1 to about 5 amino acid substitutions in a native sequence Fc region or in the Fc region of the parent polypeptide. The variant Fc region described herein can possess at least about 80%, at least about 90%, at least about 95% or at least about 99% sequence identity with a native Fc region and/or with an Fc region of a parent polypeptide. A variant Fc region may comprises a loss of effector function (e.g., silent Fc). [00709] The constant region/framework region of the presently disclosed antibodies can be altered, for example, by amino acid substitution, to modify the properties of the antibody (e.g., to increase or decrease one or more of: antigen binding affinity, Fc receptor binding, antibody carbohydrate, for example, glycosylation, fucosylation, etc. , the number of cysteine residues, effector cell function, effector cell function, complement function or introduction of a conjugation site). [00710] In certain embodiments, the presently disclosed anti-GPC3 antibody is a human antibody. In certain embodiments, the presently disclosed anti-GPC3 antibody is a humanized antibody. In certain embodiments, the presently disclosed anti-GPC3 antibody is a monoclonal antibody. In certain embodiments, the presently disclosed anti-GPC3 antibody is a human monoclonal antibody. In certain embodiments, the presently disclosed anti-GPC3 antibody is a humanized monoclonal antibody. [00711] In certain embodiments, the presently disclosed antigen-binding fragment is a Fab, Fab’, F(ab’)2, Fv, or single chain variable fragment (scFv). In certain embodiments, a presently disclosed antigen-binding fragment is an scFv. [00712] In certain embodiments, the presently disclosed antigen-binding fragment is an scFv-Fc. A scFv-Fc fragment comprises an scFv that is connected to a Fc region. NAI-1540294631v3 207 5.5.2. Multispecific Molecules [00713] The presently disclosed subject matter further provides for multispecific molecules comprising an anti-GPC3 antibody or an antigen-binding portion thereof disclosed herein. In certain embodiments, the multispecific molecules are multispecific antibodies. In certain embodiments, the multispecific molecules (e.g., multispecific antibodies) are bispecific molecules (e.g., bispecific antibodies). [00714] In certain embodiments, the multispecific molecule binds to at least two different binding sites or target molecules. In certain embodiments, the multispecific molecule comprises at least a first binding specificity for a GPC3 integrin, and a second binding specificity for a second target epitope. The second target epitope can be a GPC3 epitope, or a non-GPC3 epitope, e.g., an epitope on a second target antigen different from GPC3. In certain embodiments, the non-GPC3 epitope is the second target antigen is tumor antigen. [00715] The presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof can be derivatized or linked to more than one other functional molecule to generate multispecific molecules. To create a multispecific molecule, a presently disclosed anti-GPC3 antibody or an antigen-binding fragment thereof can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other binding molecules, such as another antibody, antibody fragment, peptide or binding mimetic, such that a multispecific molecule results. [00716] The multispecific molecules of the presently disclosed subject matter can be prepared by conjugating the constituent binding specificities using methods known in the art. For example, each binding specificity of the bispecific molecule can be generated separately and then conjugated to one another. When the binding specificities are proteins or peptides, a variety of coupling or cross- linking agents can be used for covalent conjugation. Non-limiting examples of cross-linking agents include protein A, carbodiimide, N-succinimidyl-S-acetyl-thioacetate (SAT A), 5, 5'-dithiobis(2- nitrobenzoic acid) (DTNB), o-phenylenedimaleimide (oPDM), N-succinimidyl-3-(2- pyridyldithio)propionate (SPDP), and sulfosuccinimidyl 4-(N-maleimidom ethyl) cyclohaxane-l- carboxylate (sulfo-SMCC). [00717] Other formats of bispecific antibodies can be constructed, such tandem scFv molecules (taFv), diabodies (Db), or single chain diabodies (scDb), and fusion protein with human serum albumin (Ryutaro, et al., J Biol Chem 2011; 286: 1812-1818; Anja, et al., Blood 2000; 95(6): 2098- 2103; Weiner, et al., J. Immunology 1994; 152(5): 2385-2392; Dafne, et al., J Biol Chem 2007; 282: 12650-12660), but are devoid of Fc effector functions with distinct pharmacokinetic profiles. NAI-1540294631v3 208 [00718] Methods for making multispecific molecules are known in the art, such as, by co- expression of two immunoglobulin heavy chain-light chain pairs, where the two heavy chains have different specificities. Exemplary structures of multispecific molecules are known in the art. [00719] For example, bispecific molecules can be classified into different structural groups: (i) bispecific immunoglobulin G (BsIgG); (ii) IgG appended with an additional antigen-binding moiety; (iii) bispecific antibody fragments; (iv) bispecific fusion proteins; and (v) bispecific antibody conjugates. In certain embodiments, BslgG comprises heavy chains that are engineered for heterodimerization. For example, heavy chains can be engineered for heterodimerization using a “knobs-into-holes” strategy, a SEED platform, a common heavy chain (e.g., in κλ-bodies), and use of heterodimeric Fc regions. Strategies are known in the art to avoid heavy chain pairing of homodimers in BsIgG, including knobs-into-holes, duobody, azymetric, charge pair, HA-TF, SEEDbody, and differential protein A affinity. [00720] Another bispecific molecule format is IgG appended with an additional antigen-binding moiety. For example, monospecific IgG can be engineered to have bi-specificity by appending an additional antigen-binding unit onto the monospecific IgG, for example, at the N- or C- terminus of either the heavy or light chain. [00721] Bispecific antibody fragments (BsAb) are a format of bispecific molecules that lack some or all of the antibody constant domains. For example, some BsAb lack an Fc region. In certain embodiments, bispecific antibody fragments include heavy and light chain regions that are connected by a peptide linker that permits efficient expression of the BsAb in a single host cell. Non-limiting examples of bispecific antibody fragments include, but are not limited to, nanobody, nanobody-HAS, BiTE, Diabody, DART, TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, triple body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab’)2, F(ab’)2-scFv2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, Diabody-Fc, tandem scFv-Fc, and intrabody. [00722] Bispecific fusion proteins include antibody fragments linked to other proteins. For example, bispecific fusion proteins can be linked to other proteins to add additional specificity and/or functionality. In certain embodiments, the dock-and-lock (DNL) method can be used to generate bispecific antibody molecules with higher valency. For example, bispecific antibody fusions to albumin binding proteins or human serum albumin can be extend the serum half-life of antibody fragments. In certain embodiments, chemical conjugation, for example, chemical conjugation of antibodies and/or antibody fragments, can be used to create BsAb molecules. NAI-1540294631v3 209 [00723] Methods of production of multispecific molecules, including bispecific molecules, are known in the art. For example, multispecific molecules, including bispecific molecules, can be produced by separate expression of the component antibodies in different host cells and subsequent purification/assembly or by expression of the component antibodies in a single host cell. Purification of multispecific (e.g., bispecific) molecules can be performed by various methods known in the art, including affinity chromatography. 5.5.3. Cross-competing Antibodies [00724] The presently disclosed subject matter further provides antibodies or antigen-binding fragments thereof that cross-compete for binding to a GPC3 antigen (e.g., a human GPC3 antigen) with any of the presently disclosed anti-GPC3 antibodies or antigen-binding fragments thereof (e.g., those disclosed in Sections 5.5.1 and 5.5.2, e.g., ATA001, ATA002, ATA003, ATA004, ATA005, ATA006, ATA007, ATA008, ATA009, ATA010, ATA011, ATA012, ATA013, ATA014, ATA015, and ATA016). For example, the cross-competing antibodies bind to the same epitope region, e.g., same epitope, adjacent epitope, or overlapping as any of the presently disclosed anti-GPC4 antibodies or antigen-binding fragments thereof (e.g., those disclosed in Section 5.5.1 and 5.5.2, e.g., ATA001, ATA002, ATA003, ATA004, ATA005, ATA006, ATA007, ATA008, ATA009, ATA010, ATA011, ATA012, ATA013, ATA014, ATA015, and ATA016). [00725] The competition can be determined by an assay in which the test antibody under study prevents or inhibits the specific binding of the reference antibody to a common epitope or a common antigen (e.g., a GPC3 antigen). Numerous types of competitive binding assays can be used to determine if a test antibody competes with a reference antibody for binding to α5 integrin (e.g., human GPC3). Examples of assays that can be employed include solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see, e.g., Stahli et al., (1983) Methods in Enzymology 9:242-253); solid phase direct biotin-avidin EIA (see, e.g., Kirkland et al., (1986) J. Immunol.137:3614-3619) solid phase direct labeled assay, solid phase direct labeled sandwich assay (see, e.g., Harlow and Lane, (1988) Antibodies, A Laboratory Manual, Cold Spring Harbor Press); solid phase direct label RIA using 1- 125 label (see, e.g., Morel et al., (1988) Molec. Immunol.25:7-15); solid phase direct biotin-avidin EIA (see, e.g., Cheung, et al., (1990) Virology 176:546-552); and direct labeled RIA (Moldenhauer et al., (1990) Scand. J. Immunol.32:77-82). Typically, such an assay involves the use of a purified antigen (e.g., α5 integrin, such as human α5 integrin) bound to a solid surface or cells bearing either of an un-labelled test antigen binding protein (e.g., test α5 integrin antibody) or a labeled reference antigen binding protein (e.g., reference anti-α5 integrin antibody). Competitive inhibition may be NAI-1540294631v3 210 measured by determining the amount of label bound to the solid surface or cells in the presence of the test antigen binding protein. Usually the test antigen binding protein is present in excess. Antibodies identified by competition assay (competing antibodies) include antibodies binding to the same epitope as the reference antibody and/or antibodies binding to an adjacent epitope sufficiently proximal to the epitope bound by the reference for antibodies steric hindrance to occur (e.g., similar epitope or overlapping epitope). Usually, when a competing antibody is present in excess, it will inhibit specific binding of a reference antibody to a common epitope or common antigen by at least about 20%, for example, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96% or at least about 97%, at least about 98%, or at least about 99% or more. 5.5.4. Immunoconjugates [00726] The presently disclosed subject further provides conjugates comprising a presently disclosed anti-GPC4 antibody or antigen-binding fragment thereof. In certain embodiments, the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof is conjugated to a therapeutic moiety, such as a cytotoxin, a drug (e.g., an immunosuppressant) or a radiotoxin. Such conjugates are referred to herein as “immunoconjugates.” Immunoconjugates that include one or more cytotoxins are referred to as “immunotoxins.” A cytotoxin or cytotoxic agent includes any agent that is detrimental to (e.g., kills) cells. Non-limiting examples include taxol (such as ricin, diphtheria, gelonin), cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof. Therapeutic agents also include, for example, calecheamicin, aureastatin, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepa chlorambucil, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis- dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti -mitotic agents (e.g., vincristine and vinblastine). [00727] Other non-limiting examples of therapeutic cytotoxins that can be conjugated to the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof include duocarmycins, NAI-1540294631v3 211 calicheamicins, maytansines and auristatins, and derivatives thereof. A non-limiting example of a calicheamicin antibody conjugate is commercially available (Mylotarg™; Wyeth-Ayerst). [00728] Cytoxins can be conjugated to the presently disclosed anti-GPC3 antibody or antigen- binding fragment thereof using linker technology available in the art. Examples of linker types that have been used to conjugate a cytotoxin to an antibody include, but are not limited to, hydrazones, thioethers, esters, disulfides and peptide-containing linkers. A linker can be chosen that is, for example, susceptible to cleavage by low pH within the lysosomal compartment or susceptible to cleavage by proteases, such as proteases preferentially expressed in tumor tissue such as cathepsins (e.g., cathepsins B, C, D). [00729] The presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof can also be conjugated to a radioactive isotope to generate cytotoxic radiopharmaceuticals, also referred to as radioimmunoconjugates. Examples of radioactive isotopes that can be conjugated to antibodies for use diagnostically or therapeutically include, but are not limited to, Y, I, Ac, Bi, Ra, Lu, and Th. Method for preparing radioimmunoconjugates are established in the art. Examples of radioimmunoconjugates are commercially available, including Zevalin™ (IDEC Pharmaceuticals) and Bexxar™ (Corixa Pharmaceuticals), and similar methods can be used to prepare radioimmunoconjugates using the presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof. [00730] The conjugates can be used to modify a given biological response, and the drug moiety is not to be construed as limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, an enzymatically active toxin, or active fragment thereof, such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor (TNF) or interferon-γ; or, biological response modifiers such as, for example, lymphokines, interleukin-1 (“IL-1”), interleukin-2 (“IL-2”), interleukin-6 (“IL-6”), granulocyte macrophage colony stimulating factor (“GM-CSF”), granulocyte colony stimulating factor (“G-CSF”), or other growth factors. 5.5.5. Engineered and Modified Antibodies [00731] The presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof can be prepared using an antibody or an antigen-binding fragment thereof comprising one or more of the V and/or V sequences disclosed herein as starting material to engineer a modified antibody, which modified antibody may have altered properties from the starting antibody. An antibody can be engineered by modifying one or more residues within one or both variable regions (i.e., V and/or V), for example within one or more CDR regions and/or within one or more framework regions. NAI-1540294631v3 212 Additionally or alternatively, an antibody can be engineered by modifying residues within the constant region(s), for example to alter the effector function(s) of the antibody. [00732] One type of variable region engineering that can be performed is CDR grafting. [00733] Antibodies interact with target antigens predominantly through amino acid residues that are located in the six heavy and light chain CDRs. For this reason, the amino acid sequences within CDRs are more diverse between individual antibodies than sequences outside of CDRs. Because CDR sequences are responsible for most antibody-antigen interactions, it is possible to express recombinant antibodies that mimic the properties of specific naturally occurring antibodies by constructing expression vectors that include CDR sequences from the specific naturally occurring antibody grafted onto framework sequences from a different antibody with different properties (see, e.g., Riechmann, L. et al. (1998) Nature 332:323-327; Jones, P. et al. (1986) Nature 321 :522-525; Queen, C. et al. (1989) Proc. Natl. Acad. See. U.S.A.86: 10029-10033; U.S. Patent No.5,225,539 to Winter, and U.S. Patent Nos.5,530,101; 5,585,089; 5,693,762 and 6,180,370 to Queen et al.) [00734] Framework sequences can be obtained from public DNA databases or published references that include germline antibody gene sequences. The V CDRl, CDR2, and CDR3 sequences, and the V CDR1, CDR2, and CDR3 sequences, can be grafted onto framework regions that have the identical sequence as that found in the germline immunoglobulin gene from which the framework sequence derive, or the CDR sequences can be grafted onto framework regions that contain one or more mutations as compared to the germline sequences. [00735] Another type of variable region modification is to mutate amino acid residues within the V and/or V CDRl, CDR2 and/or CDR3 regions to thereby improve one or more binding properties (e.g., affinity) of the antibody of interest. Site-directed mutagenesis or PCR-mediated mutagenesis can be performed to introduce the