[go: up one dir, main page]

WO2025085512A1 - Polythérapies avec des anticorps ciblant ulbp2 pour traiter des cancers - Google Patents

Polythérapies avec des anticorps ciblant ulbp2 pour traiter des cancers Download PDF

Info

Publication number
WO2025085512A1
WO2025085512A1 PCT/US2024/051568 US2024051568W WO2025085512A1 WO 2025085512 A1 WO2025085512 A1 WO 2025085512A1 US 2024051568 W US2024051568 W US 2024051568W WO 2025085512 A1 WO2025085512 A1 WO 2025085512A1
Authority
WO
WIPO (PCT)
Prior art keywords
amino acid
seq
acid sequence
cancer
numbering
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/US2024/051568
Other languages
English (en)
Other versions
WO2025085512A9 (fr
Inventor
Stella Aviaty MARTOMO
Jeremy S. MYERS
Oksana SERGEEVA
Eric M. TAM
Jay Fine
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Evolveimmune Therapeutics Inc
Original Assignee
Evolveimmune Therapeutics Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Evolveimmune Therapeutics Inc filed Critical Evolveimmune Therapeutics Inc
Publication of WO2025085512A1 publication Critical patent/WO2025085512A1/fr
Publication of WO2025085512A9 publication Critical patent/WO2025085512A9/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2833—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against MHC-molecules, e.g. HLA-molecules
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • 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
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A61K2039/507—Comprising a combination of two or more separate antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7068—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines having oxo groups directly attached to the pyrimidine ring, e.g. cytidine, cytidylic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/243—Platinum; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/70539—MHC-molecules, e.g. HLA-molecules
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • This disclosure relates to pharmaceutical compositions of antibodies that bind ULBP2 and their use.
  • This disclosure relates to combination therapies for diseases associated with cells expressing ULBP2, such as bladder cancers.
  • This disclosure also relates to a method and kits for detecting a predisposition to, determining risk of, and guiding therapy for cancers.
  • UL16 binding protein 2 (ULBP2) is a cell surface glycoprotein related to MHC class I. ULBP2 functions as a stress-induced ligand for NKG2D receptor. ULBP2 encoded protein undergoes further processing to generate the mature protein that is either anchored to membrane via a glycosylphosphatidylinositol moiety or secreted.
  • NKG2D ligands are shown to be upregulated by a range of primary tumors, including lung, kidney, prostate, breast and colon cancer. Immune response induced by ULBP2- NKG2D may play an important role in the eradication of tumors by T and/or NK cells.
  • the present disclosure is based on the discovery that certain combinations of an anti-ULBP2 antibody or antigen binding fragment thereof (e.g., a bispecific antibody that binds ULBP2 and CD3) and a chemotherapy, radiotherapy, or an immune checkpoint inhibitor (e.g., pembrolizumab) increases efficacy relative to the anti- ULBP2 therapy alone in a subject having cancer (e.g., cancers with increased ULBP2 expression level).
  • an anti-ULBP2 antibody or antigen binding fragment thereof e.g., a bispecific antibody that binds ULBP2 and CD3
  • an immune checkpoint inhibitor e.g., pembrolizumab
  • the present disclosure provides a method of decreasing a population of cancer cells in a subject, the method comprising administering to the subject: a) an anti- ULBP2 antibody or antigen binding fragment thereof, and b) a chemotherapy agent, a radiation therapy, and/or an immune checkpoint inhibitor, wherein the plurality of cancer cells express UL16 binding protein 2 (ULBP2) and wherein the subject has cancer (e.g., a squamous cancer).
  • ULBP2 UL16 binding protein 2
  • the method results in a decrease of about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% in the number of ULBP2 expressing cancer cells in the subject relative to the number of ULBP2 expressing cancer cells in the subject prior to the administration of the anti-ULBP2 antibody or antigen binding fragment thereof and the chemotherapy agent and/or the radiation therapy.
  • the disclosure also provides a method for treating a human subject having a cancer (e.g., a basal squamous cancer) comprising the steps of: a) obtaining a biological sample from a human subject having cancer (e.g., a basal squamous cancer); b) detecting the expression levels of ULBP2; c) comparing the detected expression level of ULBP2 with a control level of ULBP2 expression; d) identifying the subject as a responder when the detected expression level of ULBP2 is greater than the control level of ULBP2 expression; and e) administering an anti-ULBP2 antibody or fragment thereof in an amount sufficient to alleviate a symptom of the cancer (e.g., basal squamous cancer) when the subject is identified as a responder.
  • a cancer e.g., a basal squamous cancer
  • the method comprises: i) detecting the expression levels of TROP2, PDL1, HLA-E, NECTIN4 and/or HER2; ii) comparing the detected expression levels of TROP2, PDL1, HLA-E, NECTIN4 and/or HER2 to a control levels of expression; iii) identifying the subject as a responder when the detected expression level of TROP2 is greater than the control level of TROP2 expression; the detected expression level of PDL1 is greater than the control level of PDL1 expression; the detected expression level of HLA-E is greater than the control level of HLA-E expression; the detected expression level of NECTIN4 is less than the control level of NECTIN4 expression; and/or the detected expression level of HER2 is less than the control level of HER2 expression.
  • the anti-ULBP2 antibody or fragment thereof comprises: a) a complementarity determining region 1 (VH2CDRI) comprising the amino acid sequence of SEQ ID NO: 428; a complementarity determining region 2 (VH2CDR2) comprising the amino acid sequence of SEQ ID NO: 430; a complementarity determining region 3 (VH2CDR3) comprising the amino acid sequence of SEQ ID NO: 432; a complementarity determining region 1 (VL2CDRI) comprising the amino acid sequence of SEQ ID NO: 433; a complementarity determining region 2 (VL2CDR2) comprising the amino acid sequence of SEQ ID NO: 434; and a complementarity determining region 3 (VL2CDR3) comprising the amino acid sequence of SEQ ID NO: 435; or b) a complementarity determining region 1 (VH2CDRI) comprising the amino acid sequence of SEQ ID NO: 5; a complementarity determining region 2 (VH2CDR2) comprising
  • the anti-ULBP2 antibody or fragment thereof is a bispecific antibody that specifically binds ULBP2 and CD3 or wherein the anti-ULBP2 antibody or fragment thereof is a bispecific antibody that specifically binds ULBP2 and PD1.
  • a CD58 polypeptide or a fragment thereof is fused to the C-terminus of a heavy chain polypeptide of the bispecific antibody.
  • the CD58 comprises the amino acid sequence of SEQ ID NO: 49.
  • the bispecific antibody comprises an anti-ULBP2 antibody or antigen binding fragment thereof comprising a light chain variable region (VL2) that comprises the amino acid sequence of SEQ ID NO: 1, and a heavy chain variable region (VH2) that comprises the amino acid sequence of SEQ ID NO: 629.
  • the anti-ULBP2 antibody or antigen binding fragment thereof comprises a light chain (L2) that comprises the amino acid sequence of SEQ ID NO: 67, or a heavy chain (H2) that comprises the amino acid sequence of SEQ ID NO: 621.
  • the bispecific antibody comprises an anti-CD3 antibody or antigen binding fragment thereof comprising a light chain variable region (VL1) that comprises the amino acid sequence of SEQ ID NO: 22, and a heavy chain variable region (VH1) that comprises the amino acid sequence of SEQ ID NO: 17.
  • VL1 light chain variable region
  • VH1 heavy chain variable region
  • the CD58 polypeptide or a fragment thereof is fused to the C-terminus of a heavy chain of the anti-CD3 antibody or antigen fragment thereof of the bispecific antibody.
  • the anti-CD3 antibody or antigen binding fragment thereof comprises a light chain (LI) comprising the amino acid sequence of SEQ ID NO: 69, and a heavy chain fused to the CD58 polypeptide (Hl) comprising the amino acid sequence of SEQ ID NO: 622.
  • LI light chain
  • Hl CD58 polypeptide
  • chemotherapy agents suitable for use in the present disclosure include but are not limited to a topoisomerase inhibitor, an anthracycline doxorubicin, a DNA break inducing antibiotic, a pyrimidine antagonist, or a platinum alkylating agent.
  • the radiation therapy is selected from the group consisting of 3D conformal radiation therapy (3DCRT), image-guided radiation therapy (IGRT), intensity modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), brachytherapy, intraoperative radiation therapy (IORT), stereotactic radiosurgery (SRS), proton therapy, MRI linear accelerator, and/or stereotactic body radiation therapy (SBRT).
  • 3DCRT 3D conformal radiation therapy
  • IGRT image-guided radiation therapy
  • IMRT intensity modulated radiation therapy
  • VMAT volumetric modulated arc therapy
  • IORT intraoperative radiation therapy
  • SRS stereotactic radiosurgery
  • proton therapy MRI linear accelerator
  • SBRT stereotactic body radiation therapy
  • the disclosure also provides a method for identifying a human subject having a cancer (e.g., basal squamous cancer) that is suitable for therapy with an anti-UL16 binding protein 2 (ULBP2) antibody or antigen binding fragment thereof comprising the steps of: a) obtaining a biological sample a human subject having cancer (e.g., basal squamous cancer); b) detecting the expression levels of ULBP2; c) comparing the detected expression level of ULBP2 with a control level of ULBP2 expression; and d) identifying the subject as a responder when the detected expression level of ULBP2 is greater than the control level of ULBP2 expression.
  • a cancer e.g., basal squamous cancer
  • ULBP2 anti-UL16 binding protein 2
  • the method further comprises: e) administering an anti-ULBP2 antibody or fragment thereof in an amount sufficient to alleviate a symptom of the cancer (e.g. basal squamous cancer) when the subject is identified as a responder.
  • a symptom of the cancer e.g. basal squamous cancer
  • the subject has a squamous cancer.
  • the squamous cancer is a basal squamous cancer.
  • the subject has a bladder cancer, a lung cancer, a head and neck cancer, a cervical cancer, esophageal cancer, anal cancer, vulvar cancer, penile cancer, thymic cancer, skin cancer cell, or adrenocortical cancer.
  • the subject has an adenocarcinoma.
  • the adenocarcinoma is lung adenocarcinoma, pancreatic adenocarcinoma, bladder adenocarcinoma, esophageal adenocarcinoma, or colon adenocarcinoma.
  • the subject has an EGFR activating mutation or an EGFR copy number alteration.
  • the subject has been previously treated with a tyrosine kinase inhibitors (TKI).
  • TKI tyrosine kinase inhibitors
  • the TKI is Osimertinib.
  • the TKI does not decrease ULBP2 expression level in the cancer.
  • the subject previously received TKI has relapsed cancer after TKI treatment.
  • the cancer is a metastatic cancer.
  • the subject with metastatic cancer has non-primary lesions in the liver, lung, bone, brain, or lymph node originated from a primary cancer.
  • the primary cancer is a squamous cancer or an adenocarcinoma.
  • the primary cancer is a squamous cancer comprising a bladder cancer, a lung cancer, a head and neck cancer, a cervical cancer, esophageal cancer, anal cancer, vulvar cancer, penile cancer, thymic cancer, skin cancer cell, or adrenocortical cancer.
  • the primary cancer is an adenocarcinoma including lung adenocarcinoma, pancreatic adenocarcinoma, bladder adenocarcinoma, esophageal adenocarcinoma, or colon adenocarcinoma.
  • the subject is previously administered immune checkpoint inhibitor.
  • the immune checkpoint inhibitor is an PD1/PDL1 inhibitor or a CTLA4 inhibitor.
  • the immune checkpoint inhibitor is pembrolizumab, nivolumab, atezolizumab, ipilimumab, or tremelimumab.
  • the immune checkpoint inhibitor does not decrease ULBP2 expression level in the cancer.
  • the subject is simultaneously administered or previously administered with a therapeutically effective amount of a therapeutic agent.
  • the therapeutic agent is a chemotherapy agent. Any suitable chemotherapy can be used in combination with an anti-ULBP2 antibody or antigen binding fragment described herein.
  • chemotherapy agents suitable for use in the present disclosure include but are not limited to a topoisomerase inhibitor, an anthracycline doxorubicin, a DNA break inducing antibiotic, a pyrimidine antagonist, or a platinum alkylating agent.
  • the therapeutic agent is a radiation therapy selected from the group consisting of 3D conformal radiation therapy (3DCRT), image-guided radiation therapy (IGRT), intensity modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), brachytherapy, intraoperative radiation therapy (IORT), stereotactic radiosurgery (SRS), proton therapy, MRI linear accelerator, and/or stereotactic body radiation therapy (SBRT).
  • 3DCRT 3D conformal radiation therapy
  • IGRT image-guided radiation therapy
  • IMRT intensity modulated radiation therapy
  • VMAT volumetric modulated arc therapy
  • IORT intraoperative radiation therapy
  • SRS stereotactic radiosurgery
  • proton therapy MRI linear accelerator
  • SBRT stereotactic body radiation therapy
  • the therapeutic agent is a CAR-T cell therapy, an immune checkpoint inhibitor, a co-stimulatory ligand or a cytokine.
  • the immune checkpoint inhibitor is pembrolizumab, nivolumab, atezolizumab, ipilimumab, or tremelimumab.
  • the anti-ULBP2 antibody or antigen binding fragment thereof and the chemotherapy agent, radiation therapy , and/or immune checkpoint inhibitor are administered sequentially. In some embodiments, the anti-ULBP2 antibody or antigen binding fragment thereof and the chemotherapy agent, radiation therapy, and/or immune checkpoint inhibitor are administered concurrently.
  • FIGS. 1A-1B are a series of heat maps showing the average levels of a set of luminal/adeno genes (top) and basal/squamous genes (bottom) in bladder and lung TCGA datasets and the Cancer Cell Line Encyclopedia (CCLE) when clustered using these same genes and others (immune infiltration, neuronal).
  • the levels in a subset of cell lines are shown and labeled by their category for bladder (luminal, luminal-infiltrated, or basal-squamous) or lung (mixed/infiltrated or basal-squamous).
  • FIG. 1A shows the bladder dataset
  • FIG. IB shows the lung dataset.
  • FIG. 2 is a heatmap depicting quantified immunoblots for luminal (FOXA1, PPARG, GATA3) and basal/squamous (KRT5, KRT6, and SERPINB3) markers at the protein level in several cell lines (shown on the x-axis). ULBP2 protein levels from flow cytometry antigen binding density are also shown for the same cell lines.
  • FIGS. 3A-3D are a series of graphs depicting gene levels in the bladder cancer TCGA dataset of the same clustered groups as defined and shown in FIG. 1.
  • FIG. 3A shows the target ULBP2.
  • FIG. 3B shows TROP2.
  • FIG. 3C shows HER2.
  • FIG. 3D shows NECTIN4.
  • p > 0.05 is denoted with *;
  • p ⁇ 0.01 is denoted with **;
  • p ⁇ 0.001 is denoted with ***;
  • p ⁇ 0.0001 is denoted with ****.
  • FIGS. 4A-4D are a series of graphs depicting gene levels in the bladder cancer TCGA dataset of the same clustered groups as defined and shown in FIG. 1.
  • FIG. 4A shows the luminal gene FOXA1.
  • FIG. 4B shows the luminal gene PPARG.
  • FIG. 4C shows the basal/squamous gene KRT5.
  • FIG. 4D shows SERPINB3.
  • FIGS. 5A-5D are a series of graphs depicting gene levels in the bladder cancer TCGA dataset of the same clustered groups as defined and shown in FIG. 1.
  • FIG. 5A shows the immune gene PDL1.
  • FIG. 5B shows the T cell gene PD1.
  • FIG. 5C shows the T cell gene CD8A.
  • FIG. 5D shows the T cell gene Granzyme B.
  • FIGS. 6A-6D are a series of graphs depicting gene levels in the lung cancer TCGA dataset bifurcated with either adenocarcinoma or squamous indication.
  • FIG. 6A shows the target ULBP2.
  • FIG. 6B shows TROP2.
  • FIG. 6C shows HER2.
  • FIG. 6D shows NECTIN4.
  • FIGS. 7A-7D are a series of graphs depicting gene levels in the lung cancer TCGA dataset bifurcated with either adenocarcinoma or squamous indication.
  • FIG. 7A shows the luminal gene FOXA1.
  • FIG. 7B shows the luminal gene PPARG.
  • FIG. 7C shows the basal/squamous gene KRT5.
  • FIG. 7D shows SERPINB3.
  • FIGS. 8A-8D are a series of graphs depicting gene levels in the lung cancer TCGA dataset bifurcated with either adenocarcinoma or squamous indication.
  • FIG. 8A shows the immune gene PDL1.
  • FIG. 8B shows the T cell gene PD1.
  • FIG. 8C shows the T cell gene CD8A.
  • FIG. 8D shows the T cell gene Granzyme B.
  • FIGS. 9A-9F are a series of graphs depicting T cell dependent cell cytotoxicity and cytokine release data in the bladder squamous cell line 5637 with anti-ULBP2 antibody EIP0820 and the negative control EIP0607 in combination with either pembrolizumab or the IgG4 isotype control.
  • FIG. 9A shows tumor killing at Day 5 with Donor 1 A.
  • FIG. 9B shows tumor killing at Day 5 with Donor 2A.
  • FIG. 9C shows tumor killing at Day 5 with Donor 3A.
  • FIG. 9D shows IFNg cytokine release at Day 2 with Donor 1A.
  • FIG. 9B shows IFNg cytokine release at Day 2 with Donor 2A.
  • FIG. 9A shows tumor killing at Day 5 with Donor 1 A.
  • FIGS. 10A-10B are a series of graphs depicting upregulation of ULBP2 and other markers (HER2, CD58, PDL1, TROP2, and B7H4) with a series of chemotherapy agents (bleomycin, cisplatin, docetaxel, doxorubicin, gemcitabine, irinotecan, and vinblastine) at their EC50 concentrations.
  • FIG. 10A shows the bladder squamous cell line 5637.
  • FIG. 10B shows the lung adenosquamous cell line A549.
  • FIG. 11 is a graph depicting T cell dependent cell cytotoxicity data in 5637 cells with EIP0820 and the negative control EIP0607 in combination with irinotecan chemotherapy.
  • FIG. 12 is a graph depicting T cell dependent cell cytotoxicity in squamous bladder cancer 5637 cell with EIP0820 and the negative control EIP0614 in combination with gemcitabine/cisplatin (“Cis/Gem”).
  • FIG. 13 is a graph depicting gene levels in an esophageal cancer dataset from The Cancer Genome Atlas (TCGA), bifurcated by either adenocarcinoma or squamous indication.
  • FIGS. 14A -C depict associations between ULBP2 expression, basal-squamous molecular subtype, and CD8 T cell infiltration, as outlined in Table 14.
  • FIG. 14A shows ULBP2 expression versus basal/squamous subtype.
  • FIG. 14B shows ULBP2 expression versus CD8 T cell infiltration.
  • FIG. 14C shows basal/squamous subtype versus CD8 T cell infiltration.
  • FIG. 15 shows ULBP2 expression of primary and non-primary site tumor sites in lung cancer from GSE248249.
  • FIGs. 16A-16E depict gene levels in five different datasets, each bifurcated by either HPV negative or HPV positive status of the patients.
  • FIG. 16A shows TCGA
  • FIG. 16B shows GSE65858
  • FIG. 16C shows GSE39366,
  • FIG. 16D shows GSE40774, and
  • FIG. 16E shows GSE6791.
  • FIGs. 17A-17D depict ULBP2 expression in either lung or bladder datasets. Samples are stratified by grade and stage of cancer and subtype (squamous or adeno). FIG. 17A shows GSE42127, FIG. 17B shows GSE14814, FIG. 17C shows TCGA lung squamous, and FIG. 17D shows TCGA bladder subtyped by molecular characterization to only basal/squamous as in FIG. 3.
  • FIGs. 18A-18H depict ULBP2 expression in either head and neck, lung, or bladder cancers. Pre/post treatment of check point inhibitors of paired or unpaired samples, with lung stratified by basal and squamous is shown.
  • FIG. 18A shows GSE179730 (head and neck)
  • FIG. 18B shows GSE195832 (head and neck)
  • FIGs. 18C and 18D show GES248249 (lung) with either paired or unpaired samples
  • FIGs. 18D and 18E show GSE248378 (lung) with either adeno or squamous breakout
  • FIG. 18G shows GSE248249 (lung) with outcome data from checkpoint inhibitors
  • FIG. 18H shows GSE212810 (bladder) with outcome data.
  • FIGs. 19A-19D depict ULBP2 expression from TCGA in patients that have EGFR mutations (FIG. 19A), highest and lowest quartile of EGFR expression (FIG. 19B), EGFR copy number alterations (FIG. 19C), and primary patients that have activating mutations that most likely will be prescribed the tyrosine kinase inhibitor (TKI) osimertinib (FIG. 19D).
  • TKI tyrosine kinase inhibitor
  • FIG. 20 is a graph showing ULBP2 expression in a bladder cancer dataset that stratifies patients into non-squamous disease and pure squamous cell carcinoma from GSE186691.
  • ULBP2 positive cancer such as basal squamous cancers, especially bladder cancer, display higher T cell infiltration (as seen by the expression of PDL1, PD1 and granzyme B). Without being bound by theory, together this suggests a high potential for efficacy of T cell engager therapies in this subset of patients.
  • a combination therapy of anti-ULBP2 antibody or antigen binding fragment thereof with a immune checkpoint inhibitor e.g., an anti-PD-1 antibody
  • a immune checkpoint inhibitor e.g., an anti-PD-1 antibody
  • the use of PD1 checkpoint inhibitors can further increase immune infiltration in subtypes where immune infiltration is lower, resulting in higher therapeutic potential.
  • Using specific chemotherapies or radiation therapy will induce ULBP2 in any subtype (even and especially in the adeno/luminal where it is lower). Therefore, a combination treatment and/or an anti-ULBP2 antibody or antigen binding fragment thereof treatment shortly following chemotherapy or radiation therapy will increase potency of the anti-ULBP2 antibody or antigen binding fragment thereof.
  • the disclosure provides a method for treating a cancer that expresses ULBP2.
  • the disclosure provides a method of decreasing a population of cancer cells in a subject.
  • the method of treating a cancer comprises administering an effective amount of an anti-ULBP2 antibody or antigen binding fragment thereof.
  • the method of treating a cancer comprises administering an effective amount of an anti-ULBP2 antibody or antigen binding fragment thereof and a chemotherapy agent, a radiation therapy, and/or an immune checkpoint inhibitor.
  • the cancer expresses UL16 binding protein 2 (ULBP2).
  • the cancer expresses KRT5 and/or SERPINB3.
  • the cancer is a squamous cancer (e.g., basal squamous cancer).
  • the basal squamous cancer is bladder cancer.
  • the basal squamous cancer is lung cancer.
  • the basal squamous cancer is esophageal cancer.
  • the basal squamous cancer is head and neck cancer.
  • the cancer is an adenocarcinoma (e.g., lung adenocarcinoma, pancreatic adenocarcinoma, bladder adenocarcinoma, esophageal adenocarcinoma, or colon adenocarcinoma).
  • the cancer is metastatic cancer.
  • anti-ULBP2 antibody is an antibody that binds specifically to ULBP2 antigen.
  • two types of anti-ULBP2 antibodies can be distinguished according to Cragg, M. S., et al., Blood 103 (2004) 2738-2743; and Cragg, M. S sharp et al., Blood 101 (2003) 1045-1052.
  • ULBP5 (RAET1G) encompasses naturally occurring variants of ULBP5, including, for example, splice variants or allelic variants.
  • ULBP5 includes, for example, human ULBP5 protein (UniProt ID: Q6H3X3), which is 334 amino acids in length.
  • the disclosure provides isolated antibodies that bind to ULBP5 (RAET1G).
  • the anti-ULBP5 antibody binds to a human ULBP5 polypeptide or a portion thereof.
  • the human ULBP5 polypeptide comprises the amino acid sequence of SEQ ID NO: 422.
  • ULBP6 (RAET1L) encompasses naturally occurring variants of ULBP6, including, for example, splice variants or allelic variants.
  • ULBP6 includes, for example, human ULBP6 protein (UniProt ID: Q5VY80), which is 246 amino acids in length.
  • the disclosure provides isolated antibodies that bind to ULBP6 (RAET1L).
  • the anti-ULBP6 antibody binds to a human ULBP6 polypeptide or a portion thereof.
  • the human ULBP6 polypeptide comprises the amino acid sequence of SEQ ID NO: 423.
  • antibodies that bind to anti-ULBP2/5/6 may be performed on the “E12” anti-ULBP2/5/6 antibody to produce affinity modulated anti-ULBP2/5/6 antibodies. Also provided herein are antibodies that bind to anti-ULBP2. In some embodiments, alanine scanning mutagenesis may be performed on the “A06” anti-ULBP2 antibody to produce affinity modulated anti-ULBP2 antibodies.
  • an anti-ULBP2/5/6 antibody of the disclosure comprises any one of the VH and VL sequences listed in Table 1.
  • Table 1 the underlined sequences are CDR sequence according to Kabat and the bolded sequences are CDR sequences according to Chothia.
  • an anti-ULBP2 antibody of the disclosure comprises: a) a heavy chain variable region (VH) comprising a VH complementarity determining region 1 (VHCDRI), a VH complementarity determining region 2 (VHCDR2) and a VH complementarity determining region 3 (VHCDRI); and b) a light chain variable region (VL) comprising a VL complementarity determining region 1 (VLCDRI), a VL complementarity determining region 2 (VLCDR2) and a VL complementarity determining region 3 (VLCDRI).
  • VH heavy chain variable region
  • VHCDRI VH complementarity determining region 1
  • VHCDR2 VHCDR2
  • VHCDRI VH complementarity determining region 3
  • VLCDRI light chain variable region
  • the disclosure provides an antibody (e.g. including antibody fragments, such as single chain variable fragments (scFvs) which specifically bind to ULBP2, wherein the antibody comprises a) a heavy chain variable region (VH) comprising a i) a VH complementarity determining region 1 (VHCDRI) comprising the amino acid sequence of SEQ ID NO: 5 or 6, ii) a VH complementarity determining region 2 (VHCDR2) comprising the amino acid sequence of SEQ ID NO: 7 or 8, iii) a VH complementarity determining region 3 (VHCDRS) comprising the amino acid sequence of SEQ ID NO: 9; and b) a light chain variable region (VL) comprising a i) a VL complementarity determining region 1 (VLCDRI) comprising the amino acid sequence of SEQ ID NO: 10, ii) a VL complementarity determining region 2 (VLCDR2) comprising the amino acid sequence of SEQ
  • Exemplary anti-ULBP2 antibodies of the disclosure include ULBP2-01, ULBP2-02, E12, A06.
  • the anti-ULBP2 antibody ULBP2-02 has a VH region comprising the amino acid sequence shown in SEQ ID NO: 4 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 3.
  • the anti-ULBP2 antibody E12 has a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 5, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 9; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 10, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 11, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 12.
