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WO2024163723A9 - Molécules se liant à des polypeptides de follistatine - Google Patents

Molécules se liant à des polypeptides de follistatine Download PDF

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Publication number
WO2024163723A9
WO2024163723A9 PCT/US2024/013973 US2024013973W WO2024163723A9 WO 2024163723 A9 WO2024163723 A9 WO 2024163723A9 US 2024013973 W US2024013973 W US 2024013973W WO 2024163723 A9 WO2024163723 A9 WO 2024163723A9
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WO
WIPO (PCT)
Prior art keywords
seq
amino acid
set forth
antibody
chain variable
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.)
Ceased
Application number
PCT/US2024/013973
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English (en)
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WO2024163723A2 (fr
WO2024163723A3 (fr
Inventor
Dimiter Stanchev Dimitrov
John W. Mellors
Ronald BUCKANOVICH
Zehua Sun
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.)
University of Pittsburgh
Original Assignee
University of Pittsburgh
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Publication date
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Priority to EP24751006.8A priority Critical patent/EP4658312A2/fr
Priority to AU2024213820A priority patent/AU2024213820A1/en
Publication of WO2024163723A2 publication Critical patent/WO2024163723A2/fr
Publication of WO2024163723A3 publication Critical patent/WO2024163723A3/fr
Publication of WO2024163723A9 publication Critical patent/WO2024163723A9/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/10—Cellular immunotherapy characterised by the cell type used
    • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
    • A61K40/31—Chimeric antigen receptors [CAR]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A61K40/4238—Regulators of development
    • 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
    • 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
    • C07K16/2809—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 the T-cell receptor (TcR)-CD3 complex
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
    • C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • BACKGROUND 1 relates to methods and materials involved in binding a molecule (e.g., an antibody, a fragment of an antibody, an antibody domain, a chimeric antigen receptor (CAR), a cell engager, or an antibody-drug conjugate (ADC)) to a follistatin polypeptide.
  • a molecule e.g., an antibody, a fragment of an antibody, an antibody domain, a chimeric antigen receptor (CAR), a cell engager, or an antibody-drug conjugate (ADC)
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, cell engagers, or ADCs
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, cell engagers, or ADCs
  • FST farnesoid follistatin
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, or cell engagers
  • FST Background Information Follistatin
  • activin-binding protein is an extracellular glycoprotein (with a size of about 35 kD of the most dominant form) encoded in humans Attorney Docket No.48881-0047WO1 / 05958 by the FST gene. FST was identified as an inhibitor of pituitary follicle stimulating hormone secretion.
  • FST primarily functions in binding and neutralization of members of the TGF-? superfamily, with a particular focus on activin, which is involved in proliferation, differentiation, and apoptosis of a number of cell types. FST is involved in tumorigenesis, metastasis, and angiogenesis of solid tumors through its interaction with activin. High serum levels of follistatin polypeptides were reported in patients with ovarian cancer, indicating that follistatin polypeptides can be a potential tumor marker for ovarian cancer diagnosis. Thus, follistatin polypeptides are a potential target for treating different types of cancers.
  • This document provides methods and materials involved in binding a molecule (e.g., an antibody, an antigen binding fragment, an antibody domain, a CAR, a cell engager, or an ADC) to a follistatin polypeptide.
  • a molecule e.g., an antibody, an antigen binding fragment, an antibody domain, a CAR, a cell engager, or an ADC
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, cell engagers, or ADCs
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, cell engagers, or ADCs
  • this document provides methods and materials for using one or more binders described herein in combination with one or more chemotherapeutic agents to treat a mammal (e.g., a human) having cancer (e.g., ovarian cancer, lung cancer, and/or prostate cancer).
  • cancer e.g., ovarian cancer, lung cancer, and/or prostate cancer.
  • cells e.g., host cells
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, or cell engagers
  • this document provides methods and materials for using one or more of such cells in combination with one or more chemotherapeutic agents to treat a mammal (e.g., a human) having cancer (e.g., ovarian cancer, lung cancer, and/or prostate cancer).
  • a mammal e.g., a human
  • cancer e.g., ovarian cancer, lung cancer, and/or prostate cancer.
  • binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more CARs, one or more cell engagers, and/or one or more ADCs
  • binders can be designed to have the ability to bind to a follistatin polypeptide.
  • a binder e.g., an antibody, an antigen binding fragment, an antibody domain, a CAR, a cell engager, or an ADC
  • a polypeptide comprising, consisting essentially of, or consisting of the amino acid sequence of a human follistatin polypeptide as set forth in SEQ ID NO:113 and/or SEQ ID NO:115 (see, e.g., Example 2).
  • anti-follistatin binders e.g., Clones #1-#14
  • follistatin polypeptides e.g., human follistatin polypeptides. See, e.g., Figures 4-6.
  • anti-follistatin binder described herein e.g., any one of Clones #1-#14
  • a chemotherapeutic agent e.g., taxol or cisplatin
  • anti-follistatin binders provided herein can be used to treat cancer (e.g., ovarian cancer, lung cancer, and/or prostate cancer).
  • a single set of three CDRs of an antibody domain (e.g., a VH domain) provided herein (e.g., SEQ ID NOs:1-3; SEQ ID NOs:9-11; SEQ ID NOs:17-19; SEQ ID NOs:25-27; SEQ ID NOs:33-35; SEQ ID NOs:41-43; SEQ ID NOs:49-51; SEQ ID NOs:57-59; SEQ ID NOs:65-67; SEQ ID NOs:73-75; SEQ ID NOs:81-83; SEQ ID NOs:89-91; SEQ ID NOs:97-99; or SEQ ID NOs:105-107) can be engineered into a CAR to create CAR + cells (e.g., CAR + T cells, CAR + stem cells such as CAR + induced pluripotent stem cells, or CAR + NK cells) having the ability to target follistatin + cells (e.g., follistatin + tumor cells and
  • binders e.g., one or more antibodies, one or more antigen binding fragments, and/or one or more antibody domains
  • ADCs such as full antibody-drug conjugates, Fab-drug conjugates, scFv-drug conjugates, and/or antibody domain-drug conjugates can be designed to include an appropriate binder provided herein to create the conjugate.
  • Such conjugates can be used to deliver the drug payload to target cells such as cancer cells (e.g., follistatin + cancer cells) or cancer vasculature (e.g., follistatin + cancer vasculature).
  • target cells such as cancer cells (e.g., follistatin + cancer cells) or cancer vasculature (e.g., follistatin + cancer vasculature).
  • binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a mammal e.g., a human having cancer.
  • a mammal e.g., a human having cancer (e.g., a follistatin + cancer) can be administered a composition comprising one or more binders (e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs) described herein to reduce the number of cancer cells within the mammal, to induce ADCC against cancer cells within the mammal, and/or to increase the survival duration of the mammal from cancer (e.g., ovarian cancer, lung cancer, and/or prostate cancer).
  • binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • cells e.g., host cells
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, or cell engagers
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, or cell engagers
  • cells such as T cells (e.g., CTLs), stem cells (e.g., induced pluripotent stem cells), or NK cells can be engineered to express one or more CARs having the ability to bind to a follistatin polypeptide.
  • Such cells can be used to treat cancer (e.g., ovarian cancer, lung cancer, and/or prostate cancer).
  • cancer e.g., ovarian cancer, lung cancer, and/or prostate cancer.
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a sample e.g., a biological sample such tumor biopsy
  • a mammal e.g., a human
  • follistatin + cells e.g., follistatin + cancer cells.
  • follistatin + cancer cells detection of follistatin + cancer cells within a mammal can allow clinicians, health professionals, and patients to select an appropriate anti-cancer treatment that targets the follistatin + cancer cells.
  • treatments that target the follistatin + cancer cells can include administration of one or more of the binders described herein having the ability to bind to a follistatin polypeptide and/or administration of one or more cells (e.g., follistatin-specific CAR + T cells or NK cells) designed to express a binder described herein.
  • one aspect of this document features an antibody comprising (or consisting essentially of, or consisting of) (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two, or three amino acid additions, deletions, or substitutions); (ii) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two,
  • the antibody can comprise the ability to bind to SEQ ID NO:113 and/or SEQ ID NO:115.