mutation(s) and the effect on antibody binding, or other functional property of interest, can be evaluated in in vitro or in vivo assays. In certain embodiments, conservative modifications are introduced. The mutations may be amino acid substitutions, additions or deletions. For example, no more than one, two, three, four or five residues within a CDR region are altered. [00736] Accordingly, the presently disclosed subject matter provides for anti-GPC3 antibodies or antigen-binding fragments thereof comprising a V comprising: (a) a CDR1 sequence of the antibodies and antigen-binding fragments thereof disclosed herein, or an amino acid sequence having at least one (e.g., no more than one, no more than two, no more than three, no more than four or no more than five) amino acid modification (e.g., substitution, deletion and/or addition) as compared to the V CDR1 sequence of any one of the antibodies or antigen-binding fragments NAI-1540294631v3 213 thereof disclosed herein; (b) a CDR2 sequence of any one of the antibodies or antigen-binding fragments thereof disclosed herein, or an amino acid sequence having at least one (e.g., no more than one, no more than two, no more than three, no more than four or no more than five) amino acid modification (e.g., substitution, deletion and/or addition) as compared to the V CDR2 of any one of the antibodies or antigen-binding fragments thereof disclosed herein; and (c) a CDR3 sequence of any one of the antibodies or antigen-binding fragments thereof disclosed herein, or an amino acid sequence having at least one (e.g., no more than one, no more than two, no more than three, no more than four or no more than five) amino acid modification (e.g., substitution, deletion and/or addition) as compared to the V CDR3 of any one of the antibodies or antigen-binding fragments thereof disclosed herein; and a V comprising (a) a CDR1 sequence of any one of the antibodies or antigen- binding fragments thereof disclosed herein, or an amino acid sequence having at least one (e.g., no more than one, no more than two, no more than three, no more than four or no more than five) amino acid modification (e.g., substitution, deletion and/or addition) as compared to the VL CDR1 of any one of the antibodies or antigen-binding fragments thereof disclosed herein; (b) a CDR2 sequence of any one of the antibodies or antigen-binding fragments thereof disclosed herein, or an amino acid sequence having at least one (e.g., no more than one, no more than two, no more than three, no more than four or no more than five) amino acid modification (e.g., substitution, deletion and/or addition) as compared to the VL CDR2 of any one of the antibodies or antigen-binding fragments thereof disclosed herein; and/or (c) a CDR3 sequence of any one of the antibodies or antigen-binding fragments thereof disclosed herein, or an amino acid sequence having at least one (e.g., no more than one, no more than two, no more than three, no more than four or no more than five) amino acid modification (e.g., substitution, deletion and/or addition) as compared to the VL CDR3 of any one of the antibodies or antigen-binding fragments thereof disclosed herein. [00737] Engineered antibodies of the presently disclosed subject matter include those in which modifications are made to framework residues within VH and/or VL, e.g., to improve the properties of the antibody. Typically such framework modifications are made to decrease the immunogenicity of the antibody. For example, one approach is to “backmutate” one or more framework residues to the corresponding germline sequence. More specifically, an antibody that has undergone somatic mutation may contain framework residues that differ from the germline sequence from which the antibody is derived. Such residues can be identified by comparing the antibody framework sequences to the germline sequences from which the antibody is derived. [00738] In addition or alternative to modifications made within the framework or CDR regions, the presently disclosed anti-GPC3 antibodies or antigen-binding fragments thereof can be engineered NAI-1540294631v3 214 to include modifications within the Fc region, typically to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and/or antigen- dependent cellular cytotoxicity. Furthermore, a presently disclosed anti-GPC3 antibody or antigen- binding fragment thereof can be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or be modified to alter its glycosylation, again to alter one or more functional properties of the antibody. The hinge region of CH1 can be modified such that the number of cysteine residues in the hinge region is altered, e.g., increased or decreased. The number of cysteine residues in the hinge region of CH1 is altered to, for example, facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody. The Fc hinge region of an antibody may be mutated to decrease the biological half life of the antibody. More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of the Fc-hinge fragment such that the antibody has impaired Staphylococcyl protein A (SpA) binding relative to native Fc-hinge domain SpA binding. The antibody can be modified to increase its biological half-life, e.g., the antibody can be altered within the CH1 or CL region to contain a salvage receptor binding epitope taken from two loops of a CH2 domain of an Fc region of an IgG. Furthermore, the Fc region can be altered by replacing at least one amino acid residue with a different amino acid residue to alter the effector function(s) of the antibody. The Fc region can be modified to increase the ability of the antibody to mediate antibody dependent cellular cytotoxicity (ADCC) and/or to increase the affinity of the antibody for an Fcγ receptor. In certain embodiments, a presently disclosed anti-GPC3 antibody comprises an afucosylated Fc region. Removal of the fucose residue from the N-glycans of the Fc portion of immunoglobulin G (IgG) can result in a dramatic enhancement of ADCC through improved affinity for Fcγ receptor Ilia (FcγRIIIa). [00739] Additionally or alternatively, the glycosylation of a presently disclosed anti-GPC3 antibody can be modified. For example, an aglycoslated antibody can be made (i.e., the antibody lacks glycosylation). Glycosylation can be altered to, for example, increase the affinity of the antibody for antigen. Such carbohydrate modifications can be accomplished by, for example, altering one or more sites of glycosylation within the antibody sequence. For example, one or more amino acid substitution can be made that result in elimination of one or more variable region framework glycosylation sites to thereby eliminate glycosylation at that site. [00740] Additionally or alternatively, an antibody can be made that has an altered type of glycosylation, such as a hypofucosylated antibody having reduced amounts of fucosyl residues or an antibody having increased bisecting GlcNac structures. Such altered glycosylation patterns have been demonstrated to increase the ADCC ability of antibodies. Such carbohydrate modifications can NAI-1540294631v3 215 be accomplished by, for example, expressing the antibody in a host cell with altered glycosylation machinery. [00741] Another modification of the antibodies may be pegylation. An antibody can be pegylated to, for example, increase the biological (e.g., serum) half-life of the antibody. In certain embodiments, to pegylate an antibody, the antibody, or fragment thereof, typically is reacted with polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions in which one or more PEG groups become attached to the antibody or antibody fragment. The pegylation may be carried out via an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer). As used herein, the term "polyethylene glycol" is intended to encompass any of the forms of PEG that have been used to derivatize other proteins, such as mono (Ci-Ci) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol- maleimide. The antibody to be pegylated may be an aglycosylated antibody. 5.6. Nucleic Acids and Vectors [00742] The presently disclosed subject matter provides nucleic acids encoding the presently disclosed binding molecules. In one aspect, the presently disclosed subject matter provides GPC3- targeted CARs (e.g., those disclosed in Section 5.3), and cells comprising such nucleic acids. In certain embodiments, the nucleic acid further comprises a promoter that is operably linked to the nucleic acid encoding a presently disclosed GPC3-targeted CAR. [00743] In another aspect, the presently disclosed subject matter provides nucleic acids encoding the anti-GPC3 antibodies or antigen-binding fragments thereof disclosed herein (e.g., those disclosed in Section 5.4) or a region thereof, and cells comprising such nucleic acids. In certain embodiments, the nucleic acid comprises a first polynucleotide encoding an anti-GPC3 VH disclosed herein (e.g., one disclosed in Sections 5.3 and 5.4, e.g., one disclosed in Tables 1-16). In certain embodiments, the first polynucleotide encodes an anti-GPC3 VH comprising the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262. In certain embodiments, the first polynucleotide encodes an anti-GPC3 VH comprising a CDR1, a CDR2, and a CDR3 disclosed in Tables 1-16. [00744] In certain embodiments, the nucleic acid comprises a second polynucleotide encoding an anti-GPC3 VL disclosed herein (e.g., one disclosed in Sections 5.3 and 5.4, e.g., one disclosed in Tables 1-16). In certain embodiments, the second polynucleotide encodes an anti-GPC3 VL comprising the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, NAI-1540294631v3 216 SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263. In certain embodiments, the second polynucleotide encodes an anti-GPC3 VL comprising a CDR1, a CDR2, and a CDR3 disclosed in Tables 1-16. [00745] In certain embodiments, the nucleic acid further comprises a promoter that is operably linked to the nucleic acid encoding a presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof, or a region of a presently disclosed anti-GPC3 antibody or antigen-binding fragment thereof (e.g., VH or VL). [00746] In certain embodiments, the promoter is endogenous or exogenous. In certain embodiments, the exogenous promoter is selected from the group consisting of an elongation factor (EF)-1 promoter, a cytomegalovirus immediate-early promoter (CMV) promoter, a simian virus 40 early promoter (SV40) promoter, a phosphoglycerate kinase (PGK) promoter, a metallothionein promoter, and Ubiquitin C promoter. In certain embodiments, the endogenous promoter is selected from a TCR alpha promoter, a TCR beta promoter, and a beta 2-microglobulin promoter. In certain embodiments, the promoter is an inducible promoter. In certain embodiment, the inducible promoter is selected from the group consisting of a NFAT transcriptional response element (TRE) promoter, a CD69 promoter, a CD25 promoter, an IL-2 promoter, a 4-1BB promoter, a PD1 promoter, and a LAG3 promoter. [00747] The presently disclosed subject matter also provides vectors comprising the presently disclosed nucleic acids. In certain embodiments, the vector is an expression vector. [00748] The nucleic acids can be delivered into cells by art-known methods or as described herein. Genetic modification of a cell can be accomplished by transducing a substantially homogeneous cell composition with a recombinant DNA construct. In certain embodiments, a retroviral vector (e.g., gammaretroviral vector or lentiviral vector) is employed for the introduction of the DNA construct into the cell. For example, a presently disclosed nucleic acid can be cloned into a retroviral vector and expression can be driven from its endogenous promoter, from the retroviral long terminal repeat, or from a promoter specific for a target cell type of interest. [00749] Expression of nucleic acids encoding CARs is typically achieved by operably linking a nucleic acid encoding the CAR polypeptide to a promoter and incorporating the construct into an expression vector. Typical cloning vectors contain transcription and translation terminators, initiation sequences, and promoters useful for regulation of the expression of the desired nucleic acid sequence. NAI-1540294631v3 217 [00750] The disclosed nucleic acid can be cloned into a number of types of vectors. For example, the nucleic acid can be cloned into a vector including, but not limited to a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid. Vectors of particular interest include expression vectors, replication vectors, probe generation vectors, and sequencing vectors. [00751] Further, the expression vector may be provided to a cell in the form of a viral vector. Viral vector technology is well known in the art and is described, for example, in Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and in other virology and molecular biology manuals. Viruses, which are useful as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, and lentiviruses. In general, a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers. In some embodiments, the polynucleotide vectors are lentiviral or retroviral vectors. [00752] A number of viral based systems have been developed for gene transfer into mammalian cells. For example, retroviruses provide a convenient platform for gene delivery systems. A selected gene can be inserted into a vector and packaged in retroviral particles using techniques known in the art. The recombinant virus can then be isolated and delivered to cells of the subject either in vivo or ex vivo. [00753] One example of a suitable promoter is the immediate early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strong constitutive promoter sequence capable of driving high levels of expression of any polynucleotide sequence operatively linked thereto. Another example of a suitable promoter is Elongation Growth Factor-1α (EF-1α). However, other constitutive promoter sequences may also be used, including, but not limited to the simian virus 40 (SV40) early promoter, MND (myeloproliferative sarcoma virus) promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) long terminal repeat (LTR) promoter, MoMuLV promoter, an avian leukemia virus promoter, an Epstein-Barr virus immediate early promoter, a Rous sarcoma virus promoter, as well as human gene promoters such as, but not limited to, the actin promoter, the myosin promoter, the hemoglobin promoter, and the creatine kinase promoter. The promoter can alternatively be an inducible promoter. Examples of inducible promoters include, but are not limited to a metallothionine promoter, a glucocorticoid promoter, a progesterone promoter, and a tetracycline promoter. [00754] Additional promoter elements, e.g., enhancers, regulate the frequency of transcriptional initiation. Typically, these are located in the region 30-110 bp upstream of the start site, although a NAI-1540294631v3 218 number of promoters have recently been shown to contain functional elements downstream of the start site as well. The spacing between promoter elements frequently is flexible, so that promoter function is preserved when elements are inverted or moved relative to one another. [00755] In order to assess the expression of a CAR polypeptide or portions thereof, the expression vector to be introduced into a cell can also contain either a selectable marker gene or a reporter gene or both to facilitate identification and selection of expressing cells from the population of cells sought to be transfected or infected through viral vectors. In other aspects, the selectable marker may be carried on a separate piece of DNA and used in a co-transfection procedure. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers include, for example, antibiotic-resistance genes. [00756] Reporter genes are used for identifying potentially transfected cells and for evaluating the functionality of regulatory sequences. In general, a reporter gene is a gene that is not present in or expressed by the recipient organism or tissue and that encodes a polypeptide whose expression is manifested by some easily detectable property, e.g., enzymatic activity. Expression of the reporter gene is assayed at a suitable time after the DNA has been introduced into the recipient cells. Suitable reporter genes may include genes encoding luciferase, beta-galactosidase, chloramphenicol acetyl transferase, secreted alkaline phosphatase, or the green fluorescent protein gene. Suitable expression systems are well known and may be prepared using known techniques or obtained commercially. In general, the construct with the minimal 5′ flanking region showing the highest level of expression of reporter gene is identified as the promoter. Such promoter regions may be linked to a reporter gene and used to evaluate agents for the ability to modulate promoter-driven transcription. [00757] Methods of introducing and expressing genes into a cell are known in the art. In the context of an expression vector, the vector can be readily introduced into a host cell, e.g., mammalian, bacterial, yeast, or insect cell by any method in the art. For example, the expression vector can be transferred into a host cell by physical, chemical, or biological means. [00758] Physical methods for introducing a polynucleotide into a host cell include calcium phosphate precipitation, lipofection, particle bombardment, microinjection, electroporation, and the like. Methods for producing cells comprising vectors and/or exogenous nucleic acids are well-known in the art. See, for example, Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York). NAI-1540294631v3 219 [00759] Biological methods for introducing a polynucleotide of interest into a host cell include the use of DNA and RNA vectors. Viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mammalian, e.g., human cells. [00760] Chemical means for introducing a polynucleotide into a host cell include colloidal dispersion systems, such as macromolecule complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system for use as a delivery vehicle in vitro and in vivo is a liposome (e.g., an artificial membrane vesicle). [00761] In the case where a non-viral delivery system is utilized, an exemplary delivery vehicle is a liposome. In another aspect, the nucleic acid may be associated with a lipid. The nucleic acid associated with a lipid may be encapsulated in the aqueous interior of a liposome, interspersed within the lipid bilayer of a liposome, attached to a liposome via a linking molecule that is associated with both the liposome and the oligonucleotide, entrapped in a liposome, complexed with a liposome, dispersed in a solution containing a lipid, mixed with a lipid, combined with a lipid, contained as a suspension in a lipid, contained or complexed with a micelle, or otherwise associated with a lipid. Lipid, lipid/DNA or lipid/expression vector associated compositions are not limited to any particular structure in solution. For example, they may be present in a bilayer structure, as micelles, or with a “collapsed” structure. They may also simply be interspersed in a solution, possibly forming aggregates that are not uniform in size or shape. Lipids are fatty substances which may be naturally occurring or synthetic lipids. For example, lipids include the fatty droplets that naturally occur in the cytoplasm as well as the class of compounds which contain long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, amino alcohols, and aldehydes. Lipids suitable for use can be obtained from commercial sources. For example, dimyristyl phosphatidylcholine (“DMPC”) can be obtained from Sigma, St. Louis, Mo.; dicetyl phosphate (“DCP”) can be obtained from K & K Laboratories (Plainview, N.Y.); cholesterol (“Choi”) can be obtained from Calbiochem-Behring; dimyristyl phosphatidylglycerol (“DMPG”) and other lipids may be obtained from Avanti Polar Lipids, Inc, (Birmingham, Ala.). [00762] Any targeted genome editing methods can also be used to deliver a presently disclosed GPC3-targeted CAR to a cell. In certain embodiments, a CRISPR system is used to deliver a presently disclosed GPC3-targeted CAR. In certain embodiments, zinc-finger nucleases are used to deliver a presently disclosed GPC3-targeted CAR. In certain embodiments, a TALEN system is used to deliver a presently disclosed GPC3-targeted CAR. NAI-1540294631v3 220 [00763] Clustered regularly-interspaced short palindromic repeats (CRISPR) system is a genome editing tool discovered in prokaryotic cells. When utilized for genome editing, the system includes Cas9 (a protein able to modify DNA utilizing crRNA as its guide), CRISPR RNA (crRNA, contains the RNA used by Cas9 to guide it to the correct section of host DNA along with a region that binds to tracrRNA (generally in a hairpin loop form) forming an active complex with Cas9), trans- activating crRNA (tracrRNA, binds to crRNA and forms an active complex with Cas9), and an optional section of DNA repair template (DNA that guides the cellular repair process allowing insertion of a specific DNA sequence). CRISPR/Cas9 often employs a plasmid to transfect the target cells. The crRNA needs to be designed for each application as this is the sequence that Cas9 uses to identify and directly bind to the target DNA in a cell. The repair template carrying CAR expression cassette need also be designed for each application, as it must overlap with the sequences on either side of the cut and code for the insertion sequence. Multiple crRNA’s and the tracrRNA can be packaged together to form a single-guide RNA (sgRNA). This sgRNA can be joined together with the Cas9 gene and made into a plasmid in order to be transfected into cells. [00764] A zinc-finger nuclease (ZFN) is an artificial restriction enzyme, which is generated by combining a zinc finger DNA-binding domain with a DNA-cleavage domain. A zinc finger domain can be engineered to target specific DNA sequences which allows a zinc-finger nuclease to target desired sequences within genomes. The DNA-binding domains of individual ZFNs typically contain a plurality of individual zinc finger repeats and can each recognize a plurality of base pairs. The most common method to generate new zinc-finger domain is to combine smaller zinc-finger “modules” of known specificity. The most common cleavage domain in ZFNs is the non-specific cleavage domain from the type IIs restriction endonuclease FokI. Using the endogenous homologous recombination (HR) machinery and a homologous DNA template carrying CAR expression cassette, ZFNs can be used to insert the CAR expression cassette into genome. When the targeted sequence is cleaved by ZFNs, the HR machinery searches for homology between the damaged chromosome and the homologous DNA template, and then copies the sequence of the template between the two broken ends of the chromosome, whereby the homologous DNA template is integrated into the genome. [00765] Transcription activator-like effector nucleases (TALEN) are restriction enzymes that can be engineered to cut specific sequences of DNA TALEN system operates on almost the same principle as ZFNs. They are generated by combining a transcription activator-like effectors DNA- binding domain with a DNA cleavage domain. Transcription activator-like effectors (TALEs) are composed of 33-34 amino acid repeating motifs with two variable positions that have a strong NAI-1540294631v3 221 recognition for specific nucleotides. By assembling arrays of these TALEs, the TALE DNA-binding domain can be engineered to bind desired DNA sequence, and thereby guide the nuclease to cut at specific locations in genome.cDNA expression for use in polynucleotide therapy methods can be directed from any suitable promoter (e.g., the human cytomegalovirus (CMV), simian virus 40 (SV40), metallothionein promoters, or Ubiquitin C promoter), and regulated by any appropriate mammalian regulatory element or intron (e.g. the elongation factor 1a enhancer/promoter/intron structure). For example, if desired, enhancers known to preferentially direct gene expression in specific cell types can be used to direct the expression of a nucleic acid. The enhancers used can include, without limitation, those that are characterized as tissue- or cell-specific enhancers. Alternatively, if a genomic clone is used as a therapeutic construct, regulation can be mediated by the cognate regulatory sequences or, if desired, by regulatory sequences derived from a heterologous source, including any of the promoters or regulatory elements described above. [00766] Methods for delivering the genome editing agents/systems can vary depending on the need. In certain embodiments, the components of a selected genome editing method are delivered as DNA constructs in one or more plasmids. In certain embodiments, the components are delivered via viral vectors. Common delivery methods include but is not limited to, electroporation, microinjection, gene gun, impalefection, hydrostatic pressure, continuous infusion, sonication, magnetofection, adeno-associated viruses, envelope protein pseudotyping of viral vectors, replication-competent vectors cis and trans-acting elements, herpes simplex virus, and chemical vehicles (e.g., oligonucleotides, lipoplexes, polymersomes, polyplexes, dendrimers, inorganic Nanoparticles, and cell-penetrating peptides). 5.7. Methods of Detection [00767] The present disclosure provides methods for detecting GPC3 in a whole cell or tissue. In certain embodiments, the method comprises: a) contacting a cell or tissue with an anti-GPC3 antibody or antigen-binding fragment thereof disclosed herein, wherein the anti-GPC3 antibody or antigen-binding fragment thereof comprises a detectable label; and b) determining the amount of the labeled anti-GPC3 antibody or antigen-binding fragment thereof bound to the cell or tissue. In certain embodiments, the method is for detecting human GPC3. [00768] In certain embodiments, b) determining the amount of the labeled anti-GPC3 antibody or antigen-binding fragment thereof bound to the cell or tissue comprises measuring the amount of detectable label associated with the cell or tissue, wherein the amount of bound antibody or antigen- binding fragment thereof indicates the amount of GPC3 (e.g., human GPC3) in the cell or tissue. NAI-1540294631v3 222 [00769] The cell or tissue can be any cell or tissue, including any normal, healthy, abnormal, tumor, or cancer cells and tissues. [00770] The present disclosure also provides methods for detecting or diagnosing an GPC3- associated disease, disorder, or condition. A more definitive diagnosis of a GPC3-associated disease, disorder, or condition may allow health professionals to employ preventative measures or aggressive treatment earlier thereby preventing the development or further progression of the α5 integrin-associated disease, disorder, or condition. In certain embodiments, the method comprises: (a) measuring an expression level of GPC3 in a cell or a tissue sample of a subject using an anti- GPC3 antibody or antigen-binding fragment thereof disclosed herein; and (b) comparing the expression level of GPC3 measured in (a) with a control expression level of GPC3, wherein an increase in the expression level of GPC3 measured in (a) as compared to the control expression level of GPC3 is indicative of an GPC3-associated disease, disorder, or condition. In certain embodiments, the control expression level of GPC3 is an expression level of GPC3 in a cell or a tissue sample of a subject not suffering from an GPC3-associated disease, disorder, or condition. 5.8. Formulations and Administration [00771] In one aspect, the presently disclosed subject matter provides compositions comprising the presently disclosed cells comprising a presently disclosed GPC3-targeted CAR. In another aspect, the presently disclosed subject matter provides compositions comprising the anti-GPC3 antibodies or antigen-binding fragments thereof, the conjugates, or the multispecific molecules disclosed herein. [00772] In certain embodiments, the composition is a pharmaceutical composition that further comprises a pharmaceutically acceptable carrier. [00773] The presently disclosed compositions can be conveniently provided as sterile liquid preparations, e.g., isotonic aqueous solutions, suspensions, emulsions, dispersions, or viscous compositions, which may be buffered to a selected pH. Liquid preparations are normally easier to prepare than gels, other viscous compositions, and solid compositions. Additionally, liquid compositions are somewhat more convenient to administer, especially by injection. Viscous compositions, on the other hand, can be formulated within the appropriate viscosity range to provide longer contact periods with specific tissues. Liquid or viscous compositions can comprise carriers, which can be a solvent or dispersing medium containing, for example, water, saline, phosphate buffered saline, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like) and suitable mixtures thereof. NAI-1540294631v3 223 [00774] Compositions comprising the presently disclosed cells can be provided systemically or directly to a subject in need thereof. In certain embodiments, the presently disclosed compositions are directly injected into an organ of interest. Alternatively, the presently disclosed compositions are provided indirectly to the organ of interest, for example, by administration into the circulatory system (e.g., the tumor vasculature). Expansion and differentiation agents can be provided prior to, during or after administration of compositions to increase production of cells in vitro or in vivo. [00775] The quantity of cells comprising the GPC3-targeted CAR comprised in the presently disclosed compositions can vary for the subject being treated. In certain embodiments, the presently disclosed composition comprises between about 1×104 and about 1×1010, between about 1×104 and about 1×107, between about 1×105 and about 1×107, between about 1×105 and about 1×108, between about 1×105 and about 109, between about 1×106 and about 1×108, between about 1×106 and about 1×109 , or between about 1×106 and about 1×1010 of the presently disclosed cells comprising the GPC3-targeted CAR. In certain embodiments, the presently disclosed composition comprises between about 1×105 and about 5×108 of the presently disclosed cells comprising the GPC3-targeted CAR. In certain embodiments, the presently disclosed composition comprises between about 1×105 and about 1 ×107 of the presently disclosed cells comprising the GPC3-targeted CAR. In certain embodiments, the presently disclosed composition comprises between about 1×106 and about 1 ×108 of the presently disclosed cells comprising the GPC3-targeted CAR. In certain embodiments, the presently disclosed composition comprises between about 1×106 and about 5×108 of the presently disclosed cells comprising the GPC3-targeted CAR. More effective cells may be administered in even smaller numbers. Usually, at least about 1 × 105 cells will be administered, eventually reaching about 1 × 1010 or more. In certain embodiments, the presently disclosed composition comprises at least about 1×105, 5×105, 1×106, about 5×106, about 1×107, about 2.5×107, about 5×107, about 1×108, about 1.5×108, about 2×108, or about 5×108 of the presently disclosed cells comprising the GPC3-targeted CAR. In certain embodiments, the presently disclosed composition comprises about 1×106 of the presently disclosed cells comprising the GPC3-targeted CAR. The precise determination of what would be considered an effective dose can be based on factors individual to each subject, including their size, age, sex, weight, and condition of the particular subject. Dosages can be readily ascertained by those skilled in the art from this disclosure and the knowledge in the art. The compositions can be administered multiple times at these dosages. [00776] The presently disclosed CAR-modified cells may be administered either alone, or as a pharmaceutical composition in combination with diluents and/or with other components such as IL- 2, IL-15, or other cytokines or cell populations. Briefly, pharmaceutical compositions may comprise NAI-1540294631v3 224 a target cell population as described herein, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. Such compositions may comprise buffers such as neutral buffered saline, phosphate buffered saline and the like; carbohydrates such as glucose, mannose, sucrose or dextrans, mannitol; proteins; polypeptides or amino acids such as glycine; antioxidants; chelating agents such as EDTA or glutathione; adjuvants (e.g., aluminum hydroxide); and preservatives. Compositions for use in the disclosed methods are in some embodiments formulated for intravenous administration. The quantity and frequency of administration will be determined by such factors as the condition of the patient, and the severity of the patient's disease, although appropriate dosages may be determined by clinical trials. [00777] The administration of the disclosed compositions may be carried out in any convenient manner, including by injection, transfusion, or implantation. The presently disclosed compositions can be administered by any method known in the art including, but not limited to, intravenous administration, intradermal administration, subcutaneous administration, intranodal administration, intratumoral administration, intramedullary administration, intramuscular administration, intrathecal administration, intrapleural administration, intraosseous administration, intraperitoneal administration, pleural administration, and direct administration to the subject. In certain embodiments, the composition is administered to a subject intravenously. The compositions can be injected directly into a tumor, lymph node, or site of disease. 