  • the anti-ULBP2 antibody A06 has a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 428, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 430, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 432; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 433, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 434, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 435.
  • the anti-ULBP2 antibody A06 has a VH region comprising the amino acid sequence shown in SEQ ID NO: 427 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 426.
  • bispecific antibodies comprising a first antigen binding domain that binds a first antigen (e.g. cell surface antigen, CD3s) and a second antigen binding domain that binds to a second antigen (e.g. ULBP2/5/6).
  • a first antigen e.g. cell surface antigen, CD3s
  • a second antigen binding domain that binds to a second antigen (e.g. ULBP2/5/6).
  • the bispecific antibody has the following structure: a first heavy chain polypeptide (Hl) comprising a variable region (VH1), and a constant region (CHI) having a constant region 1 domain (CH1H1), a hinge region (H1H), a constant region 2 domain (CH1H2) and a constant region 3 domain (CH1H3); and a first light chain polypeptide (LI) comprising a variable region (VL1) and a constant region (CL1), and a second heavy chain polypeptide (H2) comprising a variable region (VH2), and a constant region (CH2) having a constant region 1 domain (CH2H1), a hinge region (H2H), a constant region 2 domain (CH2H2) and a constant region 3 domain (CH2H3); and second light chain polypeptide (L2) comprising a variable region (VL2) and a constant region (CL2).
  • Hl first heavy chain polypeptide
  • CHI constant region having a constant region 1 domain (CH1H1), a hinge
  • the bispecific antibody of the disclosure comprises a second antigen binding domain (e.g. binding to ULBP2/5/6) comprising any one of the VH2 and VL2 sequences listed in Table 1.
  • a second antigen binding domain e.g. binding to ULBP2/5/6
  • Table 1 the underlined sequences are CDR sequence according to Kabat and the bolded sequences are CDR sequences according to Chothia.
  • the bispecific antibody of the disclosure comprises a second antigen binding domain (e.g. binding to ULBP2) comprising: a) a heavy chain variable region (VH2) comprising a VH complementarity determining region 1 (VH2CDR1), a VH complementarity determining region 2 (VH2CDR2) and a VH2 complementarity determining region 3 (VH2CDR3); and b) a light chain variable region (VL2) comprising a VL complementarity determining region 1 (VL2CDR1), a VL complementarity determining region 2 (VL2CDR2) and a VL complementarity determining region 3 (VL2CDR3).
  • VH2CDR1 VH complementarity determining region 1
  • VH2CDR2CDR2CDR2CDR2 VH2 complementarity determining region 2
  • VL2CDR3 VL complementarity determining region 3
  • the bispecific antibody comprises any one of the anti- ULBP2/5/6 antibodies of the disclosure.
  • Exemplary anti-ULBP2/5/6 antibodies of the disclosure include ULBP2-01, ULBP2-02 and E12.
  • the bispecific antibody comprises any one of the anti-ULBP2 antibodies of the disclosure.
  • Exemplary anti- ULBP2 antibodies of the disclosure include A06.
  • the disclosure provides an isolated antibody (e.g. monospecific antibody or bispecific antibody) which specifically binds to ULPB2/5/6 and competes with any of the foregoing antibodies.
  • an isolated antibody e.g. monospecific antibody or bispecific antibody
  • the present disclosure provides an antibody (e.g. monospecific antibody or bispecific antibody) that binds to ULBP2/5/6 and competes with an antibody as described herein, including ULBP2-01, ULBP2-02, A06 and E12.
  • an antibody e.g. monospecific antibody or bispecific antibody
  • the disclosure also provides CDR portions of antibodies to ULBP2/5/6 antibodies based on CDR contact regions.
  • CDR contact regions are regions of an antibody that imbue specificity to the antibody for an antigen.
  • CDR contact regions include the residue positions in the CDRs and Vernier zones which are constrained in order to maintain proper loop structure for the antibody to bind a specific antigen. See, e.g., Makabe et al., J. Biol. Chem., 283:1156-1166, 2007. Determination of CDR contact regions is well within the skill of the art.
  • the binding affinity (KD) of the ULBP2/5/6 antibody e.g. monospecific antibody or bispecific antibody
  • ULBP2/5/6 such as human ULBP2 (e.g., (SEQ ID NO: 421), ULBP5 (SEQ ID NO: 422), ULBP6 (SEQ ID NO: 423)) can be about 0.001 to about 5000 nM.
  • the binding affinity is about any of 5000 nM, 4500 nM, 4000 nM, 3500 nM, 3000 nM, 2500 nM, 2000 nM, 1789 nM, 1583 nM, 1540 nM, 1500 nM, 1490 nM, 1064 nM, 1000 nM, 933 nM, 894 nM, 750 nM, 705 nM, 678 nM, 532 nM, 500 nM, 494 nM, 400 nM, 349 nM, 340 nM, 353 nM, 300 nM, 250 nM, 244 nM, 231 nM, 225 nM, 207 nM, 200 nM, 186 nM, 172 nM, 136 nM, 113 nM, 104 nM, 101 nM, 100 nM, 90 nM, 83 nM, 1064 nM,
  • the disclosure provides a nucleic acid encoding any of the foregoing isolated anti-ULBP2/5/6 antibodies (e.g. monospecific antibody or bispecific antibody).
  • the disclosure provides a vector comprising such a nucleic acid.
  • the disclosure provides a host cell comprising such a nucleic acid.
  • the disclosure provides anti-CD3 antibodies.
  • the anti-CD3 antibodies are multispecific (e.g., bispecific) and bind, in addition to CD3 or a fragment thereof, a second biological molecule (e.g., a cell surface antigen, e.g., a disease associated antigen ).
  • a second biological molecule e.g., a cell surface antigen, e.g., a disease associated antigen.
  • Antibodies of the disclosure are useful, for example, for treating or delaying the progression of a cell proliferative disorder (e.g., cancer) or an autoimmune disorder, or for enhancing immune function in a subject having such a disorder.
  • CD3 cluster of differentiation 3
  • mammals such as primates (e.g. humans, cynomolgus monkey) and rodents (e.g., mice and rats), unless otherwise indicated, including, for example, CD3s, CD3y, CD3a, and CD3P chains.
  • CD3 is a cell surface complex expressed on T cells in association with the T cell receptor. The CD3 complex is required for the activation of CD8+ and CD4+ T lymphocytes.
  • CD3 gamma chain one CD3 delta chain
  • CD3 epsilon chains which associate with each other to form a CD3 epsilon/gamma heterodimer, and a CD3 epsilon/delta heterodimer.
  • the two CD3 heterodimers, together with the T cell receptor (TCR) and the signal-transducing zeta chain homodimer form the T cell receptor complex.
  • CD3 encompasses “full-length” unprocessed CD3 (e.g., unprocessed or unmodified CD3s or CD3y), as well as any form of CD3 that results from processing in the cell.
  • the term also encompasses naturally occurring variants of CD3, including, for example, splice variants or allelic variants.
  • CD3 includes, for example, human CD3s protein (NCBI RefSeq No. NP_000724), which is 207 amino acids in length.
  • the disclosure provides isolated antibodies that bind to CD3. In some embodiments, the disclosure provides antibodies that bind to CD3s. In some instances the anti-CD3s antibody binds to a human CD3s polypeptide or a cynomolgus monkey (cyno) CD3s polypeptide. In some instances, the human CD3 polypeptide or the cyno CD3 polypeptide is a human CD3s polypeptide (SEQ ID NO: 419) or a cyno CD3s polypeptide (SEQ ID NO: 420), respectively.
  • the anti-CD3 antibody binds to an epitope within a fragment of CD3s (e.g., human CD3s) consisting of amino acid residues 1-26 or amino acid residues 1-27 of human CD3s (SEQ ID NO: 419).
  • a fragment of CD3s e.g., human CD3s
  • amino acid residues 1-26 or amino acid residues 1-27 of human CD3s SEQ ID NO: 419.
  • a useful method for identification of residues or regions of an antibody that may be targeted for mutagenesis is called “alanine scanning mutagenesis” as described by Cunningham and Wells (1989) Science, 244:1081-1085.
  • a residue or group of target residues e.g., charged residues such as Arg, Asp, His, Lys, and Glu
  • a neutral or negatively charged amino acid e.g., alanine or polyalanine
  • a crystal structure of an antigen-antibody complex is used to identify contact points between the antibody and antigen. Such contact residues and neighboring residues may be targeted or eliminated as candidates for substitution. Variants may be screened to determine whether they contain the desired properties.
  • anti-CD3s antibodies are provided herein.
  • alanine scanning mutagenesis was performed on the “SP34” anti-CD3s antibody to produce affinity modulated anti-CD3s antibodies of the disclosure.
  • an anti-CD3s antibody of the disclosure comprises any one of the VH and VL sequences listed in Table 4.
  • Table 4 the underlined sequences are CDR sequence according to Kabat and the bolded sequences are CDR sequences according to Chothia.
  • an anti-CD3 antibody of the disclosure comprises: a) a heavy chain variable region (VH) comprising a VH complementarity determining region 1 (VHCDRI), a VH complementarity determining region 2 (VHCDR2) and a VH complementarity determining region 3 (VHCDRI); and b) a light chain variable region (VL) comprising a VL complementarity determining region 1 (VLCDRI), a VL complementarity determining region 2 (VLCDR2) and a VL complementarity determining region 3 (VLCDRI).
  • VH heavy chain variable region
  • VHCDRI VH complementarity determining region 1
  • VHCDR2 VH complementarity determining region 2
  • VHCDRI VH complementarity determining region 3
  • VLCDRI light chain variable region
  • the disclosure provides an antibody (e.g. including antibody fragments, such as single chain variable fragments (scFvs) which specifically bind to CD3s, wherein the antibody comprises a) a heavy chain variable region (VH) comprising a i) a VH complementarity determining region 1 (VHCDRI) comprising the amino acid sequence of SEQ ID NO: 29, 30, 31, 32 or 33, ii) a VH complementarity determining region 2 (VHCDR2) comprising the amino acid sequence of SEQ ID NO: 34, 35 or 36, iii) a VH complementarity determining region 3 (VHCDRI) comprising the amino acid sequence of SEQ ID NO: 37, 38, 39, 40 or 41 ; and b) a light chain variable region (VL) comprising a i) i) a VL complementarity determining region 1 (VLCDRI) comprising the amino acid sequence of SEQ ID NO: 42, ii) a VL complement
  • the anti-CD3 antibody CD3-A1 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 46.
  • the anti-CD3 antibody CD3-A1 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 13 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 25.
  • the anti-CD3 antibody CD3-A2 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 44, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 45.
  • the anti-CD3 antibody CD3-A2 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 13 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 27.
  • the anti-CD3 antibody CD3-A3 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 45.
  • the anti-CD3 antibody CD3-A3 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 14 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 23.
  • the anti-CD3 antibody CD3-A4 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 38; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 47.
  • the anti-CD3 antibody CD3-A4 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 15 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 26.
  • the anti-CD3 antibody CD3-A5 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 39; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 47.
  • the anti-CD3 antibody CD3-A5 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 16 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 26.
  • the anti-CD3 antibody CD3-A6 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 30, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 45.
  • the anti-CD3 antibody CD3-A6 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 17 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 22.
  • the anti-CD3 antibody CD3-A7 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 35, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 38; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 45.
  • the anti-CD3 antibody CD3-A7 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 18 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 22.
  • the anti-CD3 antibody CD3-A8 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 35, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 38; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 47.
  • the anti-CD3 antibody CD3-A8 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 18 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 26.
  • the anti-CD3 antibody CD3-A9 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 40; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 47.
  • the anti-CD3 antibody CD3-A9 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 19 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 26.
  • the anti-CD3 antibody CD3-A10 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 41; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 47.
  • the anti-CD3 antibody CD3-A10 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 20 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 26.
  • the anti-CD3 antibody CD3-A11 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 31, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 37; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 45.
  • the anti-CD3 antibody CD3-A12 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 13 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 24.
  • the anti-CD3 antibody CD3-A13 comprises a VH region comprising a VHCDRI comprising the amino acid sequence of SEQ ID NO: 29, a VHCDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VHCDR3 comprising the amino acid sequence of SEQ ID NO: 39; and a VL region comprising a VLCDRI comprising the amino acid sequence of SEQ ID NO: 42, a VLCDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VLCDR3 comprising the amino acid sequence of SEQ ID NO: 48.
  • the anti-CD3 antibody CD3-A13 comprises a VH region comprising the amino acid sequence shown in SEQ ID NO: 16 and a VL region comprising the amino acid sequence shown in SEQ ID NO: 28.
  • bispecific antibodies comprising a first antigen binding domain that binds a first antigen (e.g. CD3s) and a second antigen binding domain that binds to a second antigen (e.g. disease associated antigen).
  • a first antigen e.g. CD3s
  • a second antigen binding domain that binds to a second antigen (e.g. disease associated antigen).
  • the bispecific antibody of the disclosure comprises a first antigen binding domain (e.g. binding to CD3) comprising any one of the VH1 and VL1 sequences listed in Table 4.
  • a first antigen binding domain e.g. binding to CD3
  • the underlined sequences are CDR sequence according to Kabat and the bolded sequences are CDR sequences according to Chothia.
  • the bispecific antibody comprises any one of the anti-CD3 antibodies of the disclosure.
  • Exemplary anti-CD3 antibodies of the disclosure include CD3- Al, CD3-A2, CD3-A3, CD3-A4, CD3-A5, CD3-A6, CD3-A7, CD3-A8, CD3-A9, CD3-A10, CD3-A11, CD3-A12 and CD3-A13.
  • the disclosure provides an isolated antibody (e.g. monospecific antibody or bispecific antibody) which specifically binds to CD3s and competes with any of the foregoing antibodies.
  • an isolated antibody e.g. monospecific antibody or bispecific antibody
  • the present disclosure provides an antibody (e.g. monospecific antibody or bispecific antibody) that binds to CD3s and competes with an antibody as described herein, including CD3-A1, CD3-A2, CD3-A3, CD3-A4, CD3-A5, CD3-A6, CD3- A7, CD3-A8, CD3-A9, CD3-A10, CD3-A11, CD3-A12 and CD3-A13.
  • an antibody e.g. monospecific antibody or bispecific antibody
  • the disclosure also provides CDR portions of antibodies to CD3s antibodies based on CDR contact regions.
  • CDR contact regions are regions of an antibody that imbue specificity to the antibody for an antigen.
  • CDR contact regions include the residue positions in the CDRs and Vernier zones which are constrained in order to maintain proper loop structure for the antibody to bind a specific antigen. See, e.g. , Makabe et al., J. Biol. Chem., 283:1156-1166, 2007. Determination of CDR contact regions is well within the skill of the art.
  • the binding affinity (KD) of the anti-CD3s antibodies of the disclosure e.g. monospecific antibody or bispecific antibody
  • human CD3s such as human CD3s (e.g., (SEQ ID NO: 419)
  • KD binding affinity
  • the binding affinity is about any of 5000 nM, 4500 nM, 4000 nM, 3500 nM, 3000 nM, 2500 nM, 2000 nM, 1789 nM, 1583 nM, 1540 nM, 1500 nM, 1490 nM, 1064 nM, 1000 nM, 933 nM, 894 nM, 750 nM, 705 nM, 678 nM, 532 nM, 500 nM, 494 nM, 400 nM, 349 nM, 340 nM, 353 nM, 300 nM, 250 nM, 244 nM, 231 nM, 225 nM, 207 nM, 200 nM, 186 nM, 172 nM, 136 nM, 113 nM, 104 nM, 101 nM, 100 nM, 90 nM, 83 nM, 1064 nM,
  • the binding affinity is less than about any of 5000 nM, 4000 nM, 3000 nM, 2000 nM, 1000 nM, 900 nM, 800 nM, 250 nM, 200 nM, 100 nM, 50 nM, 30 nM, 20 nM, 10 nM, 7.5 nM, 7 nM, 6.5 nM, 6 nM, 5 nM, 4.5 nM, 4 nM, 3.5 nM, 3 nM, 2.5 nM, 2 nM, 1.5 nM, 1 nM, or 0.5 nM.
  • the binding affinity (KD) of the anti- CD3s antibodies of the disclosure e.g. monospecific antibody or bispecific antibody
  • cynomolgus CD3s such as cynomolgus CD3s (e.g., (SEQ ID NO: 422)
  • SEQ ID NO: 422 cynomolgus CD3s
  • the binding affinity is about any of 5000 nM, 4500 nM, 4000 nM, 3500 nM, 3000 nM, 2500 nM, 2000 nM, 1789 nM, 1583 nM, 1540 nM, 1500 nM, 1490 nM, 1064 nM, 1000 nM, 933 nM, 894 nM, 750 nM, 705 nM, 678 nM, 532 nM, 500 nM, 494 nM, 400 nM, 349 nM, 340 nM, 353 nM, 300 nM, 250 nM, 244 nM, 231 nM, 225 nM, 207 nM, 200 nM, 186 nM, 172 nM, 136 nM, 113 nM, 104 nM, 101 nM, 100 nM, 90 nM, 83 nM, 1064 nM,
  • the binding affinity is less than about any of 5000 nM, 4000 nM, 3000 nM, 2000 nM, 1000 nM, 900 nM, 800 nM, 250 nM, 200 nM, 100 nM, 50 nM, 30 nM, 20 nM, 10 nM, 7.5 nM, 7 nM, 6.5 nM, 6 nM, 5 nM, 4.5 nM, 4 nM, 3.5 nM, 3 nM, 2.5 nM, 2 nM, 1.5 nM, 1 nM, or 0.5 nM.
  • the disclosure provides a nucleic acid encoding any of the foregoing isolated anti-CD3s antibodies (e.g. monospecific antibody or bispecific antibody).
  • the disclosure provides a vector comprising such a nucleic acid.
  • the disclosure provides a host cell comprising such a nucleic acid.
  • bispecific antibodies comprising a first antigen binding domain that binds a first antigen (e.g. CD3s) and a second antigen binding domain that binds to a second antigen (e.g. ULBP2/5/6).
  • a first antigen e.g. CD3s
  • a second antigen binding domain that binds to a second antigen (e.g. ULBP2/5/6).
  • the bispecific antibody has the following structure: a first heavy chain polypeptide (Hl) comprising a variable region (VH1), and a constant region (CHI) having a constant region 1 domain (CHIHI), a hinge region (H1H), a constant region 2 domain (CH1H2) and a constant region 3 domain (CH I ns); and a first light chain polypeptide (LI) comprising a variable region (VL1) and a constant region (CL1), and a second heavy chain polypeptide (H2) comprising a variable region (VH2), and a constant region (CH2) having a constant region 1 domain (CH2HI), a hinge region (H2H), a constant region 2 domain (CH2H2) and a constant region 3 domain (CH2H3); and second light chain polypeptide (L2) comprising a variable region (VL2) and a constant region (CL2).
  • Hl first heavy chain polypeptide
  • CHI constant region having a constant region 1 domain (CHIHI), a hinge region (H1
  • the bispecific antibody of the disclosure comprises a first antigen binding domain (e.g. binding to CD3s) comprising: a) a heavy chain variable region (VH1) comprising a VH complementarity determining region 1 (VHICDRI), a VH complementarity determining region 2 (VH1CDR2) and a VH complementarity determining region 3 (VHICDRS); and b) a light chain variable region (VL) comprising a VL complementarity determining region 1 (VLICDRI), a VL complementarity determining region 2 (VL1CDR2) and a VL complementarity determining region 3 (VLICDRS); and a second antigen binding domain (e.g. binding to CD3s) comprising: a) a heavy chain variable region (VH1) comprising a VH complementarity determining region 1 (VHICDRI), a VH complementarity determining region 2 (VH1CDR2) and a VH complementarity determining region 3 (VHICD
  • binding to ULBP2/5/6) comprising: a) a heavy chain variable region (VH2) comprising a VH complementarity determining region 1 (VH2CDRI), a VH complementarity determining region 2 (VH2CDR2) and a VH2 complementarity determining region 3 (VH2CDR3); and b) a light chain variable region (VL2) comprising a VL complementarity determining region 1 (VL2CDRI), a VL complementarity determining region 2 (VL2CDR2) and a VL complementarity determining region 3 (VL2CDR3).
  • Tables 5 and 6 provide exemplary CDR sequences of the anti-CD3s antibodies provided herein.
  • the bispecific antibody of the disclosure comprises a first antigen binding domain (e.g. binding to CD3s) comprising any one of the VH1 and VL1 sequences listed in Table 4 and a second antigen binding domain (e.g. binding to ULBP2/5/6) comprising any one of the VH2 and VL2 sequences listed in Table 1.
  • a first antigen binding domain e.g. binding to CD3s
  • a second antigen binding domain e.g. binding to ULBP2/5/6 comprising any one of the VH2 and VL2 sequences listed in Table 1.
  • the bispecific antibody of the disclosure comprises a first heavy chain polypeptide (Hl) and a first light chain polypeptide (LI); and a second heavy chain polypeptide (H2) and a second light chain polypeptide (L2) comprising any one of the sequences listed in Table 7 and Table 8.
  • the italicized sequences are the heavy chain variable regions and the light chain variable regions.
  • the underlined sequences are CDRs according to Kabat and the bolded sequences are CDRs according to Chothia.
  • the bispecific antibody of the disclosure provided herein comprises a Hl that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% identical to the amino acid sequence of the Hl sequences listed in Table 7 and Table 8.
  • the bispecific antibody of the disclosure provided herein comprises a LI that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% identical to the amino acid sequence of the LI sequences listed in Table 7 and Table 8.
  • the bispecific antibody of the disclosure provided herein comprises a H2 that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% identical to the amino acid sequence of the H2 sequences listed in Table 7 and Table 8.
  • the bispecific antibody of the disclosure provided herein comprises a L2 that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 100% identical to the amino acid sequence of the L2 sequences listed in Table 7 and Table 8.
  • the Hl amino acid sequence is numbered in accordance with SEQ ID NO: 439.
  • the LI amino acid sequence is numbered in accordance with SEQ ID NO: 438.
  • the H2 amino acid sequence is numbered in accordance with SEQ ID NO: 437.
  • the L2 amino acid sequence is numbered in accordance with SEQ ID NO: 436.
  • CD3s x ULBP2/5/6 bispecific antibodies of the disclosure include EIP0174, EIP0175, EIP0187, EIP0205, EIP0206, EIP0207, EIP0208, EIP0294, EIP0295, EIP0306, EIP0307, EIP0318, EIP0340, EIP0342, EIP0354, EIP0356, EIP0367, EIP0377, EIP0404, EIP0406, EIP0473, and EIP0598.
  • Bispecific antibodies comprise a first antigen binding domain that binds CD3s which comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a
  • Bispecific antibodies comprise a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, having VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a V
  • the bispecific antibody EIP0174 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acid at position 128 (EU numbering) of the CHIHI is a C and the amino acid at position 118 (EU numbering) of the CL1 is a C; the amino acid at position 220 (EU numbering) of the H1H is a S and the amino acid at position 214 (EU numbering) of the CL1 is an S; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering)
  • the bispecific antibody EIP0174 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering) of the CH2H3 is a C; the amino acid at position 366 (EU numbering) of the CH2H3 is a W; and the amino acid at position 447 (EU numbering) of the CH2H3 is deleted, when numbered in accordance with H2 amino acid sequence of SEQ NO: 437 and L2 amino acid sequence of SEQ NO
  • the bispecific antibody EIP0174 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0174 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 58, a LI comprising the amino acid sequence of SEQ ID NO: 57, a H2 comprising the amino acid sequence of SEQ ID NO: 56, and a L2 comprising the amino acid sequence of SEQ ID NO: 55.
  • the bispecific antibody EIP0175 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at position
  • the bispecific antibody EIP0175 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acid at position 134 (EU numbering) of the CH2HI is a C and the amino acid at position 116 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering)
  • the bispecific antibody EIP0175 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0175 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 62, a H2 comprising the amino acid sequence of SEQ ID NO: 60, a LI comprising the amino acid sequence of SEQ ID NO: 61, and a L2 comprising the amino acid sequence of SEQ ID NO: 59.
  • the bispecific antibody EIP0187 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at position
  • the bispecific antibody EIP0187 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acid at position 136 (EU numbering) of the CH2HI is a C and the amino acid at position 114 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering)
  • the bispecific antibody EIP0187 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0187 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 66, a H2 comprising the amino acid sequence of SEQ ID NO: 64, a LI comprising the amino acid sequence of SEQ ID NO: 65, and a L2 comprising the amino acid sequence of SEQ ID NO: 63.
  • the bispecific antibody EIP0205 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 147 (EU numbering) of the CHIHI is a K and the amino acid at position 131 (EU numbering) of the CL1 is a D; the amino acid at position 173 (EU numbering) of the CHIHI is a C and the amino acid at position 162 (EU numbering) of the CL1 is a C; the amino acid at position 220 (EU numbering) of the H1H is a S and the amino acid at position 214 (EU numbering) of the CL1 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering)
  • the amino acid at position 366 (EU numbering) of the CHlm is a S
  • the amino acid at position 368 (EU numbering) of the CHlm is a A
  • the amino acid at position 407 (EU numbering) of CH1H3 is a V