  • the antibody can comprise the heavy chain variable domain or region of (i).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8.
  • the antibody can comprise the heavy chain variable domain or region of (ii).
  • the heavy Attorney Docket No.48881-0047WO1 / 05958 chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16.
  • the antibody can comprise the heavy chain variable domain or region of (iii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24.
  • the antibody can comprise the heavy chain variable domain or region of (iv).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32.
  • the antibody can comprise the heavy chain variable domain or region of (v).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:40.
  • the antibody can comprise the heavy chain variable domain or region of (vi).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:48.
  • the antibody can comprise the heavy chain variable domain or region of (vii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:56.
  • the antibody can comprise the heavy chain variable domain or region of (viii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:64.
  • the antibody can comprise the heavy chain variable domain or region of (ix).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:72.
  • the antibody can comprise the heavy chain variable domain or region of (x).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:80.
  • the antibody can comprise the heavy chain variable domain or region of (xi).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:88.
  • the antibody can comprise the heavy chain variable domain or region of (xii).
  • the heavy chain variable domain or region can Attorney Docket No.48881-0047WO1 / 05958 comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:96.
  • the antibody can comprise the heavy chain variable domain or region of (xiii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:104.
  • the antibody can comprise the heavy chain variable domain or region of (xiv).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:112.
  • the antibody can be a monoclonal antibody.
  • the antibody can be an scFv antibody. This paragraph can be referred to as first aspect paragraph.
  • this document features an antigen binding fragment comprising (or consisting essentially of, or consisting of): (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two, or three amino acid additions, deletions, or substitutions); (ii) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one,
  • the antigen binding fragment can comprise the ability to bind to SEQ ID NO:113 and/or SEQ ID NO:115.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (i).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (ii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (iii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the Attorney Docket No.48881-0047WO1 / 05958 amino acid sequence set forth in SEQ ID NO:24.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (iv).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (v).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:40.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (vi).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:48.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (vii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:56.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (viii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:64.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (ix).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:72.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (x).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:80.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (xi).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:88.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (xii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:96.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (xiii).
  • the heavy chain variable domain or region can comprise an amino acid sequence Attorney Docket No.48881-0047WO1 / 05958 having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:104.
  • the antigen binding fragment can comprise the heavy chain variable domain or region of (xiv).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:112.
  • the antigen binding fragment can be monoclonal.
  • the antigen binding fragment can be a Fab. This paragraph can be referred to as second aspect paragraph.
  • this document features an antibody domain comprising (or consisting essentially of, or consisting of): (i) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:1 (or SEQ ID NO:1 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:2 (or SEQ ID NO:2 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:3 (or SEQ ID NO:3 with one, two, or three amino acid additions, deletions, or substitutions); (ii) a heavy chain variable domain or region comprising the amino acid sequences set forth in SEQ ID NO:9 (or SEQ ID NO:9 with one, two, or three amino acid additions, deletions, or substitutions), SEQ ID NO:10 (or SEQ ID NO:10 with one, two, or three amino acid additions, deletions, or substitutions), and SEQ ID NO:11 (or SEQ ID NO:11 with one, two,
  • the antibody domain can comprise the ability to bind to SEQ ID NO:113 and/or SEQ ID NO:115.
  • the antibody domain can comprise the heavy chain variable domain or region of (i).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8.
  • the antibody domain can comprise the heavy chain variable domain or region of (ii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16.
  • the antibody domain can comprise the heavy chain variable domain or region of (iii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24.
  • the antibody domain can comprise the heavy chain variable domain or region of (iv).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32.
  • the antibody domain can comprise the heavy chain variable domain or region of (v).
  • the Attorney Docket No.48881-0047WO1 / 05958 heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:40.
  • the antibody domain can comprise the heavy chain variable domain or region of (vi).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:48.
  • the antibody domain can comprise the heavy chain variable domain or region of (vii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:56.
  • the antibody domain can comprise the heavy chain variable domain or region of (viii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:64.
  • the antibody domain can comprise the heavy chain variable domain or region of (ix).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:72.
  • the antibody domain can comprise the heavy chain variable domain or region of (x).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:80.
  • the antibody domain can comprise the heavy chain variable domain or region of (xi).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:88.