5.9. Methods of Treatment [00778] The presently disclosed GPC3-targeted CAR-expressing cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used for treating a GPC3-associated disease, disorder, or condition, including one or more symptoms of the disease, disorder, or condition. Thus, the presently disclosed subject matter provides methods of treating a GPC3-associated disease, disorder, or condition. In certain embodiments, the method comprises administering to a subject in need thereof a presently disclosed anti-GPC3 antibody, antigen-binding fragment thereof, multispecific molecule, conjugate, or composition. In certain embodiments, the method comprises administering to a subject in need thereof a composition comprising cells comprising a presently disclosed GPC3-targeted CAR. In certain embodiments, the subject suffers from or is diagnosed with a GPC3-associated disease, disorder, or condition. In certain embodiments, the subject is human. NAI-1540294631v3 225 [00779] Immune effector cells expressing the presently disclosed GPC3-targeted CARs can elicit an immune response against GPC3-expressing cells. The immune response elicited by the disclosed CAR-modified immune effector cells may be an active or a passive immune response. In addition, the CAR-mediated immune response may be part of an adoptive immunotherapy approach in which CAR-modified immune effector cells induce an immune response specific to GPC3. [00780] Adoptive transfer of immune effector cells expressing CARs is a promising anti-cancer therapeutic. Following the collection of a subject’s immune effector cells, the cells may be genetically engineered to express the presently disclosed GPC3-targeted CARs, then infused back into the subject. Moreover, cells (e.g., immune effector cells) obtained from a donor other than the subject (e.g., allogeneic to the subject) may be genetically engineered to express the disclosed GPC3-targeted CARs, then the CAR-expressing cells infused into the subject. In certain embodiments, the cells comprising a GPC3-targeted CAR polypeptide are allogeneic EBV-specific cytotoxic T cells. [00781] The presently disclosed subject matter provides various methods of using the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions. The presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions can be used in a therapy or medicament. For example, the presently disclosed subject matter provides methods for inducing and/or increasing an immune response in a subject in need thereof. The presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions can be used for treating a GPC3-associated disease or disorder in a subject. In certain embodiments, the GPC3-associated disease or disorder in a subject is a tumor. Thus, the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions can be used for reducing tumor burden in a subject, and/or prolonging the survival of a subject suffering from a tumor. The presently disclosed cells and compositions comprising thereof can reduce the number of tumor cells, reduce tumor size, and/or eradicate the tumor in the subject. In certain embodiments, each of the above-noted method comprises administering the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates, or compositions to achieve the desired effect, e.g., palliation of an existing condition or prevention of recurrence. For treatment, the amount administered is an amount effective in producing the desired effect. An effective amount can be provided in one or a series of administrations. An effective amount can be provided in a bolus or by continuous perfusion. NAI-1540294631v3 226 [00782] In certain embodiments, the GPC3-associated disease or disorder overexpresses GPC3. Non-limiting examples of GPC3-associated disease or disorder include hepatocellular carcinoma, hepatoblastoma, lung squamous cell carcinoma, ovarian yolk sac tumor, melanoma, and urothelial carcinoma. In certain embodiments, the GPC3-associated disease or disorder is a tumor. In certain embodiments, the GPC3-associated disease or disorder is cancer. In certain embodiments, the GPC3-associated disease or disorder is hepatocellular carcinoma. [00783] In certain embodiments, the subject is a human subject. The subjects can have an advanced form of disease, in which case the treatment objective can include mitigation or reversal of disease progression, and/or amelioration of side effects. The subjects can have a history of the condition, for which they have already been treated, in which case the therapeutic objective will typically include a decrease or delay in the risk of recurrence. [00784] As a consequence of surface expression of a presently disclosed GPC3-targeted CAR, adoptively transferred cells are endowed with augmented and selective cytolytic activity at the tumor site. Furthermore, subsequent to their localization to tumor and their proliferation, the cells turn the tumor site into a highly conductive environment for a wide range of cells involved in the physiological anti-tumor response. [00785] In certain embodiments, the presently disclosed cells, anti-GPC3 antibodies, antigen- binding fragments thereof, multispecific molecules, conjugates and compositions are administered to a subject in conjunction with (e.g., before, simultaneously or following) any number of relevant treatment modalities. In certain embodiments, the presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used in combination with chemotherapy, radiation, immunosuppressive agents, immunoablative agents, cytoxin, and cytokines. [00786] In certain embodiments, the GPC3-associated disease or disorder is cancer. In certain embodiments, the cancer can be any neoplasm or tumor for which radiotherapy is currently used. Alternatively, the cancer can be a neoplasm or tumor that is not sufficiently sensitive to radiotherapy using standard methods. A representative but non-limiting list of cancers that the disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used to treat include hepatocellular carcinoma, hepatoblastoma, lung squamous cell carcinoma, ovarian yolk sac tumor, melanoma, and urothelial carcinoma. [00787] The presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used in combination with any compound, moiety or group which has a cytotoxic or cytostatic effect. Drug moieties include NAI-1540294631v3 227 chemotherapeutic agents, which may function as microtubulin inhibitors, mitosis inhibitors, topoisomerase inhibitors, or DNA intercalators, and particularly those which are used for cancer therapy. [00788] The presently disclosed cells, anti-GPC3 antibodies, antigen-binding fragments thereof, multispecific molecules, conjugates and compositions can be used in combination with a checkpoint inhibitor. The two known inhibitory checkpoint pathways involve signaling through the cytotoxic T- lymphocyte antigen-4 (CTLA-4) and programmed-death 1 (PD-1) receptors. These proteins are members of the CD28-B7 family of co-signaling molecules that play important roles throughout all stages of T cell function. The PD-1 receptor (also known as CD279) is expressed on the surface of activated T cells. Its ligands, PD-L1 (B7-H1; CD274) and PD-L2 (B7-DC; CD273), are expressed on the surface of APCs such as dendritic cells or macrophages. PD-L1 is the predominant ligand, while PD-L2 has a much more restricted expression pattern. When the ligands bind to PD-1, an inhibitory signal is transmitted into the T cell, which reduces cytokine production and suppresses T- cell proliferation. Checkpoint inhibitors include, but are not limited to antibodies that block PD-1 (e.g., Nivolumab (BMS-936558 or MDX1106), CT-011, MK-3475), PD-L1 (e.g., MDX-1105 (BMS-936559), MPDL3280A, MSB0010718C), PD-L2 (e.g., rHIgM12B7), CTLA-4 (e.g., Ipilimumab (MDX-010), Tremelimumab (e.g., CP-675,206)), IDO, B7-H3 (e.g., MGA271), B7-H4, TIM3, LAG-3 (e.g., BMS-986016). Techniques for combining CARs with checkpoint inhibitors in immune effector cells and use thereof for the treatment of various disorders are described, for example, in WO 2017/040945, which is incorporated by reference herein. [00789] T cell receptor activation plus co-stimulation can help generate optimal “killer” CD8 T cell responses, which can be provided through ligation of tumor necrosis factor receptor family members, including OX40 (CD134) and 4-1BB (CD137). OX40 is of particular interest as treatment with an activating (agonist) anti-OX40 mAb augments T cell differentiation and cytolytic function leading to enhanced anti-tumor immunity against a variety of tumors. [00790] In certain embodiments, the presently disclosed method comprises administering to the subject a second therapeutic agent. In certain embodiments, the second therapeutic agent is an antimetabolite, such as methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, fludarabine, 5- fluorouracil, decarbazine, hydroxyurea, asparaginase, gemcitabine or cladribine. [00791] In certain embodiments, the second therapeutic agent is an alkylating agent, such as mechlorethamine, thioepa, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, dacarbazine (DTIC), procarbazine, mitomycin C, cisplatin and other platinum derivatives, such as carboplatin. NAI-1540294631v3 228 [00792] In certain embodiments, the second therapeutic agent is an anti-mitotic agent, such as taxanes, for instance docetaxel, and paclitaxel, and vinca alkaloids, for instance vindesine, vincristine, vinblastine, and vinorelbine. [00793] In certain embodiments, the second therapeutic agent is a topoisomerase inhibitor, such as topotecan or irinotecan, or a cytostatic drug, such as etoposide and teniposide. [00794] In certain embodiments, the second therapeutic agent is a growth factor inhibitor, such as an inhibitor of ErbBl (EGFR) (such as an EGFR antibody, e.g. zalutumumab, cetuximab, panitumumab or nimotuzumab or other EGFR inhibitors, such as gefitinib or erlotinib), another inhibitor of ErbB2 (HER2/neu) (such as a HER2 antibody, e.g. trastuzumab, trastuzumab-DM l or pertuzumab) or an inhibitor of both EGFR and HER2, such as lapatinib). [00795] In certain embodiments, the second therapeutic agent is a tyrosine kinase inhibitor, such as imatinib (Glivec, Gleevec STI571) or lapatinib. [00796] In certain embodiments, the second therapeutic agent is a cytokine, growth factor, chemokine, or a combination thereof. Non-limiting examples of cytokines and growth factors include IFNγ, IL-2, IL-4, IL-6, IL-7, IL-10, IL-12, IL-13, IL-15, IL-18, IL-23, IL-24, IL-27, IL-28a, IL-28b, IL-29, KGF, IFNα (e.g., INFα2b), IFN , GM-CSF, CD40L, Flt3 ligand, stem cell factor, ancestim, and TNFα. Non-limiting examples of chemokines include Glu-Leu-Arg (ELR)-negative chemokines such as IP-10, MCP-3, MIG, and SDF-la from the human CXC and C-C chemokine families. Suitable cytokines include cytokine derivatives, cytokine variants, cytokine fragments, and cytokine fusion proteins. [00797] In certain embodiments, the second therapeutic agent is a cell cycle control/apoptosis regulator (or “regulating agent”). A cell cycle control/apoptosis regulator may include molecules that target and modulate cell cycle control/apoptosis regulators such as (i) cdc-25 (such as NSC 663284), (ii) cyclin-dependent kinases that overstimulate the cell cycle (such as flavopiridol (L868275, HMR1275), 7-hydroxystaurosporine (UCN-01, KW-2401), and roscovitine (R- roscovitine, CYC202)), and (iii) telomerase modulators (such as BIBR1532, SOT-095, GRN163 and compositions described in for instance US 6,440,735 and US 6,713,055) . Non-limiting examples of molecules that interfere with apoptotic pathways include TNF-related apoptosis-inducing ligand (TRAIL)/apoptosis-2 ligand (Apo-2L), antibodies that activate TRAIL receptors, IFNs, and anti- sense Bcl-2. [00798] In certain embodiments, the second therapeutic agent is a hormonal regulating agent, such as agents useful for anti-androgen and anti-estrogen therapy. Examples of such hormonal regulating agents are tamoxifen, idoxifene, fulvestrant, droloxifene, toremifene, raloxifene, NAI-1540294631v3 229 diethylstilbestrol, ethinyl estradiol/estinyl, an antiandrogene (such as flutaminde/eulexin), a progestin (such as such as hydroxyprogesterone caproate, medroxy- progesterone/provera, megestrol acepate/megace), an adrenocorticosteroid (such as hydrocortisone, prednisone), luteinizing hormone- releasing hormone (and analogs thereof and other LHRH agonists such as buserelin and goserelin), an aromatase inhibitor (such as anastrazole/arimidex, aminoglutethimide/cytraden, exemestane) or a hormone inhibitor (such as octreotide/sandostatin). [00799] In certain embodiments, the second therapeutic agent is an anti-cancer nucleic acid or an anti-cancer inhibitory RNA molecule. [00800] Combined administration, as described above, may be simultaneous, separate, or sequential. For simultaneous administration the agents may be administered as one composition or as separate compositions, as appropriate. [00801] In certain embodiments, the presently disclosed cells, anti-GPC3 antibodies, antigen- binding fragments thereof, multispecific molecules, conjugates and compositions are administered in combination with radiotherapy. Radiotherapy may comprise radiation or associated administration of radiopharmaceuticals to a subject is provided. The source of radiation may be either external or internal to the patient being treated (radiation treatment may, for example, be in the form of external beam radiation therapy (EBRT) or brachytherapy (BT)). Radioactive elements that may be used in practicing such methods include, e.g., radium, cesium-137, iridium-192, americium-241, gold-198, cobalt-57, copper-67, technetium-99, iodide-123, iodide-131, and indium-111. [00802] In certain embodiments, the presently disclosed cells, anti-GPC3 antibodies, antigen- binding fragments thereof, multispecific molecules, conjugates and compositions are administered in combination with surgery. [00803] The GPC3-targeted CAR-T cells may be designed in several ways that enhance tumor cytotoxicity and specificity, evade tumor immunosuppression, avoid host rejection, and prolong their therapeutic half-life. TRUCK (T-cells Redirected for Universal Cytokine Killing) T cells for example, possess a CAR but are also engineered to release cytokines such as IL-12 that promote tumor killing. Because these cells are designed to release a molecular payload upon activation of the CAR once localized to the tumor environment, these CAR-T cells are sometimes also referred to as ‘armored CARs’. Several cytokines as cancer therapies are being investigated both pre-clinically and clinically, and may also prove useful when similarly incorporated into a TRUCK form of CAR- T therapy. Among these include IL-2, IL-3. IL-4, IL-5, IL-6, IL-7, IL-10, IL-12, IL-13, IL-15, IL- 18, M-CSF, GM-CSF, IFN-α, IFN-γ, TNF-α, TRAIL, FLT3 ligand, Lymphotactin, and TGF-β (Dranoff 2004). “Self-driving” or “homing” CAR-T cells are engineered to express a chemokine NAI-1540294631v3 230 receptor in addition to their CAR. As certain chemokines can be upregulated in tumors, incorporation of a chemokine receptor aids in tumor trafficking to and infiltration by the adoptive T- cell, thereby enhancing both specificity and functionality of the CAR-T (Moon 2011). Universal CAR-T cells also possess a CAR, but are engineered such that they do not express endogenous TCR (T-cell receptor) or MHC (major histocompatibility complex) proteins. Removal of these two proteins from the signaling repertoire of the adoptive T-cell therapy prevents graft-versus-host- disease and rejection, respectively. Armored CAR-T cells are additionally so named for their ability to evade tumor immunosuppression and tumor-induced CAR-T hypofunction. These particular CAR-Ts possess a CAR, and may be engineered to not express checkpoint inhibitors. Alternatively, these CAR-Ts can be co-administered with a monoclonal antibody (mAb) that blocks checkpoint signaling. Administration of an anti-PD-L1 antibody significantly restored the killing ability of CAR TILs (tumor infiltrating lymphocytes). While PD1-PDL1 and CTLA-4-CD80/CD86 signaling pathways have been investigated, it is possible to target other immune checkpoint signaling molecules in the design of an armored CAR-T including LAG-3, Tim-3, IDO-1, 2B4, and KIR. Other intracellular inhibitors of TILs include phosphatases (SHP1), ubiquitin-ligases (i.e., cbl-b), and kinases (i.e., diacylglycerol kinase) . Armored CAR-Ts may also be engineered to express proteins or receptors that protect them against or make them resistant to the effects of tumor-secreted cytokines. For example, CTLs (cytotoxic T lymphocytes) transduced with the double negative form of the TGF-β receptor are resistant to the immunosuppression by lymphoma secreted TGF-β. These transduced cells showed notably increased antitumor activity in vivo when compared to their control counterparts. [00804] Tandem and dual CAR-T cells are unique in that they possess two distinct antigen binding domains. A tandem CAR contains two sequential antigen binding domains facing the extracellular environment connected to the intracellular costimulatory and stimulatory domains. A dual CAR is engineered such that one extracellular antigen binding domain is connected to the intracellular costimulatory domain and a second, distinct extracellular antigen binding domain is connected to the intracellular stimulatory domain. Because the stimulatory and costimulatory domains are split between two separate antigen binding domains, dual CARs are also referred to as “split CARs”. In both tandem and dual CAR designs, binding of both antigen binding domains is necessary to allow signaling of the CAR circuit in the T-cell. Because these two CAR designs have binding affinities for different, distinct