  • the amino acid at position 447 (EU numbering) of the CHlm is deleted, when numbered in accordance with Hl amino acid sequence of SEQ NO: 439 and LI amino acid sequence of SEQ NO: 438.
  • the bispecific antibody EIP0205 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering) of the CH2H3 is a C; the amino acid at position 366 (EU numbering) of the CH2H3 is a W; and the amino acid at position 447 (EU numbering) of the CH2H3 is deleted, when numbered in accordance with H2 amino acid sequence of SEQ NO: 437 and L2 amino acid sequence of SEQ NO
  • the bispecific antibody EIP0205 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0205 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 70, a H2 comprising the amino acid sequence of SEQ ID NO: 68, a LI comprising the amino acid sequence of SEQ ID NO: 69, and a L2 comprising the amino acid sequence of SEQ ID NO: 67.
  • the bispecific antibody EIP0206 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acid at position 173 (EU numbering) of the CHIHI is a C and the amino acid at position 162 (EU numbering) of the CL1 is a C; the amino acid at position 220 (EU numbering) in the H1H is a S and the amino acid at position 214 in CL1 is a S;
  • the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and [0173] the amino acid at position 447 (EU numbering) of the CH1H3 is deleted, when numbered in accordance with Hl amino acid sequence of SEQ NO: 439 and LI amino acid sequence of SEQ NO: 438.
  • the bispecific antibody EIP0206 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acid at position 134 (EU numbering) of the CH2HI is a C and the amino acid at position 116 (EU numbering) of the CL2 is a C; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering) of the CH2H3 is a C; the amino acid at position 366 (EU numbering) of the CH2H3 is a W; and the amino acid at position 447
  • the bispecific antibody EIP0206 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0206 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 74, a H2 comprising the amino acid sequence of SEQ ID NO: 72, a LI comprising the amino acid sequence of SEQ ID NO: 73, and a L2 comprising the amino acid sequence of SEQ ID NO: 71.
  • the bispecific antibody EIP0207 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at position
  • the bispecific antibody EIP0207 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acid at position 131 (EU numbering) of the CH2HI is a C and the amino acid at position 114 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering)
  • the bispecific antibody EIP0207 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0207 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 78, a H2 comprising the amino acid sequence of SEQ ID NO: 76, a LI comprising the amino acid sequence of SEQ ID NO: 77, and a L2 comprising the amino acid sequence of SEQ ID NO: 75.
  • the bispecific antibody EIP0208 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at position
  • the bispecific antibody EIP0208 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 170 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering) of the H2H are an A; the amino acid at position 354 (
  • the bispecific antibody EIP0208 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0208 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 82, a H2 comprising the amino acid sequence of SEQ ID NO: 80, a LI comprising the amino acid sequence of SEQ ID NO: 81, and a L2 comprising the amino acid sequence of SEQ ID NO: 79.
  • the bispecific antibody EIP0294 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHI HI is a E and the amino acid at position 137 (EU numbering) of the CL1 is a K and the amino acid at position 179 (EU numbering) of the CL1 is a E; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 40
  • amino acid at position 447 (EU numbering) of the CH1H3 is deleted, when numbered in accordance with Hl amino acid sequence of SEQ NO: 439 and LI amino acid sequence of SEQ NO: 438.
  • the bispecific antibody EIP0294 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 171 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU number of
  • the bispecific antibody EIP0294 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0294 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 86, a H2 comprising the amino acid sequence of SEQ ID NO: 84, a LI comprising the amino acid sequence of SEQ ID NO: 85, and a L2 comprising the amino acid sequence of SEQ ID NO: 83.
  • the bispecific antibody EIP0295 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at position
  • the bispecific antibody EIP0295 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 171 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU number of
  • the bispecific antibody EIP0295 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0295 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 90, a H2 comprising the amino acid sequence of SEQ ID NO: 88, a LI comprising the amino acid sequence of SEQ ID NO: 89, and a L2 comprising the amino acid sequence of SEQ ID NO: 87.
  • the bispecific antibody EIP0306 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHI HI is a E and the amino acid at position 137 (EU numbering) of the CL1 is a K and the amino acid at position 179 (EU numbering) of the CL1 is a E; the amino acid at position 185 (EU numbering) of the CHIHI is a E and the amino acid at position 179 (EU numbering) of the CL1 is a E; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU)
  • the amino acid at position 368 (EU numbering) of the CHlm is a A; the amino acid at position 407 (EU numbering) of CHlm is a V; and the amino acid at position 447 (EU numbering) of the CHlm is deleted, when numbered in accordance with Hl amino acid sequence of SEQ NO: 439 and LI amino acid sequence of SEQ NO: 438.
  • the bispecific antibody EIP0306 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acid at position 171 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering)
  • the bispecific antibody EIP0306 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0306 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 94, a H2 comprising the amino acid sequence of SEQ ID NO: 92, a LI comprising the amino acid sequence of SEQ ID NO: 93, and a L2 comprising the amino acid sequence of SEQ ID NO: 91.
  • the bispecific antibody EIP0307 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at position
  • the bispecific antibody EIP0307 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acid at position 171 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering)
  • the bispecific antibody EIP0307 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0307 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 98, a H2 comprising the amino acid sequence of SEQ ID NO: 96, a LI comprising the amino acid sequence of SEQ ID NO: 97, and a L2 comprising the amino acid sequence of SEQ ID NO: 95.
  • the bispecific antibody EIP0318 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 147 (EU numbering) of the CHIHI is a K and the amino acid at position 131 (EU numbering) of the CL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CHlm is a S; the amino acid at position 368 (EU
  • the bispecific antibody EIP0318 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acid at position 171 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering)
  • the bispecific antibody EIP0318 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0318 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 102, a H2 comprising the amino acid sequence of SEQ ID NO: 100, a LI comprising the amino acid sequence of SEQ ID NO: 101, and a L2 comprising the amino acid sequence of SEQ ID NO: 99.
  • the bispecific antibody EIP0340 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a D and the amino acid at position 38 (Kabat numbering) of the VL1 is a K; the amino acid at position 185 (EU numbering) of the CHI HI is a E and the amino acid at position 137 (EU numbering) of the CL1 is a K; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at
  • the bispecific antibody EIP0340 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a K and the amino acid at position 38 (Kabat numbering) of the VL2 is a D; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 136 (EU numbering) of the CH2HI is a C and the amino acid at position 114 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU number of
  • the bispecific antibody EIP0340 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 14, a VH2 comprising the amino acid sequence of SEQ ID NO: 4, a VL1 comprising the amino acid sequence of SEQ ID NO: 23, and a VL2 comprising the amino acid sequence of SEQ ID NO: 3.
  • the bispecific antibody EIP0340 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 106, a H2 comprising the amino acid sequence of SEQ ID NO: 104, a LI comprising the amino acid sequence of SEQ ID NO: 105, and a L2 comprising the amino acid sequence of SEQ ID NO: 103.
  • the bispecific antibody EIP0342 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a D and the amino acid at position 38 (Kabat numbering) of the VL1 is a K; the amino acid at position 185 (EU numbering) of the CHI HI is a E and the amino acid at position 137 (EU numbering) of the CL1 is a K; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at
  • the bispecific antibody EIP0342 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a K and the amino acid at position 38 (Kabat numbering) of the VL2 is a D; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 171 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU number
  • the bispecific antibody EIP0342 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 14, a VH2 comprising the amino acid sequence of SEQ ID NO: 4, a VL1 comprising the amino acid sequence of SEQ ID NO: 23, and a VL2 comprising the amino acid sequence of SEQ ID NO: 3.
  • the bispecific antibody EIP0342 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 110, a H2 comprising the amino acid sequence of SEQ ID NO: 108, a LI comprising the amino acid sequence of SEQ ID NO: 109, and a L2 comprising the amino acid sequence of SEQ ID NO: 107.
  • the bispecific antibody EIP0354 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a D and the amino acid at position 38 (Kabat numbering) of the VL1 is a K; the amino acid at position 185 (EU numbering) of the CHIHI is a K and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (EU numbering) of the CH1H3 is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at position 4
  • the bispecific antibody EIP0354 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a K and the amino acid at position 38 (Kabat numbering) of the VL2 is a D; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 171 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU number
  • the bispecific antibody EIP0354 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 14, a VH2 comprising the amino acid sequence of SEQ ID NO: 4, a VL1 comprising the amino acid sequence of SEQ ID NO: 23, and a VL2 comprising the amino acid sequence of SEQ ID NO: 3.
  • the bispecific antibody EIP0354 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 114, a H2 comprising the amino acid sequence of SEQ ID NO: 112, a LI comprising the amino acid sequence of SEQ ID NO: 113, and a L2 comprising the amino acid sequence of SEQ ID NO: 111.
  • the bispecific antibody EIP0356 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a D and the amino acid at position 38 (Kabat numbering) of the VL1 is a K; the amino acid at position 147 (EU numbering) of the CHIHI is a K and the amino acid at position 131 (EU numbering) of the CL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a E and the amino acid at position 137 (EU numbering) of the CL1 is a K; the amino acid at position 145 (EU numbering) of the CHIHI is a S and the amino acid at position 180 (EU numbering) of the CL1 is a E; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering)
  • the bispecific antibody EIP0356 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a K and the amino acid at position 38 (Kabat numbering) of the VL2 is a D; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 170 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU number
  • the bispecific antibody EIP0356 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 14, a VH2 comprising the amino acid sequence of SEQ ID NO: 4, a VL1 comprising the amino acid sequence of SEQ ID NO: 23, and a VL2 comprising the amino acid sequence of SEQ ID NO: 3.
  • the bispecific antibody EIP0356 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 118, a H2 comprising the amino acid sequence of SEQ ID NO: 116, a LI comprising the amino acid sequence of SEQ ID NO: 117, and a L2 comprising the amino acid sequence of SEQ ID NO: 115.
  • the bispecific antibody EIP0367 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 147 (EU numbering) of the CHIHI is a D and the amino acid at position 131 (EU numbering) of the CL1 is a K; the amino acid at position 185 (EU numbering) of the CHIHI is a E and the amino acid at position 137 (EU numbering) of the CL1 is a D; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (
  • the bispecific antibody EIP0367 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 137 (EU numbering) of the CL2 is a K and the amino acid at position 138 (EU numbering) of the CL2 is a R; the amino acid at position 170 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU number).
  • the bispecific antibody EIP0367 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0367 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 122, a H2 comprising the amino acid sequence of SEQ ID NO: 120, a LI comprising the amino acid sequence of SEQ ID NO: 121, and a L2 comprising the amino acid sequence of SEQ ID NO: 119.
  • the bispecific antibody EIP0377 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 147 (EU numbering) of the CHIHI is a D and the amino acid at position 145 (EU numbering) of the CHIHI is a S and the amino acid at position 131 (EU numbering) of the CL1 is a K; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CHlm is a C; the amino acid at position 366 (EU numbering) of the CHlm is a S; the amino acid at position 368 (EU numbering) of the CHlm is a A; the amino acid at position 407 (EU
  • amino acid at position 447 (EU numbering) of the CHlm is deleted, when numbered in accordance with Hl amino acid sequence of SEQ NO: 439 and LI amino acid sequence of SEQ NO: 438.
  • the bispecific antibody EIP0377 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 170 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering) of the H2H are an A; the amino acid at position 354 (
  • the bispecific antibody EIP0377 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0377 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 126, a H2 comprising the amino acid sequence of SEQ ID NO: 124, a LI comprising the amino acid sequence of SEQ ID NO: 125, and a L2 comprising the amino acid sequence of SEQ ID NO: 123.
  • the bispecific antibody EIP0404 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a D and the amino acid at position 38 (Kabat numbering) of the VL1 is a K; the amino acid at position 147 (EU numbering) of the CHIHI is a K and the amino acid at position 131 (EU numbering) of the CL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a E and the amino acid at position 137 (EU numbering) of the CL1 is a K; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CHlm is a S; the amino acid at position 368 (EU
  • the bispecific antibody EIP0404 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a K and the amino acid at position 38 (Kabat numbering) of the VL2 is a D; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acid at position 136 (EU numbering) of the CH2HI is a C and the amino acid at position 114 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering)
  • the bispecific antibody EIP0404 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 24, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0404 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 130, a H2 comprising the amino acid sequence of SEQ ID NO: 128, a LI comprising the amino acid sequence of SEQ ID NO: 129, and a L2 comprising the amino acid sequence of SEQ ID NO: 127.
  • the bispecific antibody EIP0406 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a E; the amino acid at position 147 (EU numbering) of the CHIHI is a K and the amino acid at position 131 (EU numbering) of the CL1 is a D; the amino acid at position 185 (EU numbering) of the CHIHI is a E and the amino acid at position 137 (EU numbering) of the CL1 is a K; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CH1H3 is a S; the amino acid at position 368 (
  • the bispecific antibody EIP0406 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 136 (EU numbering) of the CH2HI is a C and the amino acid at position 114 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU number of
  • the bispecific antibody EIP0406 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 24, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0406 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 134, a H2 comprising the amino acid sequence of SEQ ID NO: 132, a LI comprising the amino acid sequence of SEQ ID NO: 133, and a L2 comprising the amino acid sequence of SEQ ID NO: 131.
  • the bispecific antibody EIP0473 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 147 (EU numbering) of the CHIHI is a D and the amino acid at position 131 (EU numbering) of the CL1 is a K; the amino acid at position 185 (EU numbering) of the CHIHI is a D and the amino acid at position 137 (EU numbering) of the CL1 is a K; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CHlm is a S; the amino acid at position 368 (EU
  • the bispecific antibody EIP0473 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 187 (EU numbering) of the CH2HI is a D and the amino acid at position 138 (EU numbering) of the CL2 is a K; the amino acid at position 171 (EU numbering) of the CH2HI is a C and the amino acid at position 162 (EU numbering) of the CL2 is a C; the amino acid at position 220 (EU numbering) in the H2H is a S and the amino acid at position 214 (EU numbering) of the CL2 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU number of
  • the bispecific antibody EIP0473 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 4, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 3.
  • the bispecific antibody EIP0473 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 138, a H2 comprising the amino acid sequence of SEQ ID NO: 136, a LI comprising the amino acid sequence of SEQ ID NO: 137, and a L2 comprising the amino acid sequence of SEQ ID NO: 135.
  • the bispecific antibody EIP0598 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 170 (EU numbering) of the CHI HI is a S and the amino acid at position 131 (EU numbering) of the CL1 is a D; the amino acid at position 173 (EU numbering) of the CHIHI is a C and the amino acid at position 162 (EU numbering) of the CL1 is a C; the amino acid at position 220 (EU numbering) of the H1H is a S and the amino acid at position 214 (EU numbering) of the CL1 is an S; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering)
  • the bispecific antibody EIP0598 comprises the following amino acids substitutions in the H2 and L2: the amino acid at position 39 (Kabat numbering) of the VH2 is a D and the amino acid at position 38 (Kabat numbering) of the VL2 is a K; the amino acid at position 147 (EU numbering) of the CH2HI is a D and the amino acid at position 180 (EU numbering) of the CL2 is a R; the amino acids at positions 234, 235, and 237 (EU numbering) of the H2H are an A; the amino acid at position 354 (EU numbering) of the CH2H3 is a C; the amino acid at position 366 (EU numbering) of the CH2H3 is a W; and the amino acid at position 447 (EU numbering) of the CH2H3 is deleted, when numbered in accordance with H2 amino acid sequence of SEQ NO: 437 and L2 amino acid sequence of SEQ NO
  • the bispecific antibody EIP0598 comprises a VH1 comprising the amino acid sequence of SEQ ID NO: 13, a VH2 comprising the amino acid sequence of SEQ ID NO: 2, a VL1 comprising the amino acid sequence of SEQ ID NO: 22, and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0598 comprises a Hl comprising the amino acid sequence of SEQ ID NO: 142, a H2 comprising the amino acid sequence of SEQ ID NO: 140, a LI comprising the amino acid sequence of SEQ ID NO: 141, and a L2 comprising the amino acid sequence of SEQ ID NO: 139.
  • any one of the bispecific antibodies shown above in Table 7 can be further modified by substituting any one of the anti-CD3s antigen binding regions with any one of the anti- CD3s binding regions shown in Tables 7-9.
  • the anti-CD3s antigen binding regions of bispecific antibody “EIP0205” can be substituted with any one of the anti-CD3s binding regions shown in Tables 7-9 to produce the bispecific antibodies of the disclosure.
  • Exemplary antibodies are shown in Table 8.
  • the underlined sequences are CDR sequence according to Kabat and the bolded sequences are CDR sequences according to Chothia.
  • exemplary, CD3s x ULBP2/5/6 bispecific antibodies of the disclosure include EIP0527, EIP0624, EIP0486, EIP0626, EIP0483, EIP0623, EIP0621, EIP0625, EIP0622, EIP0525.
  • Bispecific antibodies EIP0527, EIP0624, EIP0486, EIP0626, EIP0483, EIP0623, EIP0621, EIP0625, EIP0622, and EIP0525 comprise the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D;
  • the amino acid at position 147 (EU numbering) of the CHIHI is a K and the amino acid at position 131 (EU numbering) of the CL1 is a D; the amino acid at position 173 (EU numbering) of the CHIHI is a C and the amino acid at position 162 (EU numbering) of the CL1 is a C; the amino acid at position 220 (EU numbering) of the H1H is a S and the amino acid at position 214 (EU numbering) of the CL1 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C; the amino acid at position 366 (EU numbering) of the CHlm is a S; the amino acid at position 368 (EU numbering) of the CHlm is a A; the amino acid at position 407 (EU numbering) of CH1H3 is a V; and the amino acid at position
  • Bispecific antibodies EIP0527, EIP0624, EIP0486, EIP0626, EIP0483, EIP0623, EIP0621, EIP0625, EIP0622, and EIP0525 have a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • Bispecific antibodies EIP0527, EIP0624, EIP0486, EIP0626, EIP0483, EIP0623, EIP0621, EIP0625, EIP0622, and EIP0525 comprise a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • Bispecific antibodies EIP0527, EIP0624, EIP0486, EIP0626, EIP0483, EIP0623, EIP0621, EIP0625, EIP0622, and EIP0525 have a H2 comprising the amino acid sequence of SEQ ID NO: 68; and a L2 comprising the amino acid sequence of SEQ ID NO: 67.
  • the bispecific antibody EIP0527 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 46.
  • the bispecific antibody EIP0527 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 25. In some embodiments, the bispecific antibody EIP0527 comprises a Hl having the amino acid sequence of SEQ ID NO: 146 and a LI having the amino acid sequence of SEQ ID NO: 145.
  • the bispecific antibody EIP0624 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0624 comprises a VH1 having the amino acid sequence of SEQ ID NO: 15 and a VL1 having the amino acid sequence of SEQ ID NO: 26. In some embodiments, the bispecific antibody EIP0624 comprises a Hl having the amino acid sequence of SEQ ID NO: 150 and a LI having the amino acid sequence of SEQ ID NO: 149.
  • the bispecific antibody EIP0486 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 44; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0626 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 39; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0626 comprises a VH1 having the amino acid sequence of SEQ ID NO: 16 and a VL1 having the amino acid sequence of SEQ ID NO: 26. In some embodiments, the bispecific antibody EIP0626 comprises a Hl having the amino acid sequence of SEQ ID NO: 158 and a LI having the amino acid sequence of SEQ ID NO: 157.
  • the bispecific antibody EIP0483 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 30; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0483 comprises a VH1 having the amino acid sequence of SEQ ID NO: 17 and a VL1 having the amino acid sequence of SEQ ID NO: 22. In some embodiments, the bispecific antibody EIP0483 comprises a Hl having the amino acid sequence of SEQ ID NO: 162 and a LI having the amino acid sequence of SEQ ID NO: 161.
  • the bispecific antibody EIP0623 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 35; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0623 comprises a VH1 having the amino acid sequence of SEQ ID NO: 18 and a VL1 having the amino acid sequence of SEQ ID NO: 26. In some embodiments, the bispecific antibody EIP0623 comprises a Hl having the amino acid sequence of SEQ ID NO: 166 and a LI having the amino acid sequence of SEQ ID NO: 165.
  • the bispecific antibody EIP0621 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 40; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0625 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 35; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0625 comprises a VH1 having the amino acid sequence of SEQ ID NO: 18 and a VL1 having the amino acid sequence of SEQ ID NO: 22. In some embodiments, the bispecific antibody EIP0625 comprises a Hl having the amino acid sequence of SEQ ID NO: 174 and a LI having the amino acid sequence of SEQ ID NO: 173.
  • the bispecific antibody EIP0622 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 41; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0622 comprises a VH1 having the amino acid sequence of SEQ ID NO: 20 and a VL1 having the amino acid sequence of SEQ ID NO: 26. In some embodiments, the bispecific antibody EIP0622 comprises a Hl having the amino acid sequence of SEQ ID NO: 178 and a LI having the amino acid sequence of SEQ ID NO: 177.
  • the bispecific antibody EIP0525 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 31; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0525 comprises a VH1 having the amino acid sequence of SEQ ID NO: 21 and a VL1 having the amino acid sequence of SEQ ID NO: 22. In some embodiments, the bispecific antibody EIP0525 comprises a Hl having the amino acid sequence of SEQ ID NO: 182 and a LI having the amino acid sequence of SEQ ID NO: 181.
  • exemplary, CD3s x ULBP2/5/6 bispecific antibodies of the disclosure include EIP0630, EIP0540, EIP0628, EIP0542, EIP0627, EIP0515, EIP0477, EIP0541, EIP0513, EIP0629.
  • bispecific antibodies EIP0630, EIP0540, EIP0628, EIP0542, EIP0627, EIP0515, EIP0477, EIP0541, EIP0513, and EIP0629 have the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 170 (EU numbering) of the CHIHI is a S and the amino acid at position 131 (EU numbering) of the CL1 is a D; the amino acid at position 173 (EU numbering) of the CHIHI is a C and the amino acid at position 162 (EU numbering) of the CL1 is a C; the amino acid at position 220 (EU numbering) of the H1H is a S and the amino acid at position 214 (EU numbering) of the CL1 is an S; the amino acids at positions 234, 235,