  • the antibody domain can comprise the heavy chain variable domain or region of (xii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:96.
  • the antibody domain can comprise the heavy chain variable domain or region of (xiii).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:104.
  • the antibody domain can comprise the heavy chain variable domain or region of (xiv).
  • the heavy chain variable domain or region can comprise an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:112.
  • the Attorney Docket No.48881-0047WO1 / 05958 antibody domain can be monoclonal.
  • the antibody domain can be a VH domain. This paragraph can be referred to as third aspect paragraph.
  • this document features a chimeric antigen receptor comprising (or consisting essentially of, or consisting of) an antigen binding domain, a hinge, a transmembrane domain, and one or more signaling domains, wherein the antigen binding domain comprises (or consists essentially of, or consists of) any antibody, any antigen- binding fragment, or any antibody domain of any of first aspect paragraph, second aspect paragraph, or third aspect paragraph.
  • the antigen binding domain can comprise a VH domain having the ability to bind to a follistatin polypeptide.
  • the hinge can comprise a hinge set forth in Example 23.
  • the transmembrane domain can comprise a transmembrane domain set forth in Example 24.
  • the chimeric antigen receptor can comprise one or more signaling domains set forth in Example 25.
  • This paragraph can be referred to as fourth aspect paragraph.
  • this document features a cell comprising (or consisting essentially of, or consisting of) any chimeric antigen receptor of the preceding paragraph.
  • the cell can be a T cell, a stem cell, or an NK cell.
  • this document features a cell engager comprising (or consisting essentially of, or consisting of) a first antigen binding domain, a linker, and a second antigen binding domain, wherein the first antigen binding domain comprises (or consists essentially of, or consists of) any antibody, any antigen-binding fragment, or any antibody domain of any of first aspect paragraph, second aspect paragraph, or third aspect paragraph.
  • the first antigen binding domain can comprise a VH domain having the ability to bind to a follistatin polypeptide.
  • the first antigen binding domain can be an IgG having the ability to bind to a follistatin polypeptide.
  • the linker can comprise a linker set forth in Example 20 or Example 23.
  • the second antigen binding domain can bind to a polypeptide expressed on the surface of T cells.
  • the polypeptide expressed on the surface of T cells can be a CD3 polypeptide.
  • the second antigen binding domain can be an antigen binding domain set forth in Example 28.
  • the second antigen binding domain can bind to a polypeptide expressed on the surface of NK cells.
  • the polypeptide expressed on the surface of NK cells can be a CD16a, NKG2A, NKG2D, NKp30, Attorney Docket No.48881-0047WO1 / 05958 NKp44, or NKp46 polypeptide.
  • the second antigen binding domain can be an antigen binding domain set forth in Example 29.
  • the cell engager can comprise a third antigen binding domain.
  • the third antigen binding domain can bind to a polypeptide expressed on the surface of NK cells.
  • the polypeptide expressed on the surface of NK cells can be a CD16a, NKG2A, NKG2D, NKp30, NKp44, or NKp46 polypeptide.
  • the third antigen binding domain can be an antigen binding domain set forth in Example 29. This paragraph can be referred to as fifth aspect paragraph.
  • this document features a nucleic acid comprising (or consisting essentially of, or consisting of) a nucleic acid sequence encoding at least part of the antibody, the antigen-binding fragment, or the antibody domain of any of first aspect paragraph, second aspect paragraph, or third aspect paragraph.
  • the nucleic acid sequence can encode the heavy chain variable domain or region of any one of the (i)- (xiv) of first aspect paragraph.
  • the nucleic acid can be a viral vector.
  • the nucleic acid can be a phagemid.
  • this document features a nucleic acid comprising (or consisting essentially of, or consisting of) a nucleic acid sequence encoding any chimeric antigen receptor of fourth aspect paragraph or any cell engager of fifth aspect paragraph.
  • the nucleic acid can be a viral vector.
  • the nucleic acid can be a phagemid.
  • this document features a host cell comprising (or consisting essentially of, or consisting of) any nucleic acid of the preceding paragraph.
  • this document features a host cell that expresses any chimeric antigen receptor of fourth aspect paragraph or any cell engager of fifth aspect paragraph.