antigens, they are also referred to as “bi-specific” CARs. [00805] To better control CAR-T therapy and prevent against unwanted side effects, a variety of features have been engineered including off-switches, safety mechanisms, and conditional control NAI-1540294631v3 231 mechanisms. Both self-destruct and marked/tagged CAR-T cells for example, are engineered to have an “off-switch” that promotes clearance of the CAR-expressing T-cell. A self-destruct CAR-T contains a CAR, but is also engineered to express a pro-apoptotic suicide gene or “elimination gene” inducible upon administration of an exogenous molecule. A variety of suicide genes may be employed for this purpose, including HSV-TK (herpes simplex virus thymidine kinase), Fas, iCasp9 (inducible caspase 9), CD20, MYC TAG, and truncated epidermal growth factor receptor (EGFRt) polypeptide (endothelial growth factor receptor). HSK for example, converts the prodrug ganciclovir (GCV) into GCV-triphosphate that incorporates itself into replicating DNA, ultimately leading to cell death. iCasp9 is a chimeric protein containing components of FK506-binding protein that binds the small molecule AP1903, leading to caspase 9 dimerization and apoptosis. In certain embodiments, the suicide gene is an EGFRt polypeptide. The EGFRt polypeptide can enable T-cell elimination by administering anti-EGFR monoclonal antibody (e.g., cetuximab). EGFRt can be covalently joined to the upstream of the GPC3-targeted CAR. The suicide gene can be included within the vector comprising nucleic acids encoding a presently disclosed GPC3-targeted CAR. In this way, administration of a prodrug designed to activate the suicide gene (e.g., a prodrug (e.g., AP1903 that can activate iCasp-9) during malignant T-cell transformation (e.g., GVHD) triggers apoptosis in the suicide gene-activated cells expressing the GPC3-targeted CAR. The incorporation of a suicide gene into the a presently disclosed GPC3-targeted CAR gives an added level of safety with the ability to eliminate the majority of receptor-expressing cells within a very short time period. A presently disclosed cell incorporated with a suicide gene can be pre-emptively eliminated at a given timepoint post the cell infusion, or eradicated at the earliest signs of toxicity. [00806] A marked/ tagged CAR-T cell however, is one that possesses a CAR but also is engineered to express a selection marker. Administration of a mAb against this selection marker may promote clearance of the CAR-T cell. Truncated EGFR is one such targetable antigen by the anti-EGFR mAb, and administration of cetuximab works to promotes elimination of the CAR-T cell. CARs created to have these features are also referred to as sCARs for “switchable CARs”, and RCARs for ‘regulatable CARs’. A “safety CAR”, also known as an “inhibitory CAR” (iCAR), is engineered to express two antigen binding domains. One of these extracellular domains is directed against a tumor related antigen and bound to an intracellular costimulatory and stimulatory domain. The second extracellular antigen binding domain however is specific for normal tissue and bound to an intracellular checkpoint domain such as CTLA-4, PD-1, or CD45. Incorporation of multiple intracellular inhibitory domains to the iCAR is also possible. Some inhibitory molecules that may provide these inhibitory domains include B7-H1, B7-1, CD160, PIH, 2B4, CEACAM (CEACAM-1. NAI-1540294631v3 232 CEACAM-3, and/or CEACAM-5), LAG-3, TIGIT, BTLA, LAIR1, and TGFβ-R. In the presence of normal tissue, stimulation of this second antigen binding domain can inhibit the CAR. It should be noted that due to this dual antigen specificity, iCARs are also a form of bi-specific CAR-T cells. The safety CAR-T engineering enhances specificity of the CAR-T cell for tumor tissue, and is advantageous in situations where certain normal tissues may express very low levels of a tumor associated antigen that would lead to off target effects with a standard CAR (Morgan 2010). A conditional CAR-T cell expresses an extracellular antigen binding domain connected to an intracellular costimulatory domain and a separate, intracellular costimulator. The costimulatory and stimulatory domain sequences are engineered in such a way that upon administration of an exogenous molecule the resultant proteins will come together intracellularly to complete the CAR circuit. In this way, CAR-T activation can be modulated, and possibly even “fine-tuned” or personalized to a specific patient. Similar to a dual CAR design, the stimulatory and costimulatory domains are physically separated when inactive in the conditional CAR; for this reason these too are also referred to as a “split CAR”. [00807] In certain embodiments, two or more of these engineered features may be combined to create an enhanced, multifunctional CAR-T. For example, it is possible to create a CAR-T cell with either dual- or conditional-CAR design that also releases cytokines like a TRUCK. In certain embodiments, a dual-conditional CAR-T cell could be made such that it expresses two CARs with two separate antigen binding domains against two distinct cancer antigens, each bound to their respective costimulatory domains. The costimulatory domain would only become functional with the stimulatory domain after the activating molecule is administered. For this CAR-T cell to be effective the cancer must express both cancer antigens and the activating molecule must be administered to the patient; this design thereby incorporating features of both dual and conditional CAR T cells. 6. EXAMPLE [00808] The practice of the presently disclosed subject matter employs, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are well within the purview of the skilled artisan. [00809] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the presently disclosed cells and compositions, and are not intended to limit the scope of what the inventors regard as their invention. Example 1. Generation of GPC3-Targeted scFv-Fc Fusion Proteins NAI-1540294631v3 233 [00810] A total of 16 human binders for human GPC3 were obtained as described in Section 5.3.1. To characterize the binders, 16 binders were formatted as fusion proteins with IgG1 (Kappa) Fc domain and produced. [00811] Briefly, the VH and VL of a binder were coupled through a linker (e.g., a linker consisting of the amino acid sequence set forth in SEQ ID NO: 1) to form a single chain variable region (scFv). The scFv was further fused with a C-terminal Fc domain (e.g., a Fc domain comprising the amino acid sequence set forth in SEQ ID NO: 357) in order to purify the expressed scFv. The fusion protein was designated as “scFv-Fc.” The 16 scFv-Fc fusion proteins were designated as “ATA001 VH-VK Fc” (or “1-VH-linker-VL” shown in Figures 1A-1E), “ATA002 VH-VK Fc” (or “2-VH- linker-VL” shown in Figures 1A-1E), “ATA003 VH-VK Fc” (or “3-VH-linker-VL” shown in Figures 1A-1E), “ATA004 VH-VK Fc” (or “4-VH-linker-VL” shown in Figures 1A-1E), “ATA005 VH-VK Fc” (or “5-VH-linker-VL” shown in Figures 1A-1E), “ATA006 VH-VK Fc” (or “6-VH- linker-VL” shown in Figures 1A-1E), “ATA007 VH-VK Fc” (or “7-VH-linker-VL” shown in Figures 1A-1E), “ATA008 VH-VK Fc” (or “8-VH-linker-VL” shown in Figures 1A-1E), “ATA009 VH-VK Fc” (or “9-VH-linker-VL” shown in Figures 1A-1E), “ATA010 VH-VK Fc” (or “10-VH- linker-VL” shown in Figures 1A-1E), “ATA011 VH-VK Fc” (or “11-VH-linker-VL” shown in Figures 1A-1E), “ATA012 VH-VK Fc” (or “12-VH-linker-VL” shown in Figures 1A-1E), “ATA013 VH-VK Fc” (or “13-VH-linker-VL” shown in Figures 1A-1E), “ATA014 VH-VK Fc” (or “14-VH- linker-VL” shown in Figures 1A-1E), “ATA015 VH-VK Fc” (or “15-VH-linker-VL” shown in Figures 1A-1E), and “ATA016 VH-VK Fc” (or “16-VH-linker-VL” shown in Figures 1A-1E). To facilitate expression, codon optimization for scFv-Fc was performed as described in Section 5.3.1. [00812] To produce scFv-Fc of 16 binders, plasmids encoding scFv-Fc were introduced into Expi293F cells or ExpiCHO-S cells by transient transfection using standard procedures. The scFv- Fc of 16 binders were purified from culture supernatant produced using standard procedures of Protein A magnetic bead affinity purification. The scFv-Fc fusion proteins for 16 binders were expressed and purified with a purity of greater than 90%. Example 2. Binding Activities of scFv-Fc Fusion Proteins [00813] The binding activities of the scFv-Fc fusion proteins described in Example 1 to human GPC3 were assessed by using enzyme-linked immunosorbent assay (ELISA) and fluorescence- activated cell sorting (FACS). [00814] As shown in the ELISA results of Table 17, all tested scFv-Fc fusion proteins showed specific binding to GPC36×His protein. In addition, the FACS results demonstrate the specific binding of all tested scFv-Fc fusion proteins to human GPC3. For example, as shown in Figures 1A NAI-1540294631v3 234 to 1E, all tested scFv-Fc fusion proteins bound to the positive cell line CHO-K1/human GPC3 clone 9(+) cells expressing human GPC3, but not to the negative cell line CHO-K1(-). The GPC3-targeted GC33 and Y035 scFvs were used as positive controls. The EC50 (ng/ml) of GC33 was 9.578. Y035 affinity values were 3.96 × 105 for Ka (1/Ms), 2.51 × 10-5 for Kd (1/s), and 6.34 × 10-11 for KD (M), obtained from the literature. Table 17. ELISA Results of Purified scFv-Fc Proteins Binder EC50 (ng/ml) ATA001 VH-VK Fc 12.140 Example 3. Epitope Binn
Figure imgf000237_0001
[00815] The scFv-Fc fusion proteins described in Example 1 were tested for competitive binding to human GPC3 by competition ELISA. [00816] In competition ELISA, each of biotinylated scFv-Fv fusion proteins at optimal concentration cross-incubated with unlabeled scFv-Fc fusion proteins in 96-well plate for competitive binding to immobilized human GPC3. After washing, binding of biotinylated scFv-Fc protein to GPC3 was detected by streptavidin-HRP secondary antibody. GC33 antibody against GPC3 was used as benchmark. Competitive binding activity was used to cluster the binders by epitope bins. [00817] Final concentration of each biotinylated scFv-Fc protein designated as “-Bio” used for competition ELISA were shown in Table 18. Self-competition was highlighted. Results indicated that the binders exhibited various levels of competition. Based on the competitive and non- competitive relationship, the binders were grouped by epitope bins. The 16 binders were shown to bind to five distinct epitopes: Epitope 1, Epitope 2, Epitope 3, Epitope 4, and Epitope 5. For example, ATA001, ATA002, ATA003, ATA004, ATA005, ATA006, and ATA011 can be binned together and in the same group as GC33 antibody that bound to Epitope 1. ATA007, ATA009, and NAI-1540294631v3 235 ATA015 were shown to bind to Epitope 2. ATA008, ATA009, ATA010, and ATA013 were shown to bind to Epitope 3; ATA014 and ATA016 were shown to bind to Epitope 4; and ATA012 was shown to bind to Epitope 5. Epitope 1 is within amino acids 524-563 of human GPC3 (e.g., one comprising the amino acid sequence set forth in SEQ ID NO: 22). Epitope 3 is at the C-terminus of GPC3. NAI-1540294631v3 236 - 61 c 0 F l m AKo i / 1 0 0 7 2 1 9 3 4 9 0 7 7 5 1 4 1 7 7 6 8 5 5 3 3 9 T V - Bg n 0 3 . 1 4 . 1 2 . 1 3 . 1 . 1 . 1 . 1 . 1 . 8 . 1 . 4 . 0 . AH . 4 1 1 1 1 1 1 0 1 0 0
Figure imgf000239_0001
A T V- B - 0 . 7 . 7 . 7 . . . . . . . . . . . AH 0 1 1 1 0 1 0 1 0 0 1 1 1 1 V 0 5 - 40 c 0 F l m 5 9 1 0 3 4 5 7 0 2 6 AKo i / T V - Bg n 8 5 3 . 6 1 0 . 9 0 0 . 6 0 3 . 3 1 . 7 1 . 3 3 . 2 1 . 8 0 . 8 0 . 7 2 0 . 4 5 9 . 2 2 . AH . 7 1 1 1 0 0 0 1 1 0 1 V kr o a i m B h 3 - l 3 ym c C d / g 1 2 0 6 1 4 1 8 1 5 9 1 1 6 4 5 3 0 9 6 7 8 8 8 8 1 ne Go b n i t 0 . 7 . 1 0 . 0 2 . 0 6 . 1 6 . 1 6 . 1 9 . 0 4 . 0 2 . 0 6 . 1 6 . 1 6 . 1 7 . 1 B n 3 a )n 3 l c F c F c c c c F c F c F c c c b oi At 3 a m F F F F F F r C / g K V K V K K K K V K V K V V V V K V K K g t ni n l t e e c o G rt k u r 0 - l a 2 Hm / - l Hm / - l Hm / - l Hm / - l Hm - l - l - l / Hm / Hm / Hm / - l V Hm / - l V Hm / - l Hm / p n mo n V o c o l c mh y g d 4 0 u Vg 0 7 a c 0 u Vg 0 8 0 u V g 0 9 0 u Vg 0 3 0 u Vg 0 5 0 u V 0 6 0 u g V 0 2 1 u g V g 0 3 1 u Vg 0 4 u V g 0 6 u 0 n o b it 0 2 2 2 2 5 A 0 0 0 0 0 5 A 0 5 5 5 1 5 1 5 A 0 A 0 0 0 C n i e n T A T A T A T A T T T A A A F B a A T A T T T ( A A A A A A A A A Example 4. Generation of GPC3-targeted CAR T Cells [00818] Twelve (12) GPC3-targeted CARs were constructed. In particular, six (6) GPC3 binders ATA001, ATA002, ATA004, ATA008, ATA009, and ATA016 in both VH-VL orientation and VL-VH orientation were used to generate SFG 28z1XX P2A LNGFR CAR constructs. These twelve CARs were designated as follows: “1_28z1XX (vH-vK)” (also referred to as “ATA017 VH-VK CAR”), “1_28z1XX (vK-vH)” (also referred to as “ATA017 VK-VH CAR”), “2_28z1XX (vH-vK)” (also referred to as “ATA018 VH-VK CAR”), “2_28z1XX (vK- vH)” (also referred to as “ATA018 VK-VH CAR”), “4_28z1XX (vH-vK)” (also referred to as “ATA019 VH-VK CAR”), “4_28z1XX (vK-vH)” (also referred to as “ATA019 VK-VH CAR”), “8_28z1XX (vH-vK)” (also referred to as “ATA020 VH-VK CAR”), “8_28z1XX (vK-vH)” (also referred to as “ATA020 VK-VH CAR”), “9_28z1XX (vH-vK)” (also referred to as “ATA021 VH-VK CAR”), “9_28z1XX (vK-vH)” (also referred to as “ATA021 VK-VH CAR”), “16_28z1XX (vH-vK)” (also referred to as “ATA022 VH-VK CAR”), and “16_28z1XX (vK- vH)” (also referred to as “ATA022 VK-VH CAR). The general structure of the CAR is illustrated in Figure 2A. As shown in Figure 2A, the scFv was preceded by a leader peptide and followed by CD28 hinge-transmembrane-intracellular regions, and ITAM-mutated CD3ζ intracellular domains (“1XX”) linked to a P2A sequence to induce co-expression of truncated low-affinity nerve growth factor receptor (LNGFR). 48-hour after initiating T cell activation, T cells were transduced with retroviral supernatants by centrifugation. Transduction efficiencies were determined 4 days later by flow cytometry. As shown in Figure 2B, the flow cytometric analysis shows the expression levels of LNGFR for the indicated constructs in the VH-VK and VK-VH orientation. Untransduced T (UT) cells were used as a negative control. The results indicate that the twelve (12) GPC3-targeted CARs were successfully transduced into and expressed by the host T cells and showed comparable levels of transduction efficiency. [00819] The T cell exhaustion of the twelve (12) GPC3-targeted CARs was assessed by CD3/CD28 beads activation. On day 8 post activation, a fluorescence-activated cell sorting (FACS) analysis was performed to analyze the exhaustion markers LAG3, PD1, and TIM3 on the CAR T cells. Analysis shows no difference in expression of LAG3, PD1 or TIM3 between VH-VK and VK-VH orientation CAR T cells (data not shown). The results indicate that all 6 GPC3 scFvs have similar or lower levels of exhaustion markers expressed compared to GC33 or Y035 CARs. 238 NAI-1540294631v3 Example 5. In Vitro Cytotoxicity Activities of GPC3-Targeted CAR T Cells [00820] The in vitro cytotoxicity of the GPC3-targeted CAR T cells described in Example 4 against cells expressing different levels of GPC3 was assessed by Cytotoxic T Lymphocyte (CTL) assays. [00821] Nalm-6 GPC3+ (NALM-6 target cell has been engineered to express GPC3), Huh7, and PLC/PRF/5-expressing FFLuc-GFP were served as target cells. The GPC3 expression levels of Nalm-6 GPC3+, Huh7, and PLC/PRF/5 tumor cells are shown in Figure 3. Nalm-6 GPC3+ cells expressed ~5600 GPC3 molecules, Huh-7 cells expressed ~3000 GPC3 molecules, and PLC/PRF/5 cells expressed ~500 GPC3 molecules. Tumor cell killing was measured in a bioluminescence (BLI) assay at the indicated effector:target (E:T) ratios. The effector and tumor target cells were co-cultured in triplicates at the indicated effector/target ratio using black-walled 96-well plates with 1×104 target cells in a total volume of 100 µ per well in medium. Target cells alone were plated at the same cell density to determine the maximal luciferase expression (relative light units (RLU); 18 hours later, 100 µL luciferase substrate was directly added to each well. Emitted light was detected in a luminescence plate reader. Lysis was determined as (1 – (RLUsample)/(RLUmax)) × 100. [00822] As shown in Figure 4A, all tested presently disclosed GPC3 binders showed target- specific killing of the Nalm6 GPC3+ cells, where the killing was quantitatively similar to the 1928z 1XX CAR T.1928z 1XX was used as a positive control in this assay as it is an anti-CD19 CAR T that functions to kill Nalm-6 wildtype (WT) cells because Nalm-6 WT cells naturally express CD19 on their surface. As shown in Figure 4B, the tested presently disclosed GPC3- targeted CARs showed very low levels of non-specific killing of Nalm-6 WT tumor cells, with certainly much lower quantitatively than the levels of cell killing shown in Figure 4A. These combined data, therefore, demonstrate that all of the tested presently disclosed GPC3-targeted CARs effectively kill target cells in a GPC3-specific manner. [00823] In addition, all tested presently disclosed GPC3 binders showed target-specific killing of the Huh-7 tumor cells. See Figure 5. Similarly, as shown in Figure 6, all tested presently disclosed GPC3 binders showed target-specific killing of the PLC/PRF/5 tumor