  • Bispecific antibodies EIP0630, EIP0540, EIP0628, EIP0542, EIP0627, EIP0515, EIP0477, EIP0541, EIP0513, and EIP0629 have a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, having VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 comprising the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, having VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • Bispecific antibodies EIP0630, EIP0540, EIP0628, EIP0542, EIP0627, EIP0515, EIP0477, EIP0541, EIP0513, and EIP0629 have a VH2 having the amino acid sequence of SEQ ID NO: 2; and a VL2 having the amino acid sequence of SEQ ID NO: 1.
  • the bispecific antibody EIP0542 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 39; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0542 comprises a VH1 having the amino acid sequence of SEQ ID NO: 16 and a VL1 having the amino acid sequence of SEQ ID NO: 26. In some embodiments, the bispecific antibody EIP0542 comprises a Hl having the amino acid sequence of SEQ ID NO: 198 and a LI having the amino acid sequence of SEQ ID NO: 197.
  • the bispecific antibody EIP0627 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 30; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0627 comprises a VH1 having the amino acid sequence of SEQ ID NO: 17 and a VL1 having the amino acid sequence of SEQ ID NO: 22. In some embodiments, the bispecific antibody EIP0627 comprises a Hl having the amino acid sequence of SEQ ID NO: 202 and a LI having the amino acid sequence of SEQ ID NO: 201.
  • the bispecific antibody EIP0515 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 35; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0515 comprises a VH1 having the amino acid sequence of SEQ ID NO: 18 and a VL1 having the amino acid sequence of SEQ ID NO: 26. In some embodiments, the bispecific antibody EIP0515 comprises a Hl having the amino acid sequence of SEQ ID NO: 206 and a LI having the amino acid sequence of SEQ ID NO: 205.
  • the bispecific antibody EIP0477 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 40; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0477 comprises a VH1 having the amino acid sequence of SEQ ID NO: 19 and a VL1 having the amino acid sequence of SEQ ID NO: 26. In some embodiments, the bispecific antibody EIP0477 comprises a Hl having the amino acid sequence of SEQ ID NO: 210 and a LI having the amino acid sequence of SEQ ID NO: 209.
  • the bispecific antibody EIP0541 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 35; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0541 comprises a VH1 having the amino acid sequence of SEQ ID NO: 18 and a VL1 having the amino acid sequence of SEQ ID NO: 22. In some embodiments, the bispecific antibody EIP0541 comprises a Hl having the amino acid sequence of SEQ ID NO: 214 and a LI having the amino acid sequence of SEQ ID NO: 213.
  • the bispecific antibody EIP0513 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 41; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0513 comprises a VH1 having the amino acid sequence of SEQ ID NO: 20 and a VL1 having the amino acid sequence of SEQ ID NO: 26. In some embodiments, the bispecific antibody EIP0513 comprises a Hl having the amino acid sequence of SEQ ID NO: 218 and a LI having the amino acid sequence of SEQ ID NO: 217.
  • the bispecific antibody EIP0629 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 31; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0629 comprises a VH1 having the amino acid sequence of SEQ ID NO: 21 and a VL1 having the amino acid sequence of SEQ ID NO: 22. In some embodiments, the bispecific antibody EIP0629 comprises a Hl having the amino acid sequence of SEQ ID NO: 222 and a LI having the amino acid sequence of SEQ ID NO: 221.
  • exemplary CD3s x ULBP2/5/6 bispecific antibodies of the present disclosure comprise an amino acid at position 446 (EU numbering) and/or at position 447 (EU numbering) of the CH1H3 and/or of the CH2H3.
  • the amino acid at position 446 (EU numbering) is a G.
  • the exemplary bispecific antibodies comprise an amino acid at position 446 (EU numbering) of the CHlm and/or of the CH2H3, the bispecific antibodies further comprise an amino acid at position 447 (EU numbering) of the CHlm and/or of the CH2H3.
  • the amino acid at position 447 (EU numbering) is a K.
  • exemplary CD3s x ULBP2/5/6 bispecific antibodies of the present disclosure comprise an amino acid at position 446 (EU numbering) of the CH1H3 or CH2H3.
  • the bispecific antibodies comprise an amino acid at position 446 (EU numbering) of the CHI H3 and CH2H3. In some embodiments, the bispecific antibodies comprise an amino acid at position 446 (EU numbering) of the CH1H3. In some embodiments, the bispecific antibodies comprise an amino acid at position 446 (EU numbering) of the CH2H3.
  • the amino acid at position 446 (EU numbering) of the CH1H3 and/or CH2H3 is a G. In some embodiments, wherein the bispecific antibodies comprise an amino acid at position 446 (EU numbering) of the CH1H3, the amino acid at position 446 (EU numbering) is a G. In some embodiments, wherein the bispecific antibodies comprise an amino acid at position 446 (EU numbering) of the CH2H3, the amino acid at position 446 (EU numbering) is a G. In some embodiments, wherein the bispecific antibodies comprise an amino acid at position 446 (EU numbering) of the CH1H3 and CH2H3, the amino acid at position 446 (EU numbering) of the CH1H3 and CH2H3 is a G.
  • exemplary CD3s x ULBP2/5/6 bispecific antibodies of the disclosure comprise an amino acid at position 447 (EU numbering) of the CH1H3 and/or of the CH2H3.
  • the bispecific antibodies comprise an amino acid at position 447 (EU numbering) of the CH1H3 or of the CH2H3.
  • the bispecific antibodies comprise an amino acid at position 447 (EU numbering) of the CH1H3 and of the CH2H3.
  • the bispecific antibodies comprise an amino acid at position 447 (EU numbering) of the CH1H3.
  • the bispecific antibodies comprise an amino acid at position 447 (EU numbering) of the CH2H3.
  • CD3s x ULBP2/5/6 bispecific antibodies comprise an amino acid at position 447 (EU numbering) of the CH1H3 and/or of the CH2H3, the amino acid at position 447 (EU numbering) is a K. In some embodiments, wherein the bispecific antibodies comprise an amino acid at position 447 (EU numbering) of the CH1H3, the amino acid at position 447 (EU numbering) is a K. In some embodiments, wherein the bispecific antibodies comprise an amino acid at position 447 (EU numbering) of the CH2H3, the amino acid at position 447 (EU numbering) is a K. Fusion Peptides
  • an antibody e.g. monospecific antibody or bispecific antibody
  • a fusion peptide fused to the N-terminus or the C-terminus of the first heavy chain polypeptide or the second heavy chain polypeptide.
  • T cell activation Critical to the initial T cell response is the capacity for T cells to detect foreign and mutated proteins through their T cell receptor.
  • This response often referred to as signal 1 of T cell activation, occurs when the T cell receptor engages a cell that displays a foreign or mutated protein fragment or antigen in a specific protein complex called the Major Histocompatibility Complex I (MHCI).
  • MHCI Major Histocompatibility Complex I
  • the activation of the T cell receptor is by itself both activating and auto-regulatory to T cells. Strong binding of the TCR to an MHCI complex creates chronic activation of the TCR.
  • This form of signal is associated with T cells that are reactive to self-antigens. T cells are programed to inactivate when they experience this form activation. T cells with TCR that bind weaker, but sufficient for activation, experience acute signaling with the potential to remain active and differentiate into memory T cells. This is emerging as important consideration in the design the T cell therapeutics.
  • T cell cytokine activation is important in T cell transitions, either from non-dividing to a state of rapid cell division or from one phenotypic state to another.
  • T cell cytokine receptors bind to cytokines that are produced by immune and non- immune cells and depending on the cytokine and the state of the T cell at the time of receiving the cytokine signal can induce cell proliferation, can sustain vitality, or can induce differentiation of T cells into a specialized cell state appropriate for sustained activation or inactivation following infection.
  • cytokines which can induce naive T cells to proliferate and promote T cell differentiation into memory T cells.
  • cytokines include but are not limited to IL-2, IL-7, IL- 10, IL- 12, IL- 15, IL- 18 and IL-21.
  • Costimulatory receptor activation referred to as signal 2 provides a context specific cell-to-cell reinforcement of T activation.
  • the most recognized form of costimulation occurs when T cells interact with activated antigen presenting cells through the T cell costimulatory receptor CD28 with CD80 and CD86 ligands found on APCs. These interactions can “prime” specific T cells armed with T cell receptors responsive to pathogen or cancer proteins.
  • costimulation induced at the site of infection and malignancies This includes costimulation that acts through CD2 and NKG2D receptors responsive to ligands like CD58 and ULI 6 binding proteins (e.g. ULBP2/5/6) that are induced in immune cells and epithelial cells upon viral infection. These signals provide not only reinforcement of T activation, but confirmation that the T cell’s lethal effector activities are targeted with single cell accuracy. While many costimulatory receptors have been discovered, the importance of each receptor’s specific context and the impact of concurrent signaling of multiple costimulatory receptors remains largely unknown and an area to greatly advance our understanding of T cell biology and creating possibilities for novel tumor-targeted T cell therapeutic development.
  • Costimulatory ligands include but are not limited to CD48, CD58, CD86, TNFSF9, OX40L, 4-1BBL, GITL, CD70, CD80, MR1, TNFSF4, ICOSL or ICOSLG.
  • CD58 is advantageous over other costimulatory ligands in that it is the primary costimulatory pathway available at the tumor site as tumor infiltrating T lymphocytes often lose expression of other costimulatory receptors like CD28, or due to the low immunogenicity of tumor cells, tumor cells do not sufficiently activate T cell, thus limiting the potential of inducible costimulatory receptors like 41BB.
  • the anti-CD3s antibodies of the disclosure induce varying levels of T cell receptor activation that confer alteration in T cell vitality and cytokine production. Accordingly, a fusion of the costimulatory ligand CD58 to the anti-CD3s bispecific antibody provides integrated costimulatory T cell activation for optimal T cell activation.
  • the bispecific antibody has a peptide fused to the N-terminus of the first heavy chain polypeptide (Hl). In some embodiments, the bispecific antibody has a peptide fused to the C-terminus of the first heavy chain polypeptide (Hl). In some embodiments, the bispecific antibody has a polypeptide fused to the N-terminus of the second heavy chain polypeptide (H2). In some embodiments, the bispecific antibody has a peptide fused to the C-terminus of the second heavy chain polypeptide (H2). Exemplary peptides include but are not limited to IL-2, IL-7, IL- 10, IL- 12, IL- 15, IL- 18, IL-21 or portions thereof.
  • Exemplary peptides include but are not limited to CD48, CD58, CD86, TNFSF9, OX40E, 4- 1BBE, GITE, CD70, CD80, MR1, TNFSF4, ICOSE, ICOSLG or portions thereof.
  • Exemplary peptide sequences that are fused to the bispecific antibodies include but are not limited to those listed in Table 9 and Table 10.
  • polypeptide is fused directly to the bispecific antibody.
  • polypeptide is fused indirectly through a linker.
  • the bispecific antibody fused with a peptide comprises a linker sequence.
  • Exemplary linker sequences include but are not limited to those listed in Table 11 and Table 12.
  • Bispecific antibody comprising a fusion peptide
  • the present disclosure provides a bispecific antibody (e.g., a bispecific antibody targeting ULBP2 and CD3) that is fused to a peptide (e.g., a costimulatory CD58 peptide described herein).
  • a bispecific antibody described e.g., a bispecific antibody targeting ULBP2 and CD3 herein comprises a peptide (e.g., a costimulatory CD58 peptide described herein) fused to the C-terminal of the heavy chain of the CD3 targeting arm.
  • a bispecific antibody described (e.g., a bispecific antibody targeting ULBP2 and CD3) herein comprises a peptide (e.g., a costimulatory CD58 peptide described herein) fused to the N-terminal of the heavy chain of the CD3 targeting arm.
  • a bispecific antibody described (e.g., a bispecific antibody targeting ULBP2 and CD3) herein comprises a peptide (e.g., a costimulatory CD58 peptide described herein) fused to the C-terminal of the light chain of the CD3 targeting arm.
  • a bispecific antibody described (e.g., a bispecific antibody targeting ULBP2 and CD3) herein comprises a peptide (e.g., a costimulatory CD58 peptide described herein) fused to the N-terminal of the light chain of the CD3 targeting arm.
  • a bispecific antibody described (e.g., a bispecific antibody targeting ULBP2 and CD3) herein comprises a peptide (e.g., a costimulatory CD58 peptide described herein) fused to the C-terminal of the heavy chain of the ULBP2 targeting arm.
  • a bispecific antibody described (e.g., a bispecific antibody targeting ULBP2 and CD3) herein comprises a peptide (e.g., a costimulatory CD58 peptide described herein) fused to the N-terminal of the heavy chain of the ULBP2 targeting arm.
  • a bispecific antibody described (e.g., a bispecific antibody targeting ULBP2 and CD3) herein comprises a peptide (e.g., a costimulatory CD58 peptide described herein) fused to the C-terminal of the light chain of the ULBP2 targeting arm.
  • a bispecific antibody described e.g., a bispecific antibody targeting ULBP2 and CD3 herein comprises a peptide (e.g., a costimulatory CD58 peptide described herein) fused to the N-terminal of the light chain of the ULBP2 targeting arm.
  • Table 13 sets forth exemplary bispecific antibodies targeting UBLP2 and CD3 that are fused to a CD58 peptide.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0820.
  • bispecific antibody EIP0820 comprises the following amino acids substitutions in the Hl and LI: the amino acid at position 39 (Kabat numbering) of the VH1 is a K and the amino acid at position 38 (Kabat numbering) of the VL1 is a D; the amino acid at position 147 (EU numbering) of the CHIHI is a K and the amino acid at position 131 (EU numbering) of the CL1 is a D; the amino acid at position 173 (EU numbering) of the CHIHI is a C and the amino acid at position 162 (EU numbering) of the CL1 is a C; the amino acid at position 220 (EU numbering) of the H1H is a S and the amino acid at position 214 (EU numbering) of the CL1 is a S; the amino acids at positions 234, 235, and 237 (EU numbering) in the H1H are an A; the amino acid at position 349 (EU numbering) of the CH1H3 is a C;
  • bispecific antibody EIP0820 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0820 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • bispecific antibody EIP0820 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 629; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • bispecific antibody EIP0820 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 621; and a L2 comprising the amino acid sequence of SEQ ID NO: 67.
  • the bispecific antibody EIP0820 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 30; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0820 comprises a VH1 having the amino acid sequence of SEQ ID NO: 17 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0820 comprises a Hl having the amino acid sequence of SEQ ID NO: 622 and a LI having the amino acid sequence of SEQ ID NO: 69.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0359.
  • bispecific antibody EIP0359 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0359 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • bispecific antibody EIP0359 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 224; and a L2 comprising the amino acid sequence of SEQ ID NO: 223.
  • the bispecific antibody EIP0359 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 32; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 36; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0359 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0359 comprises a Hl having the amino acid sequence of SEQ ID NO: 226 and a LI having the amino acid sequence of SEQ ID NO: 225.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0566.
  • bispecific antibody EIP0566 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0566 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0566 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 228; and a L2 comprising the amino acid sequence of SEQ ID NO: 227.
  • the bispecific antibody EIP0566 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 32; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 36; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0566 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 25.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0566 comprises a Hl having the amino acid sequence of SEQ ID NO: 230 and a LI having the amino acid sequence of SEQ ID NO: 229.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0634.
  • bispecific antibody EIP0634 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0634 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0634 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 232; and a L2 comprising the amino acid sequence of SEQ ID NO: 231.
  • the bispecific antibody EIP0634 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 32; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 36; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • bispecific antibody EIP0562 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0562 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0562 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 236; and a L2 comprising the amino acid sequence of SEQ ID NO: 235.
  • the bispecific antibody EIP0562 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 32; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 36; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 44; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0562 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 27.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0562 comprises a Hl having the amino acid sequence of SEQ ID NO: 238 and a LI having the amino acid sequence of SEQ ID NO: 237.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0636.
  • bispecific antibody EIP0636 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0636 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0636 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 240; and a L2 comprising the amino acid sequence of SEQ ID NO: 239.
  • the bispecific antibody EIP0636 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 32; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 36; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 39; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0636 comprises a VH1 having the amino acid sequence of SEQ ID NO: 16 and a VL1 having the amino acid sequence of SEQ ID NO: 26.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0636 comprises a Hl having the amino acid sequence of SEQ ID NO: 242 and a LI having the amino acid sequence of SEQ ID NO: 241.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0561.
  • bispecific antibody EIP0561 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0561 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO. 1. In some embodiments, bispecific antibody EIP0561 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 244; and a L2 comprising the amino acid sequence of SEQ ID NO: 243.
  • the bispecific antibody EIP0561 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 30; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0561 comprises a VH1 having the amino acid sequence of SEQ ID NO: 17 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0561 comprises a Hl having the amino acid sequence of SEQ ID NO: 246 and a LI having the amino acid sequence of SEQ ID NO: 245.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0633.
  • bispecific antibody EIP0633 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0633 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0633 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 248; and a L2 comprising the amino acid sequence of SEQ ID NO: 247.
  • the bispecific antibody EIP0633 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 35; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0633 comprises a VH1 having the amino acid sequence of SEQ ID NO: 18 and a VL1 having the amino acid sequence of SEQ ID NO: 26.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0633 comprises a Hl having the amino acid sequence of SEQ ID NO: 250 and a LI having the amino acid sequence of SEQ ID NO: 249.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0631.
  • bispecific antibody EIP0631 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0631 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0631 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 252; and a L2 comprising the amino acid sequence of SEQ ID NO: 251.
  • the bispecific antibody EIP0631 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 40; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0631 comprises a VH1 having the amino acid sequence of SEQ ID NO: 19 and a VL1 having the amino acid sequence of SEQ ID NO: 26.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0631 comprises a Hl having the amino acid sequence of SEQ ID NO: 254 and a LI having the amino acid sequence of SEQ ID NO: 253.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0635.
  • bispecific antibody EIP0635 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0635 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0635 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 256; and a L2 comprising the amino acid sequence of SEQ ID NO: 255.
  • the bispecific antibody EIP0635 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 35; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0635 comprises a VH1 having the amino acid sequence of SEQ ID NO: 18 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0635 comprises a Hl having the amino acid sequence of SEQ ID NO: 258 and a LI having the amino acid sequence of SEQ ID NO: 257.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0632.
  • bispecific antibody EIP0632 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0632 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0632 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 260; and a L2 comprising the amino acid sequence of SEQ ID NO: 259.
  • the bispecific antibody EIP0632 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 41; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0632 comprises a VH1 having the amino acid sequence of SEQ ID NO: 20 and a VL1 having the amino acid sequence of SEQ ID NO: 26.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0632 comprises a Hl having the amino acid sequence of SEQ ID NO: 262 and a LI having the amino acid sequence of SEQ ID NO: 261.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0565.
  • bispecific antibody EIP0565 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0565 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0565 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 264; and a L2 comprising the amino acid sequence of SEQ ID NO: 263.
  • the bispecific antibody EIP0565 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 31; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • bispecific antibody EIP0599 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0599 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0599 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 267; and a L2 comprising the amino acid sequence of SEQ ID NO: 330.
  • the bispecific antibody EIP0599 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 32; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 36; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • bispecific antibody EIP0640 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0640 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0640 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 271; and a L2 comprising the amino acid sequence of SEQ ID NO: 270,
  • the bispecific antibody EIP0640 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 46.
  • the bispecific antibody EIP0640 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 25.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0640 comprises a Hl having the amino acid sequence of SEQ ID NO: 273 and a LI having the amino acid sequence of SEQ ID NO: 272.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0567.
  • bispecific antibody EIP0567 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0567 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0567 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 275; and a L2 comprising the amino acid sequence of SEQ ID NO: 274,
  • the bispecific antibody EIP0567 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0567 comprises a VH1 having the amino acid sequence of SEQ ID NO: 15 and a VL1 having the amino acid sequence of SEQ ID NO: 26.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0567 comprises a Hl having the amino acid sequence of SEQ ID NO: 277 and a LI having the amino acid sequence of SEQ ID NO: 276.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0638.
  • bispecific antibody EIP0638 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0638 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0638 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 279; and a L2 comprising the amino acid sequence of SEQ ID NO: 278.
  • the bispecific antibody EIP0638 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 44; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0638 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 27.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0638 comprises a Hl having the amino acid sequence of SEQ ID NO: 281 and a LI having the amino acid sequence of SEQ ID NO: 280.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0568.
  • bispecific antibody EIP0568 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0568 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0568 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 283; and a L2 comprising the amino acid sequence of SEQ ID NO: 282.