  • the host cell can be a T cell, stem cell, or NK cell.
  • This paragraph can be referred to as seventh aspect paragraph.
  • this document features an antibody-drug conjugate (ADC) comprising (or consisting essentially of, or consisting of) an antigen binging domain covalently linked to a drug, wherein the antigen binging domain comprises an antibody, an antigen binding fragment, or an antibody domain of any of first aspect paragraph, second aspect paragraph, or third aspect paragraph.
  • ADC antibody-drug conjugate
  • the antigen binding domain can Attorney Docket No.48881-0047WO1 / 05958 comprise a VH domain having the ability to bind to a follistatin polypeptide.
  • the drug can be selected from the group consisting of auristatins, mertansine, or pyrrolobenzodiazepine (PBD) dimers.
  • PBD pyrrolobenzodiazepine
  • This paragraph can be referred to as eighth aspect paragraph.
  • this document features a composition comprising (or consisting essentially of, or consisting of) an antibody, an antigen binding fragment, or an antibody domain of any of first aspect paragraph, second aspect paragraph, or third aspect paragraph.
  • the composition can comprise any antibody of first aspect paragraph.
  • the composition can comprise any antigen binding fragment of second aspect paragraph.
  • composition can comprise any antibody domain of third aspect paragraph.
  • this document features a composition comprising (or consisting essentially of, or consisting of) any cell engager of fifth aspect paragraph.
  • this document features a composition comprising (or consisting essentially of, or consisting of) any cell of sixth aspect paragraph or seventh aspect paragraph.
  • this document features a composition comprising (or consisting essentially of, or consisting of) any ADC of any one of eighth aspect paragraph.
  • the composition can comprise a checkpoint inhibitor.
  • the checkpoint inhibitor can be selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, JTX- 4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostarlimab, INCMGA00012, AMP-224, AMP-514, avelumab, durvalumab, atezolizumab, KN035, CK-301, AUNP12, CA-170, BMS-986189, and ipilimumab.
  • this document features a method of treating a mammal having cancer.
  • the method comprises (or consists essentially of, or consists of) administering, to the mammal, a composition of any of the preceding four paragraphs.
  • the mammal can be a human.
  • the cancer can be a follistatin + cancer.
  • the follistatin + cancer can be selected from the group consisting of follistatin + ovarian cancer, follistatin + lung cancer, and follistatin + prostate cancer.
  • the number of cancer cells within the mammal can be reduced following the administering step.
  • Attorney Docket No.48881-0047WO1 / 05958 In another aspect, this document features a method of treating a mammal having cancer.
  • the method comprises (or consists essentially of, or consists of) (a) administering, to the mammal, a composition of any of the preceding four paragraphs referred to in the preceding paragraph, and (b) administering, to the mammal, a composition comprising a checkpoint inhibitor.
  • the mammal can be a human.
  • the cancer can be a follistatin + cancer.
  • the follistatin + cancer can be selected from the group consisting of follistatin + ovarian cancer, follistatin + lung cancer, and follistatin + prostate cancer.
  • the checkpoint inhibitor can be selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostarlimab, INCMGA00012, AMP-224, AMP- 514, avelumab, durvalumab, atezolizumab, KN035, CK-301, AUNP12, CA-170, BMS- 986189, and ipilimumab.
  • the number of cancer cells within the mammal can be reduced following the administering steps (a) and (b).
  • this document features a method for binding a binding molecule to a follistatin polypeptide.
  • the method comprises (or consists essentially of, or consists of) contacting the follistatin polypeptide with an antibody, an antigen binding fragment, or an antibody domain of any of first aspect paragraph, second aspect paragraph, or third aspect paragraph.
  • the contacting can be performed in vitro.
  • the contacting can be performed in vivo.
  • the contacting can be performed within a mammal by administering the antibody, the antigen binding fragment, or the antibody domain to the mammal.
  • the mammal can be a human.
  • this document features a method for binding a binding molecule to a follistatin polypeptide.
  • the method comprises (or consists essentially of, or consists of) contacting the follistatin polypeptide with any chimeric antigen receptor of fourth aspect paragraph, any cell engager of fifth aspect paragraph, or any ADC of eighth aspect paragraph.