cells that had a low GPC3 expression level on the cell surface. In addition, all tested presently disclosed GPC3 binders showed improved cytotoxicity as compared to Y035 and GC33 CAR T-cells. NAI-1540294631v3 239 [00824] In summary, all tested presently disclosed GPC3 binders showed in vitro anti-tumor activity against GPC3-expressing tumor cells. Epitope 1, 3 and 4 on GPC3 were capable of activating CAR T-cells leading to cytotoxicity against tumor cells. All GPC3 binders had improved or comparable cytotoxicity against Nalm-6 GPC3+ and Huh-7 tumor cells as compared to GC33 CAR T-cells. Binders 1, 4, 8, 9 and 16 showed superior cytotoxicity against PLC/PRF/5 tumor cells as compared to GC33_28z1XX CAR T-cells. In addition, the results show that the orientation of the VH and VK (or VL) chains within the scFv does not seem to have a significant impact on the efficacy of the CAR composed of the scFvs, e.g., both the VL- VH and VH-VL orientations within the scFvs produced functional CARs possessing anti-tumor activities. Therefore, moving forward, VH-VK orientation CAR T cells were used. Example 6. In Vivo Antitumor Efficacy and Toxicity of GPC3-Targeted CAR T Cells [00825] To evaluate the in vivo antitumor efficacy and toxicity of the GPC3-targeted CAR T cells described in Example 4, 6- to 8-week-old NOD/SCID/IL-2Rg-null (NSG) female mice (n=5) were injected with 5 × 106 mCherry-luciferase overexpressing Huh-7D12 cells subcutaneously on day 0 followed by intravenous (i.v.) injection of 5 × 106 CAR-positive T cells on day 9. Tumor volume (length and width) was measured 3 times per week with caliper and calculated as volume = 0.5 × length × width2. Body weight was measured 3 times a week to assess the acute and severe toxicities associated with the treatment. All animals were sacrificed according to IACUC protocol standards. An experimental layout was illustrated in Figure 7. [00826] 12 days post CAR-T injection, all mice from CAR-T groups with binder 1 (also referred to as “GPC3-1”), binder 2 (also referred to as “GPC3-2”) and binder 4 (also referred to as “GPC3-4”), together with 3 mice from CAR-T group with binder 9 (also referred to as “GPC3-9”) died from toxicity, with symptoms including significant body weight drop, ruffle and hunch. See Figure 8. Thus, these four binder candidates were eliminated from screening due to safety concerns. [00827] Conversely, CAR-T groups with binder 8 (also referred to as “8_28z1XX (vH-vK)”) and binder 16 (also referred to as “16_28z1XX (vH-vK)”) efficiently reduced tumor size over time and prolonged the overall survival of tumor bearing mice. All mice in CAR-T group with binder 8 achieved tumor free and sustained until termination of the study, while 3 of the 5 mice from binder 16 group achieved tumor free. See Figure 9. 30 days post CAR-T injection, mice in CAR-T group with binder 16 started to develop GvHD related symptoms including scaly and NAI-1540294631v3 240 inflamed skin at extremities, while mice in CAR-T group with binder 8 didn’t exhibit any similar symptoms. All these data suggest that binder 8 and binder 16 CAR-T cells could efficiently control tumor growth in vivo. Lower doses can be further assessed to explore the antitumor potency of the CAR-T and mitigate GvHD issues. Example 7. In Vivo Safety and Effective Dose of GPC3-Targeted CAR T Cells [00828] To evaluate the safety and effective dose of the GPC3-targeted CAR with binder 8 and binder 16, 6- to 8-week-old NOD/SCID/IL-2Rg-null (NSG) female mice were injected with 5 × 106 mCherry-luciferase overexpressing Huh-7D12 cells subcutaneously on day 0 followed by i.v. injection of 4 × 106, 2 × 106, 1 × 106 and 5 × 105 CAR-positive T cells. Tumor volume (length and width) was measured 3 times per week with caliper and calculated as volume = 0.5 × length × width2. Body weight was measured 3 times a week to assess the acute and severe toxicities associated with the treatment. Blood samples were collected on day 7, day 14 post CAR-T injection and termination day of the study. The presence of human CAR-T cells was assessed by flow cytometry and IFN-γ secretion was quantified by ELISA. All animals were sacrificed according to IACUC protocol standards. An experimental layout was illustrated in Figure 10. Further, peripheral blood from mice treated with CAR-T cells was collected on Day 51 and concentration of IFN- γ in the plasma was quantified by ELISA. [00829] In CAR-T groups with binder 8, all mice treated with the higher doses (4 × 106 and 2 × 106) survived until termination of the study, while 2 of 5 mice treated with the lower doses (1 × 106 and 0.5 × 106) died from tumor progression. See Figure 11 and Figure 12. Similarly, CAR-T groups with binder 16, all mice survived except for mice treated with 0.5 × 106 CAR-T cells, as 3 of 5 mice died from tumor progression. See Figure 11 and Figure 12. Tumor growth was controlled by CAR-T cells in all mice that survived throughout the study. See Figure 13. These data show that CAR-T with both binder 8 and binder 16 can efficiently control tumor growth in vivo in a dose dependent manner. [00830] In all dose groups, neither binder showed any signs of toxicity or GvHD, indicating that 4 × 106 CAR+ is well-tolerated in mouse model. Flow cytometry analysis showed that CAR- T cells with binder 8 expanded for the first 2 weeks post injection and decreased in cell number at the end of study, which is consistent with the typical CAR-T in vivo kinetics (data not shown). However, in CAR-T groups with binder 16, human T cell number kept increasing during the study period and plasma IFN-γ level remained high at the end of this study. See Figure 14. NAI-1540294631v3 241 These data show that CAR-T with binder 16 retained an activation status and expanded during the study, which might lead to safety issues moving to human clinical studies. In contrast, CAR- T cells comprising binder 8 demonstrated potent anti-tumor efficacy and showed a minimal risk of toxicity in vivo. Example 8. Restimulation Assay with Huh7 Tumor Cells [00831] To further evaluate cytotoxicity activities of the GPC3-targeted CAR T cells described in Example 4, the GPC3-targeted CAR T cells were added to a Huh7 tumor monolayer at a 1:1 ratio every 72 hours. The cytotoxicity and proliferation was measured by bioluminescence and flow cytometry respectively. [00832] The results of the restimulation assay indicate that the GPC3-targeted CAR T cells with binders 1, 2, 4, 8, 9, and 16 (corresponding to 1_28z1XX, 2_28z1XX, 4_28z1XX, 8_28z1XX, 9_28z1XX, and 16_28z1XX) can repeatedly kill tumor cells and continue to proliferate in response to antigen, respectively. As shown in Figure 15B, the GPC3-targeted CAR T cells exhibit higher cytotoxicity over four restimulation cycles compared to GC33 and Y035 CAR T cells. As shown in Figure 15C, the GPC3-targeted CAR T cells with binders 1, 8, 9 and 16 show higher CAR T-cell numbers over four restimulation cycles and continue to proliferate in response to Huh7 tumor cells. In sum, all 6 GPC3-targeted CAR T cells showed equal or increased cytotoxicity and proliferation compared to the GPC3-targeted CAR T cells, while the comparator Y035 CAR T cells showed poor cytotoxicity and proliferation when co- cultured in the restimulation assays. Example 9. In Vivo Antitumor Efficacy and Toxicity of GPC3-Targeted CAR T Cells to Different Tumor Cells [00833] Xenograft models of hepatocellular carcinoma (HCC) cell lines Huh-7 and PLC/PRF/5 were engrafted subcutaneously and orthotopically. Tumor cells Huh-7 or PLC/PRF/5 at the dose of 5.0e5, 1.0e6, and 1.5e6 were implanted, respectively, in the right flank subcutaneously. Alternatively, tumor cells Huh-7 or PLC/PRF/5 at the dose of 5.0e4, 1.0e5, and 2.0e5 were implanted orthotopically in the liver, respectively. The tumor growth was measured by BLI. Natural tumor progression was monitored without any CAR T-cell treatment (data not shown). The results indicate that Xenograft models for HCC cell lines Huh-7 and PLC/PRF/5 were established subcutaneously and orthotopically in the liver. NAI-1540294631v3 242 [00834] Specifically, an orthotopic liver xenograft model was established by injecting 5×104 GFP-luciferase expressing Huh-7 tumor cells into the liver of 6- to 8-week-old NSG female mice at day 0 followed by i.v injection of 2×105 or 5×105 GPC3-targeted CAR T cells. The hypothesis is that a liver orthotopic tumor xenograft will more closely represent a HCC disease model. The GPC3-targeted CAR T cells with binders 1, 9 and 16 (corresponding to 1_28z1XX, 9_28z1XX, and 16_28z1XX) were chosen to represent a range of affinities and epitope binding scFv binders. All CAR T-cells showed anti-tumor efficacy at both doses. See Figure 16. Four mice treated with 2×10516_28z1XX and 2 mice treated with GC33 CAR T cells had long term tumor free survival. Mice treated with the higher dose of 5×1059_28z1XX and 16_28z1XX CAR T cells had long term anti-tumor activity equal to or greater than GC33_28z1XX. [00835] To further evaluate the antitumor efficacy and toxicity of the GPC3-targeted CAR T cells with the scFv binders in vivo, a higher dose of 5 × 105 Huh7 tumor cells were implanted subcutaneously. In brief, 6- to 8-week-old NSG female mice (n=5) were injected with 5e5 GFP- luciferase expressing Huh-7 cells subcutaneously on day 0 followed by i.v. injection of 2×105 CAR T-cells on day 7. Tumor burden was measured every 7 days using bioluminescent imaging (BLI). All novel GPC3 binders showed greater anti-tumor activity compared to GC33 and untransduced T cells. See Figure 17. Two mice treated with 8_28z1XX CAR T cells achieved sustained tumor free survival until termination of the study. This data suggest that binder 8- CAR-T cells can efficiently control tumor growth in vivo. Higher doses can be further assessed to explore the antitumor potency of the CAR-T and achieve tumor free survival. [00836] To evaluate the antitumor efficacy and toxicity of the GPC3-targeted CAR T cells in vivo, 6- to 8-week-old NOD/SCID/IL-2Rg-null (NSG) female mice (n=5) were injected with 5×105 GFP-luciferase expressing Nalm6-GPC3+ cells i.v on day 0 followed by i.v. injection of 2×105 CAR T-cells on day 4. Tumor burden was measured every 7 days using bioluminescent imaging (BLI). All novel GPC3 binders showed greater anti-tumor activity compared to GC33 and untransduced (UTD) T cells and improved survival of mice over time. See Figure 18. Mice treated with 16_28z1XX CAR T-cells effectively controlled tumor growth in 3 out of 5 mice comparable to mice treated with CD19 CAR T-cells 1928z1XX. These data suggest that binder 16-CAR-T cells could efficiently control tumor growth in vivo. Higher doses can be further assessed to explore the antitumor potency of the CAR-T and achieve tumor free survival. NAI-1540294631v3 243 Example 10. Bioinformatic Analysis on GPC3 Expression in HCC [00837] Proteomic (bulk) expression of GPC3 was analyzed using the data pulled from the Chinese Human Proteome Project (CNHPP) (Jiang et al., 2019, Nature 567: 257–261) and the Clinical Proteomic Tumor Analysis Consortium (CPTAC) (Gao et al., 2019, Cell 179 (2): 561 - 577.e22). The CNHPP characterized 110 paired tumor and non-tumor tissues of early-stage hepatocellular carcinoma related to hepatitis B virus infection using proteomic and phospho- proteomic profiling. The CPTAC performed the first proteogenomic characterization of HBV- related hepatocellular carcinoma using paired tumor and adjacent liver tissues from 159 patients, revealing insights into multi-omics consistency, key signaling pathways, and liver-specific metabolic reprogramming. In the analysis on proteomic (bulk) expression of GPC3, log2(fold change) of protein data was calculated by comparing tumor and normal liver samples. Molecules were filtered to show only those found on the cell surface (The UniProt Consortium et al., 2023, Nucleic Acids Research 51 (D1): D523–D531; Thul et al., 2017, Science 356, eaal3321; Gene Ontology Consortium et al., 2023, Genetics 224, 1; Binder et. al., 2014, Database 2014) and over-expressed in tumor compared to liver. Log2FC of the filtered molecules were percentile ranked and their distribution plotted. Density plots were created to visualize the distribution of protein expression levels. [00838] Transcriptomic (bulk) expression of GPC3 was analyzed using the data pulled from The Cancer Genome Atlas (TCGA) and the CPTAC. The TCGA aimed to comprehensively analyze 363 HCC cases using whole-exome sequencing, DNA copy number analyses, and 196 HCC cases using DNA methylation, RNA, miRNA, and proteomic expression, identifying significantly mutated genes and molecular subtypes (Ally et al., 2017, Cell 169 (7): 1327 - 1341.e23). The goal for using the HCC CPTAC data was the same as mentioned in the analysis of the proteomic (bulk) expression of GPC3. In the analysis on transcriptomic (bulk) expression of GPC3, log2(fold change) of transcript data was calculated by comparing tumor and normal liver samples. Transcripts were filtered to show only those coding for membrane-bound proteins and overexpressed in tumor compared to liver. Log2FC of the filtered transcripts were percentile ranked and their distribution plotted. Density plots were created to visualize the distribution of transcript expression levels. [00839] As shown in Figure 19A, GPC3 is identified as the most over-expressed membrane- bound protein when comparing tumor versus normal liver samples in both the CNHPP and NAI-1540294631v3 244 CPTAC datasets. As shown in Figure 19B, GPC3 is among the most highly over-expressed transcripts when comparing tumor versus normal liver samples in both the TCGA and CPTAC datasets. [00840] Although the presently disclosed subject matter and certain of its advantages have been described in detail, it should be understood that various changes, substitutions and alterations can be made herein without departing from the spirit and scope of the disclosure. Moreover, the scope of the present application is not intended to be limited to the particular embodiments of the process, machine, manufacture, and composition of matter, and methods described in the specification. As one of ordinary skill in the art will readily appreciate from the disclosure of the presently disclosed subject matter, processes, machines, manufacture, compositions of matter, or methods, presently existing or later to be developed that perform substantially the same function or achieve substantially the same result as the corresponding embodiments described herein may be utilized according to the presently disclosed subject matter. Accordingly, the appended claims are intended to include within their scope such processes, machines, manufacture, compositions of matter, or methods. [00841] Various patents, patent applications, publications, product descriptions, protocols, and sequence accession numbers are cited throughout this application, the disclosure of which are incorporated herein by reference in their entireties for all purposes. NAI-1540294631v3 245

Claims

WHAT IS CLAIMED IS: 1. A chimeric antigen receptor (CAR), comprising an extracellular domain that binds to GPC3, a transmembrane domain, and an intracellular domain, wherein the extracellular domain comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein a) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 30; b) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 47, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 48; c) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 62, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 63; d) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 75, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 76; e) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 89, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 90; f) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 101, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 102; g) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 119, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 120; h) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 137, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 138; 1540294631v3 246 i) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 155, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 156; j) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 170, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 171; k) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 183, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 184; l) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 201, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 202; m) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 215, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 216; n) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 233, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 234; o) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 246, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 247; or p) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 262, and the VL comprises a VL CDR1, a VL NAI-1540294631v3 247 CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 263.