  • the bispecific antibody EIP0568 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 39; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0568 comprises a VH1 having the amino acid sequence of SEQ ID NO: 16 and a VL1 having the amino acid sequence of SEQ ID NO: 26.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0568 comprises a Hl having the amino acid sequence of SEQ ID NO: 285 and a LI having the amino acid sequence of SEQ ID NO: 284.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0637.
  • the bispecific antibody EIP0564 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 35; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 38; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • bispecific antibody EIP0560 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0560 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO. 1. In some embodiments, bispecific antibody EIP0560 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 295; and a L2 comprising the amino acid sequence of SEQ ID NO: 294.
  • the bispecific antibody EIP0560 comprises a VH1 having the amino acid sequence of SEQ ID NO: 19 and a VL1 having the amino acid sequence of SEQ ID NO: 26.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0560 comprises a Hl having the amino acid sequence of SEQ ID NO: 297 and a LI having the amino acid sequence of SEQ ID NO: 296.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0662.
  • bispecific antibody EIP0662 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0662 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0662 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 299; and a L2 comprising the amino acid sequence of SEQ ID NO: 298.
  • the bispecific antibody EIP0662 comprises a VH1 having the amino acid sequence of SEQ ID NO: 18 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0662 comprises a Hl having the amino acid sequence of SEQ ID NO: 301 and a LI having the amino acid sequence of SEQ ID NO: 300.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0563.
  • bispecific antibody EIP0563 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • the bispecific antibody EIP0563 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 41; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 47.
  • the bispecific antibody EIP0563 comprises a VH1 having the amino acid sequence of SEQ ID NO: 20 and a VL1 having the amino acid sequence of SEQ ID NO: 26.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0563 comprises a Hl having the amino acid sequence of SEQ ID NO: 305 and a LI having the amino acid sequence of SEQ ID NO: 304.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0639.
  • bispecific antibody EIP0639 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0639 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0639 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 307; and a L2 comprising the amino acid sequence of SEQ ID NO: 306.
  • the bispecific antibody EIP0639 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 31; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0639 comprises a VH1 having the amino acid sequence of SEQ ID NO: 21 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0639 comprises a Hl having the amino acid sequence of SEQ ID NO: 309 and a LI having the amino acid sequence of SEQ ID NO: 308.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0547.
  • bispecific antibody EIP0547 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0547 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0547 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 311; and a L2 comprising the amino acid sequence of SEQ ID NO: 310.
  • the bispecific antibody EIP0547 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0547 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0547 comprises a Hl having the amino acid sequence of SEQ ID NO: 313 and a LI having the amino acid sequence of SEQ ID NO: 312.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0641.
  • bispecific antibody EIP0641 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0641 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0641 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 315; and a L2 comprising the amino acid sequence of SEQ ID NO: 314.
  • the bispecific antibody EIP0641 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 49 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0641 comprises a Hl having the amino acid sequence of SEQ ID NO: 317 and a LI having the amino acid sequence of SEQ ID NO: 316.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0360.
  • bispecific antibody EIP0360 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0360 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, bispecific antibody EIP0360 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 319; and a L2 comprising the amino acid sequence of SEQ ID NO: 318.
  • the bispecific antibody EIP0360 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 29; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 34; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0360 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a CD58 peptide comprising the amino acid sequence of SEQ ID NO: 50 is fused to the C-terminal of the heavy chain targeting CD3 (Hl).
  • the bispecific antibody EIP0360 comprises a Hl having the amino acid sequence of SEQ ID NO: 321 and a LI having the amino acid sequence of SEQ ID NO: 320.
  • an exemplary CD3s x ULBP2/5/6 bispecific antibody of the disclosure includes EIP0363.
  • bispecific antibody EIP0363 has a second antigen binding domain that binds ULBP2/5/6 comprising a VH2, comprising VH2CDRI having the amino acid sequence of SEQ ID NO: 5; a VH2CDR2 having the amino acid sequence of SEQ ID NO: 7; and a VH2CDR3 having the amino acid sequence of SEQ ID NO: 9; and a VL2, comprising a VL2CDRI having the amino acid sequence of SEQ ID NO: 10; a VL2CDR2 having the amino acid sequence of SEQ ID NO: 11; and a VL2CDR3 having the amino acid sequence of SEQ ID NO: 12.
  • bispecific antibody EIP0363 comprises a VH2 comprising the amino acid sequence of SEQ ID NO: 2; and a VL2 comprising the amino acid sequence of SEQ ID NO: 1.
  • bispecific antibody EIP0363 comprises a H2 comprising the amino acid sequence of SEQ ID NO: 323; and a L2 comprising the amino acid sequence of SEQ ID NO: 322.
  • the bispecific antibody EIP0363 comprises a first antigen binding domain that binds CD3s comprising a VH1, comprising a VHICDRI having the amino acid sequence of SEQ ID NO: 32; a VH1CDR2 having the amino acid sequence of SEQ ID NO: 36; and a VH1CDR3 having the amino acid sequence of SEQ ID NO: 37; and a VL1, comprising a VLICDRI having the amino acid sequence of SEQ ID NO: 42; a VL1CDR2 having the amino acid sequence of SEQ ID NO: 43; and a VL1CDR3 having the amino acid sequence of SEQ ID NO: 45.
  • the bispecific antibody EIP0363 comprises a VH1 having the amino acid sequence of SEQ ID NO: 13 and a VL1 having the amino acid sequence of SEQ ID NO: 22.
  • a IL-7 peptide is fused to the C-terminal of the heavy chain targeting CD3 (Hl). Accordingly, in some embodiments, the bispecific antibody EIP0363 comprises a Hl having the amino acid sequence of SEQ ID NO: 325 and a LI having the amino acid sequence of SEQ ID NO: 324.
  • the disclosure provides for the use of antibodies of the disclosure in the manufacture or preparation of a medicament.
  • the medicament is for treatment of a cell proliferative disorder (e.g., cancer, such as squamous cancer, adenocarcinoma, or metastatic cancer).
  • the medicament is for use in a method of treating a cell proliferative disorder or an autoimmune disorder comprising administering to an individual having a cell proliferative disorder or an autoimmune disorder an effective amount of the medicament.
  • the method further comprises administering to the individual an effective amount of at least one additional therapeutic agent, for example, as described below.
  • the medicament is for activating effector cells (e.g., T cells, e.g., CD8+ and/or CD4+ T cells), expanding (increasing) an effector cell population, reducing a target cell (e.g., a cell expressing ULBP2) population, and/or killing target cells (e.g., target tumor cells) in the individual.
  • effector cells e.g., T cells, e.g., CD8+ and/or CD4+ T cells
  • expanding (increasing) an effector cell population reducing a target cell (e.g., a cell expressing ULBP2) population
  • target cells e.g., target tumor cells
  • the medicament is for use in a method of enhancing immune function in an individual having a cell proliferative disorder or an autoimmune disorder comprising administering to the individual an amount effective of the medicament to activate effector cells (e.g., T cells, e.g., CD8+ and/or CD4+ T cells), expand (increase) an effector cell population, reduce a target cell (e.g. , a cell expressing ULBP2) population, and/or kill a target cell (e.g., target tumor cell).
  • effector cells e.g., T cells, e.g., CD8+ and/or CD4+ T cells
  • expand (increase) an effector cell population e.g., a cell expressing ULBP2) population
  • kill a target cell e.g., target tumor cell.
  • An "individual" according to any of the above embodiments may be a human.
  • the disclosure provides a method for treating a cell proliferative disorder (e.g., cancer).
  • the method comprises administering to the individual an effective amount of a bispecific antibody to activate effector cells (e.g., T cells, e.g., CD8+ and/or CD4+ T cells), expand (increase) an effector cell population, reduce a target cell (e.g., a cell expressing ULBP2) population, and/or kill a target cell (e.g., target tumor cell).
  • effector cells e.g., T cells, e.g., CD8+ and/or CD4+ T cells
  • expand (increase) an effector cell population e.g., a cell expressing ULBP2) population
  • kill a target cell e.g., target tumor cell
  • the method further comprises comparing a level of ULBP2 expression in the cancer sample to a level of ULBP2 expression in healthy /control/reference tissue to determine if the level of ULBP2 expression in the cancer sample is higher. If the level of ULBP2 expression is higher in the cancer sample, the subject receives a ULBP2 targeting therapeutic.
  • the ULBP2 targeting therapeutic is one or more of the antibodies (e.g., a bispecific antibody targeting ULBP2 and CD3) provided herein.
  • any method for measuring the level of ULBP2 can be employed. In some embodiments, this is any amount greater than a negative amount in a staining assay.
  • this is any amount above a level present in surrounding healthy/control/reference tissue or corresponding tissue from a healthy/control/reference subject.
  • high levels of expression of ULBP2 is defined in comparison to an expression level of ULBP2 in a non-cancer sample.
  • the expression level of ULBP2 in the cancer sample is at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 125%, 150%, 175%, 200%, 225%, 250%, 275%, 300%, 325%, 350%, 375%, 400%, 425%, 450%, 475% or 500% higher than the expression level of ULBP2 in the non-cancer sample.
  • the healthy /control/reference sample is a sample from a normal tissue.
  • the normal tissue is a tissue that is an adjacent tissue to the cancer in the subject.
  • the levels of the protein are above those understood to be “negative” to one of skill in the art for the particular assay.
  • Exemplary assays include but are not limited to RNA based assays, FACS, and IHC, each with their appropriate levels of positive and negative results. Any method of detecting the level of a protein in a sample is contemplated. One skilled in the art can select a suitable method depending on the type of sample being analyzed and the identity and number of proteins being detected.
  • Nonlimiting exemplary such methods include immunohistochemistry, ELISA, Western blotting, multiplex analyte detection (using, for example, Luminex technology), mass spectrometry, etc.
  • any method of detecting the level of an mRNA in a sample is contemplated.
  • One skilled in the art can select a suitable method depending on the type of sample being analyzed and the identity and number of mRNAs being detected.
  • Nonlimiting exemplary such methods include RT-PCR, quantitative RT-PCR and microarray-based methods, etc.
  • a ULBP2-positive cancer may be a basal squamous bladder cancer or lung cancer.
  • an ULBP2-positive cancer is a cancer that receives an anti-ULBP2 immunohistochemistry (IHC) or in situ hybridization (ISH) score greater than “0,” which corresponds to very weak or no staining in >90% of tumor cells.
  • a ULBP2-positive cancer expresses ULBP2 at a 1+, 2+ or 3+ level.
  • a ULBP2-positive cancer is a cancer that expresses ULBP2 according to a reverse-transcriptase PCR (RT-PCR) assay that detects ULBP2 mRNA.
  • the RT-PCR is quantitative RT-PCR.
  • the disclosure provides pharmaceutical formulations comprising any of the bispecific antibodies provided herein, e.g., for use in any of the above therapeutic methods.
  • a pharmaceutical formulation comprises any of the bispecific antibodies provided herein and a pharmaceutically acceptable carrier.
  • a pharmaceutical formulation comprises any of the bispecific antibodies provided herein and at least one additional therapeutic agent, for example, as described herein.
  • the disclosure provides a method wherein the additional therapeutic agent is a glucocorticoid.
  • the glucocorticoid is dexamethasone.
  • the additional therapeutic agent is a checkpoint inhibitor.
  • the term “inhibition” or “inhibitor” includes a reduction in a certain parameter, e.g., an activity, of a given molecule, e.g., an immune checkpoint inhibitor.
  • inhibition of an activity e.g., an activity of, e.g., PD-1, PD-L1, CTLA-4, TIM-3, CEACAM (e.g., CEACAM-1, CEACAM-3 and/or CEACAM-5), LAG-3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4 or TGFR beta, of at least 5%, 10%, 20%, 30%, 40% or more is included by this term.
  • the level of inhibition need not be 100%.
  • the checkpoint inhibitor is a PD-1 inhibitor.
  • the PD- 1 inhibitor is an anti-PDl antibody.
  • the PD-1 inhibitor is an anti PD-1 monoclonal antibody.
  • Exemplary anti-PD-1 monoclonal antibodies include, but are not limited to cemiplimab (Libtayo), nivolumab (Opdivo), pembrolizumab (Keytruda).
  • the checkpoint inhibitor is a PD-L1 inhibitor.
  • Exemplary PD-L1 inhibitors include but are not limited to avelumab (Bavencio), durvalumab (Imfinzi) and atezolizumab (Tecentriq).
  • Such combination therapies noted above encompass combined administration (where two or more therapeutic agents are included in the same or separate formulations), and separate administration, in which case, administration of the antibody of the disclosure can occur prior to, simultaneously, and/or following, administration of the additional therapeutic agent or agents.
  • administration of the bispecific antibody and administration of an additional therapeutic agent occur within about one month, or within about one, two or three weeks, or within about one, two, three, four, five, or six days, of each other.
  • Bispecific antibodies of the disclosure can also be used in combination with radiation therapy.
  • the additional therapeutic agent is a chimeric antigen receptor (CAR) T cell therapy.
  • the CAR-T cell therapy specifically binds CD 19.
  • Exemplary CAR-T cell therapies that specifically bind CD 19 include but are not limited to BREYANZI® (lisocabtagene maraleucel), TECARTUSTM (brexucabtagene autoleucel), KYMRIAHTM (tisagenlecleucel), YESCARTATM (axicabtagene ciloleucel), ABECMA® (idecabtagene vicleucel), or CARVYKTITM (ciltacabtagene autoleucel).
  • An antibody of the disclosure can be administered by any suitable means, including parenteral, intrapulmonary, and intranasal, and, if desired for local treatment, intralesional administration.
  • Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
  • the antibody is administered by subcutaneous administration.
  • an anti-CD3s antibody administered by subcutaneous injection exhibits a less toxic response in a patient than the same anti-CD3s antibody administered by intravenous injection.
  • Dosing can be by any suitable route, for example, by injections, such as intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic.
  • Various dosing schedules including but not limited to single or multiple administrations over various time-points, bolus administration, and pulse infusion are contemplated herein.
  • Antibodies of the disclosure would be formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners.
  • the antibody need not be, but is optionally formulated with one or more agents currently used to prevent or treat the disorder in question. The effective amount of such other agents depends on the amount of antibody present in the formulation, the type of disorder or treatment, and other factors discussed above. These are generally used in the same dosages and with administration routes as described herein, or about from 1 to 99% of the dosages described herein, or in any dosage and by any route that is empirically/clinically determined to be appropriate.
  • the therapeutically effective amount of the bispecific antibody administered to human will be in the range of about 0.01 to about 100 mg/kg of patient body weight whether by one or more administrations.
  • the antibody used is about 0.01 to about 45 mg/kg, about 0.01 to about 40 mg kg, about 0.01 to about 35 mg/kg, about 0.01 to about 30 mg/kg, about 0.01 to about 25 mg/kg, about 0.01 to about 20 mg/kg, about 0.01 to about 15 mg kg, about 0.01 to about 10 mg/kg, about 0.01 to about 5 mg/kg, or about 0.01 to about 1 mg/kg administered daily, for example.
  • a bispecific antibody described herein is administered to a human at a dose of about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1100 mg, about 1200 mg, about 1300 mg or about 1400 mg on day 1 of 21 -day cycles.
  • the dose may be administered as a single dose or as multiple doses (e.g., 2 or 3 doses), such as infusions.
  • the treatment would generally be sustained until a desired suppression of disease symptoms occurs.
  • One exemplary dosage of the antibody would be in the range from about 0.05 mg/kg to about 10 mg/kg.
  • one or more doses of about 0.5 mg/kg, 2.0 mg/kg, 4.0 mg kg, or 10 mg/kg (or any combination thereof) may be administered to the patient.
  • Such doses may be administered intermittently, for example, every week or every three weeks (e.g., such that the patient receives from about two to about twenty, or, for example, about six doses of the bispecific antibody).
  • An initial higher loading dose, followed by one or more lower doses may be administered. The progress of this therapy is easily monitored by conventional techniques and assays.
  • the methods may further comprise an additional therapy.
  • the additional therapy may be radiation therapy, surgery, chemotherapy, gene therapy, DMA therapy, viral therapy, RNA therapy, immunotherapy, bone marrow transplantation, nanotherapy, monoclonal antibody therapy, or a combination of the foregoing.
  • the additional therapy may be in the form of adjuvant or neoadjuvant therapy.
  • the additional therapy is the administration of small molecule enzymatic inhibitor or anti- metastatic agent.
  • the additional therapy is the administration of sideeffect limiting agents (e.g., agents intended to lessen the occurrence and/or severity of side effects of treatment, such as anti-nausea agents, etc.).
  • the additional therapy is radiation therapy.
  • the additional therapy is surgery. In some embodiments, the additional therapy is a combination of radiation therapy and surgery. In some embodiments, the additional therapy is gamma irradiation. In some embodiments, the additional therapy may be a separate administration of one or more of the therapeutic agents described above.
  • formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic) containing vesicles (such as LipofectinTM), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. Any of the foregoing mixtures may be appropriate in treatments and therapies in accordance with the present disclosure, provided that the active ingredient in the formulation is not inactivated by the formulation and the formulation is physiologically compatible and tolerable with the route of administration.
  • Therapeutic formulations of the disclosure are used to treat or alleviate a symptom associated with a cancer, such as, by way of non-limiting example, leukemias, lymphomas, breast cancer, colon cancer, ovarian cancer, bladder cancer, prostate cancer, glioma, lung & bronchial cancer, colorectal cancer, pancreatic cancer, esophageal cancer, liver cancer, urinary bladder cancer, kidney and renal pelvis cancer, oral cavity & pharynx cancer, uterine corpus cancer, and/or melanoma
  • a therapeutic regimen is carried out by identifying a subject, e.g. , a human patient suffering from (or at risk of developing) a cancer, using standard methods.
  • Efficaciousness of treatment is determined in association with any known method for diagnosing or treating the particular immune-related disorder. Alleviation of one or more symptoms of the immune-related disorder indicates that the antibody confers a clinical benefit.
  • Methods for the screening of antibodies that possess the desired specificity include, but are not limited to, enzyme linked immunosorbent assay (ELISA) and other immunologically mediated techniques known within the art.
  • ELISA enzyme linked immunosorbent assay
  • Antibodies directed against a target such as CD3s, ULBP2, or a combination thereof (or a fragment thereof), may be used in methods known within the art relating to the localization and/or quantitation of these targets, e.g., for use in measuring levels of these targets within appropriate physiological samples, for use in diagnostic methods, for use in imaging the protein, and the like).
  • antibodies specific any of these targets, or derivative, fragment, analog or homolog thereof, that contain the antibody derived antigen binding domain are utilized as pharmacologically active compounds (referred to hereinafter as “Therapeutics”).
  • An antibody of the disclosure can be used to isolate a particular target using standard techniques, such as immunoaffinity, chromatography or immunoprecipitation.
  • Antibodies of the disclosure (or a fragment thereof) can be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, e.g. , to determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling (z.e., physically linking) the antibody to a detectable substance.
  • detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials.
  • suitable enzymes include horseradish peroxidase, alkaline phosphatase, P-galactosidase, or acetylcholinesterase;
  • suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin;
  • suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin;
  • an example of a luminescent material includes luminol;
  • examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include 125 I, 131 1, 35 S or 3 H.
  • Antibodies of the disclosure may be used as therapeutic agents. Such agents will generally be employed to treat or prevent a disease or pathology associated with aberrant expression or activation of a given target in a subject.
  • An antibody preparation preferably one having high specificity and high affinity for its target antigen, is administered to the subject and will generally have an effect due to its binding with the target.
  • Administration of the antibody may abrogate or inhibit or interfere with the signaling function of the target.
  • Administration of the antibody may abrogate or inhibit or interfere with the binding of the target with an endogenous ligand to which it naturally binds.
  • a therapeutically effective amount of an antibody of the disclosure relates generally to the amount needed to achieve a therapeutic objective. As noted above, this may be a binding interaction between the antibody and its target antigen that, in certain cases, interferes with the functioning of the target.
  • the amount required to be administered will furthermore depend on the binding affinity of the antibody for its specific antigen, and will also depend on the rate at which an administered antibody is depleted from the free volume other subject to which it is administered.
  • Common ranges for therapeutically effective dosing of an antibody or antibody fragment of the disclosure may be, by way of nonlimiting example, from about 0.1 mg/kg body weight to about 50 mg/kg body weight. Common dosing frequencies may range, for example, from twice daily to once a week.
  • Antibodies or a fragment thereof of the disclosure can be administered for the treatment of a variety of diseases and disorders in the form of pharmaceutical compositions. Principles and considerations involved in preparing such compositions, as well as guidance in the choice of components are provided, for example, in Remington: The Science And Practice Of Pharmacy 19th ed. (Alfonso R. Gennaro, et al., editors) Mack Pub. Co., Easton, Pa.: 1995; Drug Absorption Enhancement: Concepts, Possibilities, Limitations, And Trends, Harwood Academic Publishers, Langhorne, Pa., 1994; and Peptide And Protein Drug Delivery (Advances In Parenteral Sciences, Vol. 4), 1991, M. Dekker, New York.
  • antibody fragments are used, the smallest inhibitory fragment that specifically binds to the binding domain of the target protein is preferred.
  • peptide molecules can be designed that retain the ability to bind the target protein sequence.
  • Such peptides can be synthesized chemically and/or produced by recombinant DNA technology. (See, e.g., Marasco et al., Proc. Natl. Acad. Sci. USA, 90: 7889-7893 (1993)).
  • the formulation can also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other.
  • the composition can comprise an agent that enhances its function, such as, for example, a cytotoxic agent, cytokine, chemotherapeutic agent, or growth-inhibitory agent.
  • an agent that enhances its function such as, for example, a cytotoxic agent, cytokine, chemotherapeutic agent, or growth-inhibitory agent.
  • Such molecules are suitably present in combination in amounts that are effective for the purpose intended.
  • the active ingredients can also be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacrylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles, and nanocapsules) or in macroemulsions.
  • colloidal drug delivery systems for example, liposomes, albumin microspheres, microemulsions, nano-particles, and nanocapsules
  • formulations to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes.
  • sustained-release preparations can be prepared. Suitable examples of sustained- release preparations include semipermeable matrices of solid hydrophobic polymers containing the antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No.
  • copolymers of L-glutamic acid and y ethyl-L-glutamate non-degradable ethylene-vinyl acetate
  • degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT TM (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate)
  • poly-D-(-)-3-hydroxybutyric acid While polymers such as ethylenevinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods.
  • Examples of indirect labeling include detection of a primary antibody using a fluorescently-labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently-labeled streptavidin.
  • bio sample is intended to include tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. Included within the usage of the term “biological sample”, therefore, is blood and a fraction or component of blood including blood serum, blood plasma, or lymph. That is, the detection method of the disclosure can be used to detect an analyte mRNA, protein, or genomic DNA in a biological sample in vitro as well as in vivo.
  • in vitro techniques for detection of an analyte mRNA include Northern hybridizations and in situ hybridizations.
  • In vitro techniques for detection of an analyte protein include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations, and immunofluorescence.
  • In vitro techniques for detection of an analyte genomic DNA include Southern hybridizations. Procedures for conducting immunoassays are described, for example in “ELISA: Theory and Practice: Methods in Molecular Biology”, Vol. 42, J. R. Crowther (Ed.) Human Press, Totowa, NJ, 1995; “Immunoassay”, E. Diamandis and T.
  • in vivo techniques for detection of an analyte protein include introducing into a subject a labeled anti- analyte protein antibody.
  • the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.
  • Some embodiments of this disclosure provide a method of treating a squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein in combination with a chemotherapy, a radiotherapy, and/or an immune checkpoint inhibitor.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3
  • Any suitable chemotherapy can be used in the methods described herein, including but not limited to a topoisomerase inhibitor, an anthracycline doxorubicin, a DNA break inducing antibiotic, a pyrimidine antagonist, or a platinum alkylating agent.
  • Any suitable radiotherapy can be used in the methods described herein, including but not limited to 3D conformal radiation therapy (3DCRT), image-guided radiation therapy (IGRT), intensity modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), brachytherapy, intraoperative radiation therapy (IORT), stereotactic radiosurgery (SRS), proton therapy, MRI linear accelerator, and/or stereotactic body radiation therapy (SBRT).
  • 3DCRT 3D conformal radiation therapy
  • IGRT image-guided radiation therapy
  • IMRT intensity modulated radiation therapy
  • VMAT volumetric modulated arc therapy
  • IORT intraoperative radiation therapy
  • SRS stereotactic radiosurgery
  • proton therapy MRI linear accelerator
  • SBRT stereotactic body radiation therapy
  • Any suitable immune checkpoint inhibitor can be used in the methods described herein, including but not limited to avelumab, durvalumab, atezolizumab, pembrolizumab, nivolumab, atezolizum
  • SCCs Squamous cell cancers
  • SCCs are malignancies caused by squamous cells, a subtype of epithelial cell found in the skin and mucous membranes. SCCs are commonly associated with skin cancer, but can develop in any organ lined by squamous cells, including but not limited to the bladder, lungs, esophagus, cervix, anus, vulva, penis, thymus, thyroid, and mouth (e.g., in head and neck squamous cell cancer, or oropharyngeal squamous cell cancer).
  • the disclosure provides a method of treating a bladder squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3
  • the disclosure provides a method of treating a lung squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • the disclosure provides a method of treating an esophageal squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3
  • the disclosure provides a method of treating a head and neck squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • the disclosure provides a method of treating a cervical squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3
  • the disclosure provides a method of treating a skin squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • the disclosure provides a method of treating a thymic squamous cell cancer in a subject, the method comprising administering to the subject an anti- ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3
  • the disclosure provides a method of treating an anal squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • the disclosure provides a method of treating a vulva squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3
  • the disclosure provides a method of treating a penile squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • the disclosure provides a method of treating an adrenocortical squamous cell cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3
  • the disclosure provides a method of treating a squamous cell cancer in a subject, the method comprising administering an anti-ULBP2 antibody provided herein and a chemotherapy agent, a radiation therapy, and/or an immune checkpoint inhibitor.
  • this disclosure provides a method of treating an adeno carcinoma in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein in combination with a chemotherapy, a radio therapy, and/or an immune checkpoint inhibitor.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3
  • the adenocarcinoma is lung adenocarcinoma, pancreatic adenocarcinoma, esophageal adenocarcinoma, bladder adenocarcinoma, or colon adenocarcinoma.
  • a subject having cancer has increases level of ULBP2 expression level in the cancer cells relative to a non-cancerous cell.
  • ULBP2 expression level is well correlated with the portion of squamous cell cancer in the cancer tissue.
  • an adenocarcinoma has increases ULBP2 expression when the cancer has an EGFR activating mutation or an EGFR copy number alteration (e.g., increased EGFP copy numbers).
  • EGRF activating mutations are generally know, for example as described by Gazdar, Oncogene. 2009 Aug; 28(Suppl 1): S24-S31.
  • the present disclosure provides methods for treating an adenocarcinoma with an EGFR activating mutation an EGFR copy number alteration (e.g., increased EGFR copy numbers), the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein in combination with a chemotherapy or a radiotherapy.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3
  • a subject having a cancer e.g., adenocarcinoma
  • an EGFR copy number alteration e.g., increased EGFR copy numbers
  • TKI tyrosine kinase inhibitors
  • a subject having a cancer e.g., adenocarcinoma
  • an EGFR copy number alteration e.g., increased EGFR copy numbers
  • TKI tyrosine kinase inhibitors
  • administering decreases ULBP2 expression level in a cancer with an EGFR activating mutation an EGFR copy number alteration (e.g., increased EGFR copy numbers) and the combination therapy with a chemotherapy, radio therapy, and/or immune checkpoint inhibitor improves the efficacy of the anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3).
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3 provided herein
  • an anti-ULBP2 antibody decreases ULBP2 expression level in a cancer with an EGFR activating mutation an EGFR copy number alteration (e.g., increased EGFR copy numbers) and the combination therapy with a chemotherapy, radio therapy, and/or immune checkpoint inhibitor improves the efficacy of the anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3).
  • the disclosure provides a method of treating a metastatic cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3) provided herein in combination with a chemotherapy, a radio therapy, and/or an immune checkpoint inhibitor.
  • an anti-ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3 provided herein in combination with a chemotherapy, a radio therapy, and/or an immune checkpoint inhibitor.
  • Metastatic cancers are caused by tumors that have spread from their original site (non-primary tumors) to other parts of the body. Non-primary tumors can metastasize via the bloodstream or lymphatic system. Metastatic cancers are typically more advanced and often harder to treat, as they can invade organs such as the lungs, liver, bones, or brain.
  • metastatic squamous cell carcinoma of the lung and cervical squamous cell carcinoma are especially prevalent.
  • Non-primary tumors are identified based on their cellular characteristics and location relative to the putative primary tumor. Methods to identify non- primary tumors include histopathology, immunohistochemistry, imaging e.g., CT, MRI, or PET scans), genomic profiling (e.g., DNA sequencing), and medical history. In some embodiments, metastatic cancer having non-primary tumors have a similar ULBP2 expression level as their primary tumor cells.
  • an anti- ULBP2 antibody e.g., a bispecific antibody that binds ULBP2 and CD3 provided herein
  • administration of an anti- ULBP2 antibody decreases ULBP2 expression level in a non-primary metastatic cancer and the combination therapy with a chemotherapy, radio therapy, and/or immune checkpoint inhibitor improves the efficacy of the anti-ULBP2 antibody (e.g., a bispecific antibody that binds ULBP2 and CD3).
  • the disclosure provides a method of treating a metastatic cancer in a subject, the method comprising administering to the subject an anti-ULBP2 antibody provided herein and a chemotherapy agent, a radiation therapy and/or an immune checkpoint inhibitor.
  • compositions suitable for administration can be incorporated into pharmaceutical compositions suitable for administration.
  • Such compositions typically comprise the antibody and a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the most recent edition of Remington’s Pharmaceutical Sciences, a standard reference text in the field, which is incorporated herein by reference.
  • Such carriers or diluents include, but are not limited to, water, saline, ringer’s solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used.
  • the use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.
  • a pharmaceutical composition of the disclosure is formulated to be compatible with its intended route of administration.
  • routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral e.g., inhalation), transdermal (z.e., topical), transmucosal, and rectal administration.
  • Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose.
  • the pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide.
  • the parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
  • compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
  • suitable carriers include physiological saline, bacteriostatic water, Cremophor ELTM (BASF, Parsippany, N J.) or phosphate buffered saline (PBS).
  • the composition must be sterile and should be fluid to the extent that easy syringeability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof.
  • the proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like.
  • Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above.
  • methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
  • Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition.
  • the tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or com starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
  • a binder such as microcrystalline cellulose, gum tragacanth or gelatin
  • an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or com starch
  • a lubricant such as magnesium stearate or Sterotes
  • a glidant such as colloidal silicon dioxide
  • the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.
  • a suitable propellant e.g., a gas such as carbon dioxide, or a nebulizer.
  • Systemic administration can also be by transmucosal or transdermal means.
  • penetrants appropriate to the barrier to be permeated are used in the formulation.
  • penetrants are generally known in the art, and include, for example, transmucosal administration, detergents, bile salts, and fusidic acid derivatives.
  • Transmucosal administration can be accomplished through the use of nasal sprays or suppositories.
  • the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art.
  • the compounds can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.
  • Dosage unit form refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.
  • the specification for the dosage unit forms of the disclosure are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.
  • compositions can be included in a container, pack, or dispenser together with instructions for administration.
  • Enzymatic reactions and purification techniques are performed according to manufacturer's specifications or as commonly accomplished in the art or as described herein.
  • the foregoing techniques and procedures are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification. See e.g., Sambrook et al. Molecular Cloning: A Laboratory Manual (2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989)).
  • the nomenclatures utilized in connection with, and the laboratory procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well-known and commonly used in the art. Standard techniques are used for chemical syntheses, chemical analyses, pharmaceutical preparation, formulation, and delivery, and treatment of patients.
  • ULBP2-positive cancer refers to a cancer comprising cells that express ULBP2 on their surface.
  • expression of ULBP2 on the cell surface is determined, for example, using antibodies to ULBP2 in a method such as immunohistochemistry, LACS, etc.
  • ULBP2 mRNA expression is considered to correlate to ULBP2 expression on the cell surface and can be determined by a method selected from in situ hybridization and RT-PCR (including quantitative RT-PCR).
  • ULBP2-positive cell refers to a cell that expresses ULBP2 on its surface.
  • cancer and “cancerous” refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth/proliferation.
  • cancer examples include, but are not limited to, carcinoma, lymphoma (e.g., Hodgkin's and non-Hodgkin's lymphoma), blastoma, sarcoma, and leukemia. More particular examples of such cancers include squamous cell cancer, small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastrointestinal cancer, pancreatic cancer, glioma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer (including triple negative (ER-/PR-/Her2-) breast cancer), colon cancer, colorectal cancer, endometrial or uterine carcinoma, salivary gland carcinoma, kidney cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, leukemia and other lymphoproliferative disorders, and various types of head and neck cancer
  • reference sample denotes a sample with at least one known characteristic that can be used as a comparison to a sample with at least one unknown characteristic.
  • a reference sample can be used as a positive or negative indicator.
  • a reference sample can be used to establish a level of protein and/or mRNA that is present in, for example, healthy tissue, in contrast to a level of protein and/or mRNA present in the sample with unknown characteristics.
  • the reference sample comes from the same subject, but is from a different part of the subject than that being tested.
  • the reference sample is from a tissue area surrounding or adjacent to the cancer.
  • the reference sample is not from the subject being tested, but is a sample from a subject known to have, or not to have, a disorder in question (for example, a particular cancer or ULBP2 related disorder).
  • the reference sample is from the same subject, but from a point in time before the subject developed cancer.
  • the reference sample is from a benign cancer sample (for example, benign breast cancer sample), from the same or a different subject.
  • a negative reference sample is used for comparison, the level of expression or amount of the molecule in question in the negative reference sample will indicate a level at which one of skill in the art will appreciate, given the present disclosure, that there is no and/or a low level of the molecule.
  • the level of expression or amount of the molecule in question in the positive reference sample will indicate a level at which one of skill in the art will appreciate, given the present disclosure, that there is a level of the molecule.
  • the term “disease” or “disorder” as used herein refers to a condition where treatment is needed and/or desired.
  • the term “tumor cell”, “cancer cell”, “cancer”, “tumor”, and/or “neoplasm”, unless otherwise designated, are used herein interchangeably and refer to a cell (or cells) exhibiting an uncontrolled growth and/or abnormal increased cell survival and/or inhibition of apoptosis which interferes with the normal functioning of bodily organs and systems. Included in this definition are benign and malignant cancers, polyps, hyperplasia, as well as dormant tumors or micrometastases.
  • cancer and “tumor” encompass solid and hematological/lymphatic cancers and also encompass malignant, pre-malignant, and benign growth, such as dysplasia. Also, included in this definition are cells having abnormal proliferation that is not impeded (e g immune evasion and immune escape mechanisms) by the immune system (e.g. virus infected cells).
  • Exemplary tumor cells include, but are not limited to: basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and central nervous system cancer; breast cancer; cancer of the peritoneum; cervical cancer; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer (including gastrointestinal cancer); glioblastoma; hepatic carcinoma; hepatoma; intra-epithelial neoplasm; kidney or renal cancer; larynx cancer; leukemia; liver cancer; lung cancer e.g., small-cell lung cancer, non- small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung); melanoma; myeloma; neuroblastoma; oral cavity cancer (lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer;
  • the term cancer can encompass: a lung cancer, a breast cancer, a head and neck cancer, an ovarian cancer, and/or an endometrial cancer.
  • a lung cancer a breast cancer, a head and neck cancer, an ovarian cancer, and/or an endometrial cancer.
  • the difference between a “cancer” and a “cancer cell” can be denoted by the use of the explicit use of the phrase “cancer cell”; however, the term “cancer” will encompass concepts such as the subject having cancer, and multicellular tumors, as well as single cancer cells.
  • an “increase or decrease” refers to a statistically significant increase or decrease respectively.
  • “modulating” can also involve effecting a change (which can either be an increase or a decrease) in affinity, avidity, specificity and/or selectivity of a target or antigen, for one or more of its ligands, binding partners, partners for association into a homomultimeric or heteromultimeric form, or substrates; effecting a change (which can either be an increase or a decrease) in the sensitivity of the target or antigen for one or more conditions in the medium or surroundings in which the target or antigen is present (such as pH, ion strength, the presence of co-factors, etc.), compared to the same conditions but without the presence of an antibody, bispecific or multispecific polypeptide agent.
  • This can be determined in any suitable manner and/or using any suitable assay known per se or described herein, depending on the target involved.
  • an immune response is meant to encompass cellular and/or humoral immune responses that are sufficient to inhibit or prevent onset or ameliorate the symptoms of disease (for example, cancer or cancer metastasis).
  • An immune response can encompass aspects of both the innate and adaptive immune systems.
  • treatment is an approach for obtaining beneficial or desired clinical results.
  • Treatment covers any administration or application of a therapeutic for disease in a mammal, including a human.
  • beneficial or desired clinical results include, but are not limited to, any one or more of: alleviation of one or more symptoms, diminishment of extent of disease, preventing or delaying spread (for example, metastasis, for example metastasis to the lung or to the lymph node) of disease, preventing or delaying recurrence of disease, delay or slowing of disease progression, amelioration of the disease state, inhibiting the disease or progression of the disease, inhibiting or slowing the disease or its progression, arresting its development, and remission (whether partial or total).
  • “Ameliorating” means a lessening or improvement of one or more symptoms as compared to not administering an anti-ULBP2 antibody. “Ameliorating” also includes shortening or reduction in duration of a symptom.
  • biological sample means a quantity of a substance from a living thing or formerly living thing.
  • substances include, but are not limited to, blood, (for example, whole blood), plasma, serum, urine, amniotic fluid, synovial fluid, endothelial cells, leukocytes, monocytes, other cells, organs, tissues, bone marrow, lymph nodes and spleen.
  • control refers to a composition known to not contain an analyte (“negative control”) or to contain analyte (“positive control”).
  • a positive control can comprise a known concentration of analyte.
  • Control “positive control,” and “calibrator” may be used interchangeably herein to refer to a composition comprising a known concentration of analyte.
  • a “positive control” can be used to establish assay performance characteristics and is a useful indicator of the integrity of reagents (for example, analytes).
  • Predetermined cutoff and “predetermined level” refer generally to an assay cutoff value that is used to assess diagnostic/prognostic/therapeutic efficacy results by comparing the assay results against the predetermined cutoff/level, where the predetermined cutoff/level already has been linked or associated with various clinical parameters (for example, severity of disease, progression/nonprogression/improvement, etc.). While the present disclosure may provide exemplary predetermined levels, it is well-known that cutoff values may vary depending on the nature of the immunoassay (for example, antibodies employed, etc.).
  • inhibitortion refers to a decrease or cessation of any phenotypic characteristic or to the decrease or cessation in the incidence, degree, or likelihood of that characteristic.
  • To “reduce” or “inhibit” is to decrease, reduce or arrest an activity, function, and/or amount as compared to a reference.
  • by “reduce” or “inhibit” is meant the ability to cause an overall decrease of 20% or greater.
  • by “reduce” or “inhibit” is meant the ability to cause an overall decrease of 50% or greater.
  • by “reduce” or “inhibit” is meant the ability to cause an overall decrease of 75%, 85%, 90%, 95%, or greater.
  • the amount noted above is inhibited or decreased over a period of time, relative to a control dose (such as a placebo) over the same period of time.
  • “delaying development of a disease” means to defer, hinder, slow, retard, stabilize, suppress and/or postpone development of the disease (such as cancer). This delay can be of varying lengths of time, depending on the history of the disease and/or individual being treated. As is evident to one skilled in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the individual does not develop the disease. For example, a late stage cancer, such as development of metastasis, may be delayed.
  • Preventing includes providing prophylaxis with respect to the occurrence or recurrence of a disease in a subject that may be predisposed to the disease but has not yet been diagnosed with the disease. Unless otherwise specified, the terms “reduce”, “inhibit”, or “prevent” do not denote or require complete prevention over all time.
  • to “suppress” a function or activity is to reduce the function or activity when compared to otherwise same conditions except for a condition or parameter of interest, or alternatively, as compared to another condition.
  • an antibody which suppresses tumor growth reduces the rate of growth of the tumor compared to the rate of growth of the tumor in the absence of the antibody.
  • antibody binding region refers to a region of the antigen, which comprises the epitope to which the antibody binds.
  • An antibody binding region may be determined by epitope binning using biolayer interferometry, by alanine scan, or by domain shuffle assays (using antigen constructs in which regions of the antigen are exchanged with that of another species and determining whether the antibody still binds to the antigen or not).
  • the amino acids within the antibody binding region that are involved in the interaction with the antibody may be determined by hydrogen/deuterium exchange mass spectrometry and/or by crystallography of the antibody bound to its antigen.
  • the term “antibody” refers to immunoglobulin molecules and immunologically active portions of immunoglobulin (Ig) molecules, i.e., molecules that contain an antigen binding site that specifically binds (immunoreacts with) an antigen.
  • immunoglobulin immunoglobulin molecules
  • immunologically active portions of immunoglobulin (Ig) molecules i.e., molecules that contain an antigen binding site that specifically binds (immunoreacts with) an antigen.
  • Ig immunoglobulin
  • Antibodies include, but are not limited to, polyclonal, monoclonal, chimeric, dAb (domain antibody), single chain, Fab, Fab’ and F( a b')2 fragments, scFvs, and a F a b expression library.
  • the basic antibody structural unit is known to comprise a tetramer.
  • Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one “light” (about 25 kDa) and one “heavy” chain (about 50-70 kDa).
  • the amino-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition.
  • the carboxy-terminal portion of each chain defines a constant region primarily responsible for effector function.
  • antibody molecules obtained from humans relate to any of the classes IgG, IgM, IgA, IgE and IgD, which differ from one another by the nature of the heavy chain present in the molecule. Certain classes have subclasses as well, such as IgGi, IgG2, IgG4 and others.
  • the light chain may be a kappa chain or a lambda chain.
  • the complementarity determining regions (CDRs) of the monoclonal antibody are identical in all the molecules of the population.
  • MAbs contain an antigen binding site capable of immunoreacting with a particular epitope of the antigen characterized by a unique binding affinity for it.
  • antigen binding region or “antigen-binding site” or “binding portion” refers to the part of the immunoglobulin molecule that participates in antigen binding.
  • the antigen binding site is formed by amino acid residues of the N-terminal variable (“V”) regions of the heavy (“H”) and light (“L”) chains.
  • V N-terminal variable
  • H heavy
  • L light
  • hypervariable regions Three highly divergent stretches within the V regions of the heavy and light chains, referred to as “hypervariable regions,” are interposed between more conserved flanking stretches known as “framework regions,” or “FRs”.