  • the contacting can be performed in vitro.
  • the contacting can be performed in vivo.
  • the contacting can be performed within a mammal by administering the chimeric antigen receptor, the cell engager, or the ADC to the mammal.
  • the mammal can be a human.
  • Attorney Docket No.48881-0047WO1 / 05958 Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains.
  • a promotor sequence (e.g., a CMV immediate early promotor sequence) can be followed by a signal peptide sequence (e.g., a GM-CSF signal peptide sequence), followed by a VH domain provided herein (e.g., a VH domain designed to include a set of three CDRs such as CDR1, CDR2, and CDR3 of a VH domain provided herein, for example, SEQ ID NOs:1-3; SEQ ID NOs:9-11; SEQ ID NOs:17-19; SEQ ID NOs:25-27; SEQ ID NOs:33-35; SEQ ID NOs:41-43; SEQ ID NOs:49-51; SEQ ID NOs:57-59; SEQ ID NOs:65-67; SEQ ID NOs:73-75; SEQ ID NOs:81-83; SEQ ID NOs:89-91; SEQ ID Attorney Docket No.48881-0047WO1 / 05958 NOs:97-99; or S
  • FIG. 3 is a schematic of an exemplary BiTE designed using CDR1, CDR2, and CDR3 of a heavy chain and CDR1, CDR2, and CDR3 of a light chain in an Ig format (e.g., an IgG1 format).
  • a humanized anti-CD3 scFv e.g., an gOKT3-7 scFv set forth in U.S. Patent No.6,750,325
  • a linker e.g., a (G4S)3 linker; SEQ ID NO:139.
  • Figure 4 is a graph plotting the binding of the indicated VH domain binders to a recombinant human follistatin polypeptide as determined by ELISA. 200 ng of a recombinant human follistatin polypeptide was coated onto immune-absorb plates, and the indicated concentrations of the indicated binders were added. After incubation and washing, horseradish peroxidase (HRP)-conjugated mouse anti-FLAG tag antibody (A8592, Sigma–Aldrich) was used to detect VH binding.
  • Figure 5 is a graph plotting the binding of the indicated binders in an Fc format to a recombinant human follistatin polypeptide as determined by ELISA.
  • HRP horseradish peroxidase
  • FIG. 6 is a graph plotting the binding of the indicated VH domain binders to BSA as determined by ELISA.
  • FIG. 8 contains graphs plotting nuclear DNA content in ovarian cancer cells treated with or without chemotherapy (taxol) in the presence or absence of an anti- follistatin VH clone (FST3G9).
  • taxol increases nuclear content leading to mitotic catastrophe. Addition of an anti-follistatin VH clone (FST3G9) significantly enhanced this effect.
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, cell engagers, and ADCs
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, cell engagers, and ADCs
  • bind e.g., specifically bind
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • the document provides binders (e.g., antibodies, antigen binding fragments, antibody domains, CARs, cell engagers, and ADCs) that bind (e.g., specifically bind) to a polypeptide comprising, consisting essentially of, or consisting of the amino acid set forth in SEQ ID NO:113 and/or SEQ ID NO:115 (see, e.g., Example 2).
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, cell engagers, and ADCs
  • antibody as used herein includes polyclonal antibodies, monoclonal antibodies, recombinant antibodies, humanized antibodies, human antibodies, chimeric antibodies, multi-specific antibodies (e.g., bispecific antibodies) formed from at least two antibodies, diabodies, single-chain variable fragment antibodies (e.g., scFv antibodies), and tandem single-chain variable fragments antibody (e.g., taFv).
  • a diabody can include two chains, each having a heavy chain variable domain and a light chain variable domain, either from the same or from different antibodies (see, e.g., Hornig and Desirber-Schwarz, Methods Mol.
  • the two variable regions can be connected by a polypeptide linker (e.g., a polypeptide linker having five to ten residues in length or a polypeptide linker as set forth in Example 20).
  • a polypeptide linker e.g., a polypeptide linker having five to ten residues in length or a polypeptide linker as set forth in Example 20.
  • an interdomain disulfide bond can be present in one or both of the heavy chain variable domain and light chain variable domain pairs of the diabody.