2. The CAR of claim 1, wherein the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system, the Chothia numbering system, the AbM numbering system, the Contact numbering system, or the IMGT® Information System.
3. The CAR of claim 1, wherein the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system.
4. The CAR of claim 1, wherein: a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof; b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; c) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof; and the NAI-1540294631v3 248 VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; d) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; e) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; f) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; g) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the NAI-1540294631v3 249 amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof; h) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof; i) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof; j) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof; k) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence NAI-1540294631v3 250 set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; l) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof; m) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof; n) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof; NAI-1540294631v3 251 o) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof; or p) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof.
5. The CAR of any one of claims 1-4, wherein: a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28; b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; c) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a NAI-1540294631v3 252 CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; d) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; e) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; f) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; g) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118; h) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a NAI-1540294631v3 253 CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136; i) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154; j) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169; k) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; l) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200; m) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising NAI-1540294631v3 254 the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169; n) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232; o) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118; or p) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
6. The CAR of claim 5, wherein the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
7. The CAR of claim 5, wherein the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: NAI-1540294631v3 255 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
8. The CAR of any one of claims 1-7, wherein VH comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262.
9. The CAR of any one of claims 1-8, wherein VH comprises the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262.
10. The CAR of any one of claims 1-9, wherein VL comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263.
11. The CAR of any one of claims 1-10, wherein VL comprises the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263. NAI-1540294631v3 256
12. The CAR of any one of claims 1-11, wherein: a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; b) the VH comprises the amino acid sequence set forth in SEQ ID NO: 47; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 48; c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 62; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 63; d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; e) the VH comprises the amino acid sequence set forth in SEQ ID NO: 89; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 90; f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 101; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 102; g) the VH comprises the amino acid sequence set forth in SEQ ID NO: 119; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 120; h) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; i) the VH comprises the amino acid sequence set forth in SEQ ID NO: 155; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 156; j) the VH comprises the amino acid sequence set forth in SEQ ID NO: 170; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 171; k) the VH comprises the amino acid sequence set forth in SEQ ID NO: 183; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 184; l) the VH comprises the amino acid sequence set forth in SEQ ID NO: 201; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 202; m) the VH comprises the amino acid sequence set forth in SEQ ID NO: 215; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 216; n) the VH comprises the amino acid sequence set forth in SEQ ID NO: 233; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 234; o) the VH comprises the amino acid sequence set forth in SEQ ID NO: 246; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 247; or NAI-1540294631v3 257 p) the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263.
13. The CAR of claim 12, wherein: a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; b) the VH comprises the amino acid sequence set forth in SEQ ID NO: 47; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 48; c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; e) the VH comprises the amino acid sequence set forth in SEQ ID NO: 155; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 156; or f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263.
14. The CAR of claim 13, wherein the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138.
15. The CAR of claim 13, wherein the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263.
16. The CAR of any one of claims 1-15, wherein the extracellular domain comprises a variable fragment (Fv), a single-chain variable fragment (scFv), a Fab, or a F(ab)2.
17. The CAR of claim 16, wherein the extracellular domain comprises an scFv.
18. The CAR of any one of claims 1-17, wherein the extracellular domain comprises a linker between the VH and the VL.
19. The CAR of claim 18, wherein the linker consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4. NAI-1540294631v3 258
20. The CAR of any one of claims 1-19, wherein the VH and the VL are positioned from the N- to the C-terminus: VL-VH.
21. The CAR of any one of claims 1-19, wherein the VH and the VL are positioned from the N- to the C-terminus: VH-VL.
22. The CAR of any one of claims 1-21, wherein the extracellular domain comprises or is an scFv, which comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO: 264, or SEQ ID NO: 265.
23. The CAR of claim 22, wherein the extracellular domain comprises or is an scFv, which comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 264, or SEQ ID NO: 265.
24. The CAR of any one of claims 1-23, wherein the transmembrane domain comprises a CD8 polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, or a BTLA polypeptide.
25. The CAR of claim 24, wherein the transmembrane domain comprises a CD28 polypeptide.
26. The CAR of claim 25, wherein the CD28 polypeptide comprises the amino acid sequence set forth in amino acids 154 to 179 of SEQ ID NO: 308.
27. The CAR of any one of claims 1-26, wherein the intracellular domain comprises a CD3ζ polypeptide.
28. The CAR of claim 27, wherein the CD3ζ polypeptide is a modified CD3ζ polypeptide. NAI-1540294631v3 259
29. The CAR of claim 28, wherein the modified CD3ζ polypeptide comprises a native ITAM1, an ITAM2 variant consisting of two loss-of-function mutations, and an ITAM3 consisting of two loss-of-function mutations.
30. The CAR of claim 29, wherein the native ITAM1 consists of the amino acid sequence set forth in SEQ ID NO: 313.
31. The CAR of claim 29, wherein the ITAM2 variant consists of the amino acid sequence set forth in SEQ ID NO: 319.
32. The CAR of claim 29, wherein the ITAM3 variant consists of the amino acid sequence set forth in SEQ ID NO: 323.
33. The CAR of any one of claims 28-32, wherein the modified CD3ζ polypeptide comprises or consists of the amino acid sequence set forth in SEQ ID NO: 325.
34. The CAR of any one of claims 1-33, wherein the intracellular domain further comprises at least one co-stimulatory signaling region.
35. The CAR of claim 34, wherein the at least one co-stimulatory signaling region comprises at least one intracellular signaling domain of a co-stimulatory molecule.
36. The CAR of claim 35, wherein the co-stimulatory molecule is selected from the group consisting of CD28, 4-1BB, OX40, ICOS, DAP-10, CD27, CD28, CD30, CD40, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, CD8, CD4, B2C, CD80, CD86, DAP10, DAP12, MyD88, BTNL3, and NKG2D, and combinations thereof.
37. The CAR of claim 36, wherein the co-stimulatory molecule is CD28.
38. The CAR of any one of claims 34-37, wherein the at least one co-stimulatory signaling region comprises amino acids 180 to 220 of SEQ ID NO: 308.
39. The CAR of any one of claims 1-38, further comprising a signal peptide. NAI-1540294631v3 260
40. The CAR of claim 39, wherein the signal peptide comprises the amino acid sequence set forth in SEQ ID NO: 278.
41. The CAR of any one of claims 1-40, further comprising a self-cleaving P2A peptide.
42. The CAR of claim 41, wherein the self-cleaving P2A peptide comprises the amino acid sequence set forth in SEQ ID NO: 329.
43. The CAR of any one of claims 1-42, comprising the amino acid sequence set forth in SEQ ID NO: 333, SEQ ID NO: 335, SEQ ID NO: 337, SEQ ID NO: 339, SEQ ID NO: 341, SEQ ID NO: 343, SEQ ID NO: 345, SEQ ID NO: 347, SEQ ID NO: 349, SEQ ID NO: 351, SEQ ID NO: 353, or SEQ ID NO: 355.
44. The CAR of any one of claims 1-43, wherein the CAR is expressed from a vector.
45. The CAR of claim 44, wherein the vector is a viral vector.
46. The CAR of claim 45, wherein the viral vector is a retroviral vector.
47. A nucleic acid encoding the CAR of any one of claims 1-46.
48. The nucleic acid of claim 47, comprising the nucleotide sequence set forth in SEQ ID NO: 334, SEQ ID NO: 336, SEQ ID NO: 338, SEQ ID NO: 340, SEQ ID NO: 342, SEQ ID NO: 344, SEQ ID NO: 346, SEQ ID NO: 348, SEQ ID NO: 350, SEQ ID NO: 352, SEQ ID NO: 354, or SEQ ID NO: 356.
49. The nucleic acid of claim 47 or 48, further comprising a promoter that is operably linked to the CAR.
50. The nucleic acid of claim 49, wherein the promoter is endogenous or exogenous.
51. The nucleic acid of claim 50, wherein the exogenous promoter is selected from the group consisting of an elongation factor (EF)-1 promoter, a cytomegalovirus immediate-early promoter (CMV) promoter, a simian virus 40 early promoter (SV40) promoter, a phosphoglycerate kinase (PGK) promoter, a metallothionein promoter, and Ubiquitin C promoter. NAI-1540294631v3 261
52. The nucleic acid of claim 50, wherein the endogenous promoter is selected from a TCR alpha promoter, a TCR beta promoter, and a beta 2-microglobulin promoter.
53. The nucleic acid of claim 49, wherein the promoter is an inducible promoter.
54. The nucleic acid of claim 53, wherein the inducible promoter is selected from the group consisting of a NFAT transcriptional response element (TRE) promoter, a CD69 promoter, a CD25 promoter, an IL-2 promoter, a 4-1BB promoter, a PD1 promoter, and a LAG3 promoter.
55. A vector comprising the nucleic acid of any one of claims 47-54.
56. The vector of claim 55, wherein the vector is a viral vector.
57. The vector of claim 56, wherein the viral vector is a retroviral vector.
58. A cell comprising the CAR of any one of claims 1-46, the nucleic acid of any one of claims 47-54, or the vector of any one of claims 55-57.
59. The cell of claim 58, wherein the cell is transduced with the CAR, the nucleic acid, or the vector.
60. The cell of claim 58 or claim 59, wherein the CAR is constitutively expressed on the surface of the cell.
61. The cell of any one of claims 58-60, wherein the cell is an immunoresponsive cell or an immune effector cell.
62. The cell of any one of claims 58-61, wherein the cell is a cell of the lymphoid lineage or a cell of the myeloid lineage.
63. The cell of any one of claims 58-62, wherein the cell is selected from the group consisting of a T cell, a B cell, a Natural Killer (NK) cell, macrophage, innate lymphoid cell (ILC), a cytokine induced killer (CIK) cell, a lymphokine activated killer (LAK) cell, a stem cell from which a lymphoid cell may be differentiated, a stem cell from which a myeloid cell may be differentiated, or a combination thereof. NAI-1540294631v3 262
64. The cell of claim 63, wherein the cell is a T cell.
65. The cell of claim 63 or claim 64, wherein the T cell is selected from the group consisting of helper T cells, cytotoxic T cells, memory T cells, regulatory T cells, tumor-infiltrating lymphocytes (TILs), Natural Killer T cells, mucosal associated invariant T cells, αβ T cells, and γδ T cells.
66. The cell of claim 63, wherein the cell is a NK cell.
67. The cell of claim 66, wherein the NK cell is derived from a stem cell.
68. The cell of claim 63 or 67, wherein the stem cell is a pluripotent stem cell.
69. The cell of claim 68, wherein the pluripotent stem cell is an embryoid stem cell or an induced pluripotent stem cell.