  • FR refers to amino acid sequences which are naturally found between, and adjacent to, hypervariable regions in immunoglobulins.
  • the three hypervariable regions of a light chain and the three hypervariable regions of a heavy chain are disposed relative to each other in three dimensional space to form an antigen-binding surface.
  • the antigen-binding surface is complementary to the three-dimensional surface of a bound antigen, and the three hypervariable regions of each of the heavy and light chains are referred to as “complementarity-determining regions,” or “CDRs.”
  • CDRs complementarity-determining regions
  • the Kabat numbering system See Kabat, E.A., et al., Sequences of Protein of immunological interest, Fifth Edition, US Department of Health and Human Services, US Government Printing Office (1991)
  • the IMGT numbering system See IMGT®, the international ImMunoGeneTics information system®. Available online: http://www.imgt.org/).
  • the IMGT numbering system is routinely used and accepted as a reliable and accurate system in the art to determine amino acid positions in coding sequences, alignment of alleles, and to easily compare sequences in immunoglobulin (IG) and T-cell receptor (TR) from all vertebrate species.
  • IG immunoglobulin
  • TR T-cell receptor
  • IMGT-ONTOLOGY the first, and so far unique, ontology for immunogenetics and immunoinformatics (See Lefranc. M.P. et al., Biomolecules, 2014 Dec; 4(4), 1102-1139).
  • IMGT tools and databases run against IMGT reference directories built from a large repository of sequences.
  • the IG V-DOMAIN and IG C-DOMAIN are delimited taking into account the exon delimitation, whenever appropriate.
  • the IMGT exon numbering system can be and “is used” by those skilled in the art reliably to determine amino acid positions in coding sequences and for alignment of alleles. Additionally, correspondences between the IMGT unique numbering with other numberings (i.e., Kabat) are available in the IMGT Scientific chart (See Lefranc. M.P. et al., Biomolecules, 2014 Dec; 4(4), 1102-1139).
  • hypervariable region refers to the amino acid residues of an antibody that are typically responsible for antigen-binding.
  • the hypervariable region generally comprises amino acid residues from a "complementarity determining region” or "CDR" (e.g., around about residues 24-34 (LI), 50-56 (L2) and 89-97 (L3) in the VL, and around about 31-35 (HI), 50-65 (H2) and 95-102 (H3) in the Vn when numbered in accordance with the Kabat numbering system; Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md.
  • CDR complementarity determining region
  • residues from a "hypervariable loop” e.g., residues 24- 34 (LI), 50-56 (L2) and 89-97 (L3) in the VL, and 26-32 (HI), 52-56 (H2) and 95-101 (H3) in the VH when numbered in accordance with the Chothia numbering system; Chothia and Lesk, J. Mol. Biol.
  • residues from a "hypervariable loop" VCDR e.g., residues 27-38 (LI), 56-65 (L2) and 105-120 (L3) in the VL, and 27-38 (HI), 56-65 (H2) and 105-120 (H3) in the Vn when numbered in accordance with the IM GT numbering system; Lefranc, M.P. et al. Nucl. Acids Res. 27:209-212 (1999), Ruiz, M. e al. Nucl. Acids Res. 28:219-221 (2000)).
  • a "hypervariable loop" VCDR e.g., residues 27-38 (LI), 56-65 (L2) and 105-120 (L3) in the VL, and 27-38 (HI), 56-65 (H2) and 105-120 (H3) in the Vn when numbered in accordance with the IM GT numbering system; Lefranc, M.P. et al. Nucl. Acids Res. 27
  • the antibody has symmetrical insertions at one or more of the following points 28, 36 (LI), 63, 74-75 (L2) and 123 (L3) in the VL, and 28, 36 (HI), 63, 74-75 (H2) and 123 (H3) in the VH when numbered in accordance with AHo; Honneger, A. and Plunkthun, A. J. Mol. Biol. 309:657-670 (2001)).
  • epitopic determinants include any protein determinant capable of specific binding to an immunoglobulin, an scEv, or a T-cell receptor.
  • epitopic determinants usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics.
  • antibodies may be raised against N-terminal or C-terminal peptides of a polypeptide.
  • An antibody is the to specifically bind an antigen when the dissociation constant is ⁇ 1 pM; e.g., ⁇ 100 nM, preferably ⁇ 10 nM and more preferably ⁇ 1 nM.
  • immunological binding refers to the non-covalent interactions of the type which occur between an immunoglobulin molecule and an antigen for which the immunoglobulin is specific.
  • the strength, or affinity of immunological binding interactions can be expressed in terms of the dissociation constant (Ka) of the interaction, wherein a smaller Ka represents a greater affinity.
  • Immunological binding properties of selected polypeptides can be quantified using methods well known in the art. One such method entails measuring the rates of antigen-binding site/antigen complex formation and dissociation, wherein those rates depend on the concentrations of the complex partners, the affinity of the interaction, and geometric parameters that equally influence the rate in both directions.
  • both the “on rate constant” (K on ) and the “off rate constant” (K o ff) can be determined by calculation of the concentrations and the actual rates of association and dissociation. (See Nature 361:186-87 (1993)).
  • the ratio of K o ff /Kon enables the cancellation of all parameters not related to affinity, and is equal to the dissociation constant Ka. (See, generally, Davies et al. (1990) Annual Rev Biochem 59:439-473).
  • An antibody of the present disclosure is the to specifically bind to its target, when the equilibrium binding constant (Ka) is ⁇ 1 p.M, e.g., ⁇ 100 nM, preferably ⁇ 10 nM, and more preferably ⁇ 1 nM, as measured by assays such as radioligand binding assays or similar assays known to those skilled in the art.
  • Ka equilibrium binding constant
  • isolated polynucleotide shall mean a polynucleotide of genomic, cDNA, or synthetic origin or some combination thereof, which by virtue of its origin the “isolated polynucleotide” (1) is not associated with all or a portion of a polynucleotide in which the “isolated polynucleotide” is found in nature, (2) is operably linked to a polynucleotide which it is not linked to in nature, or (3) does not occur in nature as part of a larger sequence.
  • Polynucleotides in accordance with the disclosure include the nucleic acid molecules encoding the heavy chain immunoglobulin molecules, and nucleic acid molecules encoding the light chain immunoglobulin molecules described herein.
  • isolated protein means a protein of cDNA, recombinant RNA, or synthetic origin or some combination thereof, which by virtue of its origin, or source of derivation, the “isolated protein” (1) is not associated with proteins found in nature, (2) is free of other proteins from the same source, e.g., free of marine proteins, (3) is expressed by a cell from a different species, or (4) does not occur in nature.
  • polypeptide is used herein as a generic term to refer to native protein, fragments, or analogs of a polypeptide sequence. Hence, native protein fragments, and analogs are species of the polypeptide genus. Polypeptides in accordance with the disclosure comprise the heavy chain immunoglobulin molecules, and the light chain immunoglobulin molecules described herein, as well as antibody molecules formed by combinations comprising the heavy chain immunoglobulin molecules with light chain immunoglobulin molecules, such as kappa light chain immunoglobulin molecules, and vice versa, as well as fragments and analogs thereof.
  • the term “naturally-occurring” as used herein as applied to an object refers to the fact that an object can be found in nature. For example, a polypeptide or polynucleotide sequence that is present in an organism (including viruses) that can be isolated from a source in nature and which has not been intentionally modified by man in the laboratory or otherwise is naturally-occurring.
  • operably linked refers to positions of components so described are in a relationship permitting them to function in their intended manner.
  • a control sequence “operably linked” to a coding sequence is ligated in such a way that expression of the coding sequence is achieved under conditions compatible with the control sequences.
  • control sequence refers to polynucleotide sequences which are necessary to affect the expression and processing of coding sequences to which they are ligated. The nature of such control sequences differs depending upon the host organism in prokaryotes, such control sequences generally include promoter, ribosomal binding site, and transcription termination sequence in eukaryotes, generally, such control sequences include promoters and transcription termination sequence.
  • control sequences is intended to include, at a minimum, all components whose presence is essential for expression and processing, and can also include additional components whose presence is advantageous, for example, leader sequences and fusion partner sequences.
  • polynucleotide as referred to herein means a polymeric boron of nucleotides of at least 10 bases in length, either ribonucleotides or deoxynucleotides or a modified form of either type of nucleotide.
  • the term includes single and double stranded forms of DNA.
  • Examples of unconventional amino acids include: 4 hydroxyproline, y-carboxyglutamate, a-N,N,N-trimethyllysine, a -N- acetyllysine, O-phosphoserine, N- acetylserine, N-formylmethionine, 3-methylhistidine, 5- hydroxylysine, o-N-mcthylargininc, and other similar amino acids and imino acids (e.g., 4- hydroxyproline).
  • the left-hand direction is the amino terminal direction and the right-hand direction is the carboxy-terminal direction, in accordance with standard usage and convention.
  • the term “substantial identity” means that two peptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, share at least 80 percent sequence identity, preferably at least 90 percent sequence identity, more preferably at least 95 percent sequence identity, and most preferably at least 99 percent sequence identity.
  • residue positions which are not identical differ by conservative amino acid substitutions.
  • Conservative amino acid substitutions refer to the interchangeability of residues having similar side chains.
  • a group of amino acids having aliphatic side chains is glycine, alanine, valine, leucine, and isoleucine; a group of amino acids having aliphatic - hydroxyl side chains is serine and threonine; a group of amino acids having amide- containing side chains is asparagine and glutamine; a group of amino acids having aromatic side chains is phenylalanine, tyrosine, and tryptophan; a group of amino acids having basic side chains is lysine, arginine, and histidine; and a group of amino acids having sulfur- containing side chains is cysteine and methionine.
  • Preferred conservative amino acids substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine valine, glutamic- aspartic, and asparagine-glutamine.
  • amino acid sequences of antibodies or immunoglobulin molecules are contemplated as being encompassed by the present disclosure, providing that the variations in the amino acid sequence maintain at least 75%, more preferably at least 80%, 90%, 95%, and most preferably 99%.
  • conservative amino acid replacements are contemplated. Conservative replacements are those that take place within a family of amino acids that are related in their side chains.
  • amino acids are generally divided into families: (1) acidic amino acids are aspartate, glutamate; (2) basic amino acids are lysine, arginine, histidine; (3) non-polar amino acids are alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan, and (4) uncharged polar amino acids are glycine, asparagine, glutamine, cysteine, serine, threonine, tyrosine.
  • the hydrophilic amino acids include arginine, asparagine, aspartate, glutamine, glutamate, histidine, lysine, serine, and threonine.
  • the hydrophobic amino acids include alanine, cysteine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, tyrosine and valine.
  • Other families of amino acids include (i) serine and threonine, which are the aliphatic -hydroxy family; (ii) asparagine and glutamine, which are the amide containing family; (iii) alanine, valine, leucine and isoleucine, which are the aliphatic family; and (iv) phenylalanine, tryptophan, and tyrosine, which are the aromatic family.
  • Structural and functional domains can be identified by comparison of the nucleotide and/or amino acid sequence data to public or proprietary sequence databases.
  • computerized comparison methods are used to identify sequence motifs or predicted protein conformation domains that occur in other proteins of known structure and/or function. Methods to identify protein sequences that fold into a known three-dimensional structure are known. Bowie et al. Science 253:164 (1991).
  • sequence motifs and structural conformations that may be used to define structural and functional domains in accordance with the disclosure.
  • Preferred amino acid substitutions are those which: (1) reduce susceptibility to proteolysis, (2) reduce susceptibility to oxidation, (3) alter binding affinity for forming protein complexes, (4) alter binding affinities, and (4) confer or modify other physicochemical or functional properties of such analogs.
  • Analogs can include various muteins of a sequence other than the naturally-occurring peptide sequence. For example, single or multiple amino acid substitutions (preferably conservative amino acid substitutions) may be made in the naturally- occurring sequence (preferably in the portion of the polypeptide outside the domain(s) forming intermolecular contacts.
  • a conservative amino acid substitution should not substantially change the structural characteristics of the parent sequence (e.g., a replacement amino acid should not tend to break a helix that occurs in the parent sequence, or disrupt other types of secondary structure that characterizes the parent sequence).
  • Examples of art-recognized polypeptide secondary and tertiary structures are described in Proteins, Structures and Molecular Principles (Creighton, Ed., W. H. Freeman and Company, New York (1984)); Introduction to Protein Structure (C. Branden and J. Tooze, eds., Garland Publishing, New York, N.Y. (1991)); and Thornton et at. Nature 354:105 (1991).
  • label refers to incorporation of a detectable marker, e.g., by incorporation of a radiolabeled amino acid or attachment to a polypeptide of biotinyl moieties that can be detected by marked avidin (e.g., streptavidin containing a fluorescent marker or enzymatic activity that can be detected by optical or calorimetric methods). In certain situations, the label or marker can also be therapeutic. Various methods of labeling polypeptides and glycoproteins are known in the art and may be used.
  • labels for polypeptides include, but are not limited to, the following: radioisotopes or radionuclides (e.g., 3 H, 14 C, 15 N, 35 S, 90 Y, "Tc, n i In, 125 I, 131 I), fluorescent labels (e.g., FITC, rhodamine, lanthanide phosphors), enzymatic labels (e.g., horseradish peroxidase, p- galactosidase, luciferase, alkaline phosphatase), chemiluminescent, biotinyl groups, predetermined polypeptide epitopes recognized by a secondary reporter (e.g., leucine zipper pair sequences, binding sites for secondary antibodies, metal binding domains, epitope tags).
  • radioisotopes or radionuclides e.g., 3 H, 14 C, 15 N, 35 S, 90 Y, "Tc, n i In, 125 I,
  • labels are attached by spacer arms of various lengths to reduce potential steric hindrance.
  • pharmaceutical agent or drug refers to a chemical compound or composition capable of inducing a desired therapeutic effect when properly administered to a patient.
  • substantially pure means an object species is the predominant species present (i.e., on a molar basis it is more abundant than any other individual species in the composition), and preferably a substantially purified fraction is a composition wherein the object species comprises at least about 50 percent (on a molar basis) of all macromolecular species present.
  • a substantially pure composition will comprise more than about 80 percent of all macromolecular species present in the composition, more preferably more than about 85%, 90%, 95%, and 99%.
  • the object species is purified to essential homogeneity (contaminant species cannot be detected in the composition by conventional detection methods) wherein the composition consists essentially of a single macromolecular species.
  • patient includes human and veterinary subjects.
  • Example 1 Enrichment of ULBP2 in the basal/squamous lineage that is inherent to specific subtypes of cancer
  • A549, 5637, CORL105, FADU, NCIH292 cell lines were obtained from ATCC and Sigma. They were engineered with GFP-Luc via lentivirus (BPS) for ease of tracking in killing assays.
  • Cells were lysed in RIPA buffer with Halt protease and phosphatase inhibitor cocktail (Pierce), and then quantified for protein content using a BCA assay (Pierce). Lysates were normalized, denatured and reduced in 4X SDS buffer (Pierce) and run on 4-12% NuPAGE Bis-Tris SDS-PAGE. Gels were transferred onto PVDF membranes (iBlot2) and blocked in LiCOR blocking buffer.
  • Example 2 Basal/squamous Indication or subtypes showing enrichment of ULBP2 have higher immune infiltration
  • the gene SERP1NB3 that has been associated with squamous indications at the soluble level is also clearly higher in the basal/squamous subtype (FIG. 4D). Furthermore, the basal/squamous subtype is associated with immune infiltration as can be seen by enrichment of PD-L1 (FIG. 5A), PD1 (FIG. 5B), CD8A (FIG. 5C), and Granzyme B (FIG. 5D) over the luminal subtypes. This means that not only was ULBP2 enriched in the basal/squamous subtype of bladder cancer, but those patients also had increased immune infiltration in their tumor, making it an ideal tumor microenvironment for a T-cell engager.
  • NSCLC For lung cancer, histological assessment defines NSCLC into either adenocarcinoma or squamous cell carcinoma. Gene expression normalized reads were aggregated from both the TCGA adenocarcinoma and squamous cell carcinoma into a larger NSCLC subset. From this data, when broken up by indication, ULBP2 (FIG. 6A) was much higher expressed in the squamous cell indication. This is less true for TROP2 (FIG. 6B) and NECTIN4 (FIG. 6D), while HER2 (FIG. 6C) was much more enriched in lung adenocarcinoma, consistent with it being used as a luminal marker.
  • FIG. 7A luminal genes FOXA1 (FIG. 7A) and PPARG (FIG. 7B) and the squamous gene KRT5 (FIG. 7C).
  • SERPINB3 also clearly showed an enrichment in lung squamous cell and not lung adenocarcinoma (FIG. 7D). While lung cancer did not show as much infiltration in the basal/squamous indication, there was some trend towards more immune infiltration in that indication (PD-L1: FIG. 8A; Granzyme B: FIG. 8D) and even those that were slightly higher in adenocarcinoma (PD1: FIG. 8B; CD8A: FIG.
  • Example 3 ULBP2 targeted therapies are improved with combinations with PD1/PDL1 blockade.
  • PBMC-mediated cytotoxicity assays [0511] PBMCs were isolated from healthy donor leukopaks following standard procedure and frozen. Human tumor cells were seeded at 10,000 cells per well in 96-well tissue culture plates (Perkin Elmer) and incubated for 24 hours at 37 C with 5 % CO2. On the following day, PBMCs were thawed using benzonase and two spins with complete media. PBMCs (150,000) were added to tumor cells in the presence of titrated EIP0820 or EIP0607 control antibodies and either pembrolizumab (IgG4 antibody against PD1 used extensively in the clinic) or the IgG4 control at 2 ug/mL, and incubated for up to 5 days.
  • EIP0820 or EIP0607 control antibodies either pembrolizumab (IgG4 antibody against PD1 used extensively in the clinic) or the IgG4 control at 2 ug/mL
  • Example 4 ULBP2 levels are increased with specific chemotherapies that impart DNA damage to the cell; thereby, ULBP2 targeted therapies are improved in combination with specific chemotherapy agents.
  • Cell lines (A549, 5637) were plated at 10,000 per well in a 96-well plate. 24 h after seeding, chemotherapies at a range of concentrations were added to the cells. After 72 h, the cells were read out using CellTiter Gio (Promega) to determine EC50 of cell arrest/killing. At those EC50s for each chemotherapy and each cell line, surface levels of ULBP2, TROP2, HER2, CD58, PDL1, and B7H4 were determined using analytical flow cytometry (BD Symphony A5) and an 8-color panel. Appropriate isotype controls for each antibody were also utilized to subtract background fluorescence levels. Samples were analyzed on FlowJo and graphed in GraphPad Prism.
  • ULBP2 was the most differentially upregulated in two different cell lines (FIG. 10A-10B).
  • chemotherapies that induced a DNA damage response such as the topoisomerase inhibitor irinotecan, anthracycline doxorubicin, DNA break inducing antibiotic bleomycin, pyrimidine antagonist gemcitabine (“gem”), and platinum alkylating agent cisplatin (“cis”) increased ULBP2 levels significantly as compared to the microtubule-targeting taxane docetaxel and vinca alkaloid vinblastine.
  • chemotherapies increased PDL1 non- specifically in both cell lines and a few other targets were specifically induced by certain chemotherapies but only in one cell line. Therefore, patient treatment of specific DNA-damaging chemotherapies can increase efficacy of ULBP2-targeting biologies either in cotreatment or subsequent treatment.
  • EIP0820 Concurrent or sequential administration of EIP0820 with chemotherapy and checkpoint inhibitors can increase UEBP2 expression and thereby EIP0820 efficacy.
  • Another standard of care, used in both lung and bladder cancers, that can be combined with EIP0820 is adaptive radiotherapy.
  • Previous publications (Gasser et al. Nature Fetters 2005 doi:10.1038/nature03884) confirm increased NKG2D ligands, including UEBP2, after in vitro exposure to ionizing radiation. Therefore, the data indicate that combinations subsequently or concurrently with multiple therapies are possible and warranted for EIP0820.
  • Example 5 Expression of ULBP2 is associated with squamous subtypes within indication and between indications.
  • esophageal cancer could also be histologically divided into adenocarcinoma and squamous cell carcinoma. Evaluating ULBP2 levels in patient tumor samples showed that expression is well correlated with squamous subtype in this indication (FIG. 13).
  • any squamous indications (such as thymic squamous cell carcinoma, anal squamous cell carcinoma, vulvar carcinoma), including ones of rare nature that may not be part of TCGA, likely also have high UEBP2 expression and can therefore be easily targeted by EIP0820.
  • ULBP2 is strongly associated with basal/squamous molecular subtype and CD8 T cell infiltration.
  • Example 6 Expression of ULBP2 in tumors does not change in metastatic samples.
  • Example 7 Tumor ULBP2 expression is higher in HPV negative patients in head and neck cancers.
  • Example 8 Patients with higher stage/grade have higher tumor ULBP2.
  • Example 9 Standard of Care (SOC) of cancer do not decrease tumor ULBP2 expression levels in patients.
  • FIGs. 18A-18B Tumors from patients treated with checkpoint inhibitors or chemotherapy were assessed for whether ULBP2 levels change after treatment.
  • treatment with the anti-PDl checkpoint inhibitor nivolumab did not change average ULBP2 levels (FIGs. 18A-18B).
  • FIG. 18C For lung cancer datasets, either paired (FIG. 18C) or unpaired (FIG. 18D), there was no change in ULBP2 level in the tumor with various PD1 and PDL1 checkpoint inhibitors.
  • FIG. 18C paired
  • FIG. 18D unpaired
  • there was no change in ULBP2 level in the tumor with various PD1 and PDL1 checkpoint inhibitors For lung cancer dataset GSE248378, squamous and adeno pathology subtypes were divided, and the effect of the anti-PDLl checkpoint inhibitor durvalumab was assessed. A significant change in either subtype was not observed (FIGs.
  • Example 10 Patients with EGER mutations and patients that ultimately receive TKIs have higher ULBP2 levels.
  • Lung adenocarcinoma patients that have EGFR mutations typically receive tyrosine kinase inhibitors (TKIs). While ULBP2 expression was not observed to be significantly higher in patient tumors with higher EGFR expression (FIG. 19A), it was significantly higher in patient tumors with EGFR mutations, as seen in the TCGA dataset (FIG. 19B). It was also higher in patients with EGFR copy number alterations (FIG. 19C). Most of the patients with mutations have activating EGFR mutations and go on to receive TKIs.
  • TKIs tyrosine kinase inhibitors
  • Example 11 Squamous subtypes in bladder cancer have more CD3+ T cells
  • TMA tumor microarray
  • TAA tumor microarray of bladder cancer
  • Biocare Medical EP41 an anti-CD3 antibody
  • the staining was quantified to determine a CD3+ cell density score for each core.
  • squamous cell indications not only had higher ULBP2/5/6 expression, but also had higher CD3+ T cell infiltration (as described in Example 3), which are ideal conditions for treatment with for the T cell engager EIP0820.
  • Example 12 Rare squamous cancers also have high ULBP2 expression.
  • Vulvar cancer samples had higher ULBP2 expression than normal vulvar tissue and higher UBP2 expression in the tumor than either cervical or endometrial carcinomas (FIG. 21B). Furthermore, the association between ULBP2 expression and HPV negative status observed in head and neck cancers also held true in vulvar carcinoma (FIG. 21C). The extension of ULBP2 expression to these rarer squamous cancers indicates that EIP0820 can be efficacious in any squamous indication, including not only anal squamous cell carcinoma and vulvar cancer, but also penile cancer, thymic squamous cell carcinoma, skin cutaneous squamous cell carcinoma, and others.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Epidemiology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente divulgation concerne des anticorps anti-ULBP2 ou des fragments de liaison à l'antigène de ceux-ci, notamment des bispécifiques. Les anticorps sont administrés en combinaison avec un agent de chimiothérapie, une radiothérapie et/ou un inhibiteur de point de contrôle immunitaire pour le traitement du cancer épidermoïde basal. La présente divulgation concerne également des méthodes et des kits pour détecter une prédisposition à des troubles liés à ULBP2, déterminer le risque de développer des troubles liés à ULBP2 et guider une thérapie contre des troubles liés à ULBP2.
PCT/US2024/051568 2023-10-16 2024-10-16 Polythérapies avec des anticorps ciblant ulbp2 pour traiter des cancers Pending WO2025085512A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202363590755P 2023-10-16 2023-10-16
US63/590,755 2023-10-16

Publications (2)

Publication Number Publication Date
WO2025085512A1 true WO2025085512A1 (fr) 2025-04-24
WO2025085512A9 WO2025085512A9 (fr) 2025-06-05

Family

ID=93432419

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2024/051568 Pending WO2025085512A1 (fr) 2023-10-16 2024-10-16 Polythérapies avec des anticorps ciblant ulbp2 pour traiter des cancers

Country Status (1)

Country Link
WO (1) WO2025085512A1 (fr)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3773919A (en) 1969-10-23 1973-11-20 Du Pont Polylactide-drug mixtures
US4522811A (en) 1982-07-08 1985-06-11 Syntex (U.S.A.) Inc. Serial injection of muramyldipeptides and liposomes enhances the anti-infective activity of muramyldipeptides
WO2023178357A1 (fr) * 2022-03-18 2023-09-21 Evolveimmune Therapeutics, Inc. Molécules de fusion d'anticorps bispécifiques et leurs procédés d'utilisation

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3773919A (en) 1969-10-23 1973-11-20 Du Pont Polylactide-drug mixtures
US4522811A (en) 1982-07-08 1985-06-11 Syntex (U.S.A.) Inc. Serial injection of muramyldipeptides and liposomes enhances the anti-infective activity of muramyldipeptides
WO2023178357A1 (fr) * 2022-03-18 2023-09-21 Evolveimmune Therapeutics, Inc. Molécules de fusion d'anticorps bispécifiques et leurs procédés d'utilisation

Non-Patent Citations (44)

* Cited by examiner, † Cited by third party
Title
"Drug Absorption Enhancement: Concepts, Possibilities, Limitations, And Trends", 1994, HARWOOD ACADEMIC PUBLISHERS
"ELISA: Theory and Practice: Methods in Molecular Biology", vol. 42, 1995, MACK PUB. CO.
"Peptide And Protein Drug Delivery", vol. 4, 1991, M. DEKKER, article "Advances In Parenteral Sciences"
"Proteins, Structures and Molecular Principles", 1984, W. H. FREEMAN AND COMPANY
"Remington's Pharmaceutical Sciences", 1975, MACK PUBLISHING COMPANY
BAKER, D. J. ET AL.: "Clearance of pl6Ink4a-positive senescent cells delays ageing-associated disorders", NATURE, vol. 479, 2011, pages 232 - 236, XP055074325, DOI: 10.1038/nature10600
BAKER, D. J. ET AL.: "Naturally occurring pl6Ink4a-positive cells shorten healthy lifespan", NATURE, vol. 530, 2016, pages 184 - 189, XP093027320, DOI: 10.1038/nature16932
BALDRICK P.: "Pharmaceutical excipient development: the need for preclinical guidance", REGUL. TOXICOL PHARMACOL., vol. 32, no. 2, 2000, pages 210 - 8
BOWIE ET AL., SCIENCE, vol. 253, 1991, pages 164
CHARMAN WN: "Lipids, lipophilic drugs, and oral drug delivery-some emerging concepts", J PHARM SCI., vol. 89, no. 8, 2000, pages 967 - 78, XP008099512
CHILDS, B. G. ET AL.: "Senescent cells: an emerging target for diseases of ageing", NAT. REV. DRUG DISCOV., vol. 16, 2017, pages 718 - 735, XP055979258, DOI: 10.1038/nrd.2017.116
CHOTHIALESK, J. MOL. BIOL., vol. 196, 1987, pages 901 - 917
CRAGG, M. S. ET AL., BLOOD, vol. 101, 2003, pages 1045 - 1052
CRAGG, M. S. ET AL., BLOOD, vol. 103, 2004, pages 2738 - 2743
CUNNINGHAMWELLS, SCIENCE, vol. 244, 1989, pages 1081 - 1085
DAVIES ET AL., ANNUAL REV BIOCHEM, vol. 59, 1990, pages 439 - 473
DEMARIA, M. ET AL.: "Cellular Senescence Promotes Adverse Effects of Chemotherapy and Cancer Relapse", CANCER DISCOV., vol. 7, 2017, pages 165 - 176
E. DIAMANDIST. CHRISTOPOULUS: "Immunoassay", 1996, ACADEMIC PRESS, INC
GASSER ET AL., NATURE LETTERS, 2005
GAZDAR, ONCOGENE, vol. 28, August 2009 (2009-08-01), pages S24 - S31
HAMANEHYU, CANCER INFORM., vol. 21, 2022
JACKOLA, D. R.RUGER, J. K.MILLER, R. A.: "Age-associated changes in human T cell phenotype and function", AGING CLIN. EXP. RES., vol. 6, 1994, pages 25 - 34
LEFRANC, M.P. ET AL., NUCL. ACIDS RES., vol. 27, 1999, pages 209 - 212
LEFRANC. M.P. ET AL., BIOMOLECULES, vol. 4, no. 4, December 2014 (2014-12-01), pages 1102 - 1139
MAKABE ET AL., J. BIOL. CHEM., vol. 283, 2007, pages 1156 - 1166
MARASCO ET AL., PROC. NATL. ACAD. SCI. USA, vol. 90, 1993, pages 7889 - 7893
MUÑOZ, D. P. ET AL.: "Targetable mechanisms driving immunoevasion of persistent senescent cells link chemotherapy-resistant cancer to aging", JCI INSIGHT, vol. 5, 2019, pages 124716
NATURE, vol. 361, 1993, pages 186 - 87
P. TIJSSEN: "The McGraw-Hill Dictionary of Chemical Terms", 1985, ELSEVIER SCIENCE PUBLISHERS
POWELL ET AL.: "Compendium of excipients for parenteral formulations", PDA J PHARM SCI TECHNOL., vol. 52, 1998, pages 238 - 311, XP009119027
RIBAS ET AL., CANCER IMMUNOL RES, vol. 4, no. 3, 2016, pages 194 - 203
ROBERTSON ET AL., CELL, vol. 171, no. 3, 2017, pages 540 - 556
RODIER, F. ET AL.: "Persistent DNA damage signaling triggers senescence-associated inflammatory cytokine secretion", NAT. CELL BIOL., vol. 11, 2009, pages 973 - 979
RUIZ, M., NUCL. ACIDS RES., vol. 28, 2000, pages 219 - 221
RUSCETTI, M. ET AL.: "NK cell-mediated cytotoxicity contributes to tumor control by a cytostatic drug combination", SCIENCE, vol. 362, 2018, pages 1416 - 1422
SAGIV, A. ET AL.: "NKG2D ligands mediate immuno surveillance of senescent cells", AGING, vol. 8, 2016, pages 328 - 344
SAMBROOK ET AL.: "Molecular Cloning: A Laboratory Manual", 1989, COLD SPRING HARBOR LABORATORY PRESS
SU ET AL., BIOINFORMATICS, vol. 36, no. 2, 2019, pages 339 - 346
TAO KELONG ET AL: "Development of NKG2D-based chimeric antigen receptor-T cells for gastric cancer treatment", CANCER CHEMOTHERAPY AND PHARMACOLOGY, SPRINGER VERLAG , BERLIN, DE, vol. 82, no. 5, 21 August 2018 (2018-08-21), pages 815 - 827, XP036609209, ISSN: 0344-5704, [retrieved on 20180821], DOI: 10.1007/S00280-018-3670-0 *
THORNTON, NATURE, vol. 354, 1991, pages 105
WANG W.: "Lyophilization and development of solid protein pharmaceuticals", INT. J. PHARM., vol. 203, no. 1-2, 2000, pages 1 - 60, XP055005330, DOI: 10.1016/S0378-5173(00)00423-3
YAMAGUCHI KOSUKE ET AL: "16-binding protein 2 in lung cancer patients", CANCER SCIENCE, vol. 103, no. 8, 10 July 2012 (2012-07-10), JP, pages 1405 - 1413, XP093229657, ISSN: 1347-9032, DOI: 10.1111/j.1349-7006.2012.02330.x *
YA-TING CHANG ET AL: "Secretome-Based Identification of ULBP2 as a Novel Serum Marker for Pancreatic Cancer Detection", PLOS ONE, vol. 6, no. 5, 1 January 2011 (2011-01-01), pages e20029, XP055036528, ISSN: 1932-6203, DOI: 10.1371/journal.pone.0020029 *
YOST ET AL., NATURE MED, vol. 25, 2019, pages 1251 - 1259

Also Published As

Publication number Publication date
WO2025085512A9 (fr) 2025-06-05

Similar Documents

Publication Publication Date Title
US20220332823A1 (en) Methods of treating cancer
JP7503887B2 (ja) がんを監視及び治療するための方法
JP2022116095A (ja) Ｃｄ４７及びｅｇｆｒの二重標的化による癌治療
JP2017501167A (ja) Ｏｘ４０結合アゴニスト及びｐｄ−１軸結合アンタゴニストを含む併用療法
JP6649941B2 (ja) Ｆｓｔｌ１を利用した抗がん剤・転移抑制剤およびその併用剤
TW201625270A (zh) 用於治療贅瘤形成的治療組合及方法
JP2020531854A (ja) 癌の治療および診断のためのｔｉｍ−３アンタゴニスト
WO2020185739A1 (fr) Anticorps anti-icos pour le traitement du cancer
WO2021089704A1 (fr) Inhibition combinée de pd-1, tgfb et tigit pour le traitement du cancer
JP2020529422A (ja) 急性骨髄性白血病を治療するためのｆｌｔ３ ｃａｒ−ｔ細胞及びｆｌｔ３阻害剤の使用
TW201642897A (zh) Ｈｅｒ２結合劑治療
WO2016161415A2 (fr) Protéines de tnfrsf14/hvem et leurs procédés d'utilisation
JP7455749B2 (ja) 頭頸部癌の処置
KR20200070341A (ko) 항-cd47 및 항-pd-l1로 난소암의 치료
US20240199732A1 (en) Anti-IL-27 Antibodies and Uses Thereof
CA3213110A1 (fr) Methodes de traitement du cancer
JP2023502023A (ja) Ｃｄ２００受容体アンタゴニスト結合分子
JP2022521541A (ja) 免疫機能を増強するための細胞、組成物、及び方法
WO2025085512A9 (fr) Polythérapies avec des anticorps ciblant ulbp2 pour traiter des cancers
WO2020109627A1 (fr) Polythérapie anti-neuropiline-1 et anti-mort cellulaire programmée 1 pour le traitement du cancer
WO2024120484A1 (fr) Anticorps dirigés contree edil3 et leurs méthodes d'utilisation
WO2020086943A1 (fr) Procédés de traitement du cancer
WO2020086802A1 (fr) Procédés et compositions pour le traitement du cancer et de maladies infectieuses
HK40084229A (en) Treatment of cancer with dual targeting of cd47 and egfr
CN120500625A (zh) 治疗肿瘤的方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24802349

Country of ref document: EP

Kind code of ref document: A1