70. An anti-GPC3 antibody or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein a) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 30; b) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 47, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 48; c) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 62, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 63; d) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 75, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 76; e) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 89, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 90; NAI-1540294631v3 263 f) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 101, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 102; g) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 119, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 120; h) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 137, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 138; i) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 155, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 156; j) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 170, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 171; k) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 183, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 184; l) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 201, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 202; m) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 215, and the VL comprises a VL CDR1, a VL NAI-1540294631v3 264 CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 216; n) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 233, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 234; o) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 246, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 247; or p) the VH comprises a VH CDR1, a VH CDR2, and a VH CDR3 of a VH comprising the amino acid sequence set forth in SEQ ID NO: 262, and the VL comprises a VL CDR1, a VL CDR2, and a VL CDR3 of a VL comprising the amino acid sequence set forth in SEQ ID NO: 263.
71. The antibody or antigen-binding fragment thereof of claim 70, wherein the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system, the Chothia numbering system, the AbM numbering system, the Contact numbering system, or the IMGT® Information System.
72. The antibody or antigen-binding fragment thereof of claim 71, wherein the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 are identified according to the Kabat numbering system.
73. The antibody or antigen-binding fragment thereof of any one of claims 70-72, wherein: a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ NAI-1540294631v3 265 ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28 or a conservative modification thereof; b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; c) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; d) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; e) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a NAI-1540294631v3 266 conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; f) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; g) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof; h) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136 or a conservative modification thereof; i) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151 or a conservative modification thereof; and NAI-1540294631v3 267 the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154 or a conservative modification thereof; j) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof; k) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46 or a conservative modification thereof; l) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200 or a conservative modification thereof; m) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167 or a conservative modification thereof, and a CDR3 comprising the NAI-1540294631v3 268 amino acid sequence set forth in SEQ ID NO: 168 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169 or a conservative modification thereof; n) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232 or a conservative modification thereof; o) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118 or a conservative modification thereof; or p) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260 or a conservative modification thereof; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230 or a conservative modification thereof, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231 or a conservative modification thereof, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261 or a conservative modification thereof.
74. The antibody or antigen-binding fragment thereof of any one of claims 70-73, wherein: NAI-1540294631v3 269 a) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 28; b) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; c) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 59, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 61; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; d) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 23, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 60, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 25; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 74, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; e) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 87, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 88, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; NAI-1540294631v3 270 f) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 42, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 43; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 44, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 45, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; g) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 114, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118; h) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136; i) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 149, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 150, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 151; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 152, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 153, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 154; j) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169; NAI-1540294631v3 271 k) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 41, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 24, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 182; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 26, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 27, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 46; l) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 195, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 196, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 197; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 198, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 199, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 200; m) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 213, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 167, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 168; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 214, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 169; n) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 227, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 228, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 229; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 232; o) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 113, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 245, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 115; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 116, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 117, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 118; or NAI-1540294631v3 272 p) the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
75. The antibody or antigen-binding fragment thereof of claim 74, wherein the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 131, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 132, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 133; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 134, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 135, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 136.
76. The antibody or antigen-binding fragment thereof of claim 74, wherein the VH comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 258, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 259, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 260; and the VL comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 230, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 231, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 261.
77. The antibody or antigen-binding fragment thereof of any one of claims 70-76, wherein VH comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262. NAI-1540294631v3 273
78. The antibody or antigen-binding fragment thereof of any one of claims 70-77, wherein VH comprises the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262.
79. The antibody or antigen-binding fragment thereof of any one of claims 70-78, wherein VL comprises an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% identical or homologous to the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263.
80. The antibody or antigen-binding fragment thereof of any one of claims 70-79, wherein VL comprises the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263.
81. The antibody or antigen-binding fragment thereof of any one of claims 70-80, wherein: a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; b) the VH comprises the amino acid sequence set forth in SEQ ID NO: 47; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 48; c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 62; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 63; d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; e) the VH comprises the amino acid sequence set forth in SEQ ID NO: 89; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 90; NAI-1540294631v3 274 f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 101; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 102; g) the VH comprises the amino acid sequence set forth in SEQ ID NO: 119; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 120; h) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; i) the VH comprises the amino acid sequence set forth in SEQ ID NO: 155; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 156; j) the VH comprises the amino acid sequence set forth in SEQ ID NO: 170; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 171; k) the VH comprises the amino acid sequence set forth in SEQ ID NO: 183; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 184; l) the VH comprises the amino acid sequence set forth in SEQ ID NO: 201; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 202; m) the VH comprises the amino acid sequence set forth in SEQ ID NO: 215; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 216; n) the VH comprises the amino acid sequence set forth in SEQ ID NO: 233; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 234; o) the VH comprises the amino acid sequence set forth in SEQ ID NO: 246; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 247; or p) the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263.
82. The antibody or antigen-binding fragment thereof of claim 81, wherein: a) the VH comprises the amino acid sequence set forth in SEQ ID NO: 29; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 30; b) the VH comprises the amino acid sequence set forth in SEQ ID NO: 47; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 48; c) the VH comprises the amino acid sequence set forth in SEQ ID NO: 75; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 76; d) the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138; NAI-1540294631v3 275 e) the VH comprises the amino acid sequence set forth in SEQ ID NO: 155; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 156; or f) the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263.
83. The antibody or antigen-binding fragment thereof of claim 82, wherein the VH comprises the amino acid sequence set forth in SEQ ID NO: 137; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 138.
84. The antibody or antigen-binding fragment thereof of claim 82, wherein the VH comprises the amino acid sequence set forth in SEQ ID NO: 262; and the VL comprises the amino acid sequence set forth in SEQ ID NO: 263,
85. The antibody or antigen-binding fragment thereof of any one of claims 70-84, wherein the antigen-binding fragment thereof is a variable fragment (Fv), a single-chain variable fragment (scFv), a Fab, or a F(ab)2.
86. The antibody or antigen-binding fragment thereof of claim 85, wherein the antigen- binding fragment thereof is an scFv.
87. The antibody or antigen-binding fragment thereof of claim 86, wherein the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 121, SEQ ID NO: 122, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 235, SEQ ID NO: 236, SEQ ID NO: 248, SEQ ID NO: 249, SEQ ID NO: 264, or SEQ ID NO: 265.
88. The antibody or antigen-binding fragment thereof of claim 87, wherein the scFv comprises or consists of the amino acid sequence set forth in SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 264, or SEQ ID NO: 265. NAI-1540294631v3 276
89. The antibody or antigen-binding fragment thereof of any one of claims 70-88, wherein the VH is connected to the VL via a linker.
90. The antibody or antigen-binding fragment thereof of claim 89, wherein the linker consists of the amino acid sequence set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.
91. The antibody or antigen-binding fragment thereof of any one of claims 70-90, wherein the VH and the VL are positioned from the N- to the C-terminus: VL-VH.
92. The antibody or antigen-binding fragment thereof of any one of claims 70-90, wherein the VH and the VL are positioned from the N- to the C-terminus: VH-VL.
93. The antibody or antigen-binding fragment thereof of any one of claims 70-92, wherein the antibody comprises a human variable region framework region.
94. The antibody or antigen-binding fragment thereof of any one of claims 70-83, wherein the antibody or antigen-binding fragment thereof comprises a Fc region.
95. The antibody or antigen-binding fragment thereof of claim 94, wherein the Fc region is a IgG1 Fc region.
96. The antibody or antigen-binding fragment thereof of claim 94 or 95, wherein the Fc region comprises the amino acid sequence set forth in SEQ ID NO: 357.
97. The antibody or antigen-binding fragment thereof of any one of claims 70-96, which is a fully human or an antigen-binding fragment thereof, a chimeric antibody or an antigen-binding fragment thereof, or a humanized antibody or an antigen-binding fragment thereof.
98. An antibody or an antigen-binding fragment thereof that competes with the antibody or antigen-binding fragment thereof of any one of claims 70-97 for binding to GPC3.
99. An antibody or an antigen-binding fragment thereof that binds to essentially the same epitope region on GPC3 as the antibody or antigen-binding fragment thereof of any one of claims 70-97. NAI-1540294631v3 277
100. A conjugate comprising the antibody or antigen-binding fragment thereof of any one of claims 70-97.
101. The conjugate of claim 100, wherein the antibody or antigen-binding fragment thereof is linked to a therapeutic agent, a detectable agent, or a diagnostic agent.
102. The conjugate of claim 101, wherein the therapeutic agent is a chemotherapeutic agent, a cytotoxin, or a drug.
103. The conjugate of any one of claims 100-102, which is an immunoconjugate.
104. A multispecific molecule comprising the antibody or the antigen-binding fragment thereof of any one of claims 70-97, linked to a second functional moiety.
105. The multispecific molecule of claim 104, wherein the second functional moiety has a different binding specificity than the antibody or antigen binding fragment thereof.
106. A nucleic acid that encodes the antibody or antigen-binding fragment thereof of any one of claims 70-97.
107. The nucleic acid of claim 106, which comprises a first polynucleotide that encodes a VH comprising the amino acid sequence set forth in SEQ ID NO: 29, SEQ ID NO: 47, SEQ ID NO: 62, SEQ ID NO: 75, SEQ ID NO: 89, SEQ ID NO: 101, SEQ ID NO: 119, SEQ ID NO: 137, SEQ ID NO: 155, SEQ ID NO: 170, SEQ ID NO: 183, SEQ ID NO: 201, SEQ ID NO: 215, SEQ ID NO: 233, SEQ ID NO: 246, or SEQ ID NO: 262; and/or a second polynucleotide that encodes a VL comprising the amino acid sequence set forth in SEQ ID NO: 30, SEQ ID NO: 48, SEQ ID NO: 63, SEQ ID NO: 76, SEQ ID NO: 90, SEQ ID NO: 102, SEQ ID NO: 120, SEQ ID NO: 138, SEQ ID NO: 156, SEQ ID NO: 171, SEQ ID NO: 184, SEQ ID NO: 202, SEQ ID NO: 216, SEQ ID NO: 234, SEQ ID NO: 247, or SEQ ID NO: 263.
108. The nucleic acid of claim 107, wherein the first polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO: 33, SEQ ID NO: 51, SEQ ID NO: 66, SEQ ID NO: 79, SEQ ID NO: 93, SEQ ID NO: 105, SEQ ID NO: 123, SEQ ID NO: 141, SEQ ID NO: 159, SEQ ID NO: 174, SEQ ID NO: 187, SEQ ID NO: 205, SEQ ID NO: 219, SEQ ID NO: 237, SEQ ID NO: 250, or SEQ ID NO: 266. NAI-1540294631v3 278
109. The nucleic acid of claim 107 or 108, wherein the second polynucleotide comprises the nucleotide sequence set forth in SEQ ID NO: 35, SEQ ID NO: 53, SEQ ID NO: 68, SEQ ID NO: 81, SEQ ID NO: 95, SEQ ID NO: 107, SEQ ID NO: 125, SEQ ID NO: 143, SEQ ID NO: 161, SEQ ID NO: 176, SEQ ID NO: 189, SEQ ID NO: 207, SEQ ID NO: 221, SEQ ID NO: 239, SEQ ID NO: 252, or SEQ ID NO: 268.
110. A vector comprising the nucleic acid of any one of claims 106-109.
111. The vector of claim 110, which is an expression vector.
112. A host cell comprising the vector of claim 110 or 111.
113. A method of producing an anti-GPC3 antibody or an antigen-binding fragment thereof, wherein the method comprises culturing the host cell of claim 111 under conditions to induce expression of the antibody or antigen-binding fragment thereof from the host cell.
114. A method for detecting GPC3 in a whole cell or tissue, comprising: a) contacting a cell or tissue with the antibody or antigen-binding fragment thereof of any one of claims 70-97, wherein the antibody or antigen-binding fragment thereof comprises a detectable label; and b) determining the amount of the labeled antibody or antigen-binding fragment thereof bound to said cell or tissue by measuring the amount of detectable label associated with the cell or tissue, wherein the amount of bound antibody or antigen-binding fragment thereof indicates the amount of GPC3 in the cell or tissue.
115. A composition comprising the cell of any one of claims 58-69, the antibody or antigen- binding fragment thereof of any one of claims 70-97, the conjugate of any one of claims 100- 103, or the multispecific molecule of claim 104 or claim 105.
116. The composition of claim 115, which is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.
117. The composition of claim 115 or claim 116, comprising between about 1 × 105 and about 5 × 108 cells. NAI-1540294631v3 279
118. A method of treating a disease or disorder associated with GPC3 in a subject, comprising administering to the subject the cell of any one of claims 58-69, the antibody or antigen-binding fragment thereof of any one of claims 70-97, the conjugate of any one of claims 100-103, the multispecific molecule of claim 104 or claim 105, or the composition of any one of claims 115- 117.
119. The method of claim 118, wherein the disease or disorder is a tumor.
120. The method of claim 118 or claim 119, wherein the disease or disorder is cancer.
121. The method of any one of claims 118-120, wherein the disease or disorder is selected from the group consisting of hepatocellular carcinoma, hepatoblastoma, lung squamous cell carcinoma, ovarian yolk sac tumor, melanoma, and urothelial carcinoma.
122. The method of claim 121, wherein the disease or disorder is hepatocellular carcinoma.
123. The method of any one of claims 118-122, wherein the subject is a human subject.
124. A method for producing a cell comprising the CAR of any one of claims 1-46, comprising introducing into a cell a nucleic acid that encodes the CAR of any one of claims 1- 46.
125. The cell of any one of claims 58-69, the antibody or antigen-binding fragment thereof of any one of claims 70-97, the conjugate of any one of claims 100-103, the multispecific molecule of claim 104 or claim 105, or the composition of any one of claims 115-117 for use in a therapy.
126. The cell of any one of claims 58-69, the antibody or antigen-binding fragment thereof of any one of claims 70-97, the conjugate of any one of claims 100-103, the multispecific molecule of claim 104 or claim 105, or the composition of any one of claims 115-117 for use in a method for treating a disease or disorder associated with GPC3 in a subject.
127. The use of claim 126, wherein the disease or disorder is a tumor.
128. The use of claim 126 or claim 127, wherein the disease or disorder is cancer. NAI-1540294631v3 280
129. The use of any one of claims 126-128, wherein the disease or disorder is selected from the group consisting of hepatocellular carcinoma, hepatoblastoma, lung squamous cell carcinoma, ovarian yolk sac tumor, melanoma, and urothelial carcinoma.
130. The use of claim 129, wherein the disease or disorder is hepatocellular carcinoma.
131. The use of any one of claims 126-130, wherein the subject is a human subject. NAI-1540294631v3 281
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