  • a scFv is a single-chain polypeptide antibody in which the heavy chain variable domain and the light chain variable domain are directly connected or connected via a polypeptide linker (e.g., a polypeptide linker having eight to 18 residues in length or a polypeptide linker as set forth in Example 20). See, also, Chen et al., Adv. Drug Deliv. Rev., 65(10):1357-1369 (2013).
  • a scFv can be designed to have an orientation with the heavy chain variable domain being followed by the light chain variable domain or can be designed to have an orientation with the light chain variable domain being followed by the heavy chain variable domain. In both cases, the optional linker can be located between the two domains.
  • Examples of scFv structures of scFvs provided herein include, without limitation, those structures set forth in Example 19. Attorney Docket No.48881-0047WO1 / 05958
  • An antibody provided herein can include the CDRs as described herein (e.g., as described in Table 43) and can be configured to be a human antibody, a humanized antibody, or a chimeric antibody.
  • an antibody provided herein can include the CDRs as described herein (e.g., as described in Table 43) and can be a monoclonal antibody.
  • an antibody provided herein can include the CDRs as described herein (e.g., as described in Table 43) and can be configured as a scFv antibody.
  • antigen binding fragment refers to a fragment of an antibody (e.g., a fragment of a humanized antibody, a fragment of a human antibody, or a fragment of a chimeric antibody) having the ability to bind to an antigen.
  • antigen binding fragments include, without limitation, Fab, Fab’, or F(ab’)2 antigen binding fragments.
  • An antigen binding fragment provided herein can include the CDRs as described herein (e.g., as described in Table 43) and can be configured to be a human antigen binding fragment, a humanized antigen binding fragment, or a chimeric antigen binding fragment.
  • an antigen binding fragment provided herein can include the CDRs as described herein (e.g., as described in Table 43) and can be a monoclonal antigen binding fragment.
  • an antigen binding fragment provided herein can include the CDRs as described herein (e.g., as described in Table 43) and can be configured as a Fab antibody.
  • a Fab antibody can include a partial hinge sequence (e.g., EPKSCDKT (SEQ ID NO:228)) for disulfide bonding between heavy and light chains of the Fab.
  • antibody domain refers to a domain of an antibody such as a heavy chain variable domain (VH domain) or a light chain variable domain (VL domain) in the absence of one or more other domains of an antibody.
  • an antibody domain can be a single antibody domain (e.g., a VH domain or a VL domain) having the ability to bind to an antigen.
  • an antibody domain provided herein can include the CDRs as described herein (e.g., as described in Table 43) and can be engineered as a single VH domain or a single VL domain.
  • Examples of VH domains provided herein include, without limitation, those structures set forth in Examples 3-16.
  • An anti-follistatin antibody, anti-follistatin antigen binding fragment, or anti- follistatin antibody domain provided herein can be of the IgA-, IgD-, IgE-, IgG-, or IgM- type, including IgG- or IgM-types such as, without limitation, IgG1-, IgG2-, IgG3-, IgG4-, IgM1-, and IgM2-types.
  • an antibody provided herein e.g., an anti- follistatin antibody
  • an antigen binding fragment provided herein e.g., an anti-follistatin antibody fragment
  • an antibody provided herein can be a fully intact antibody having the structure set forth in Example 18.
  • an antibody domain provided herein e.g., an anti-follistatin antibody domain
  • an anti-follistatin antibody, anti-follistatin antigen binding fragment, or anti-follistatin antibody domain provided herein can be fully human.
  • an anti-follistatin antibody, anti-follistatin antigen binding fragment, or anti- follistatin antibody domain provided herein can have a low risk for inducing immunogenicity within a human.
  • chimeric antigen receptor refers to a chimeric polypeptide that is designed to include an optional signal peptide, an antigen binding domain, an optional hinge, a transmembrane domain, and one or more intracellular signaling domains.
  • the antigen binding domain of a CAR provided herein can be designed to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a CAR provided herein can be designed to include the components of an antibody, antigen binding fragment, and/or antibody domain described herein (e.g., a combination of CDRs) as an antigen binding domain provided that that antigen binding domain has the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a CAR provided herein can be designed to include an antigen binding domain that includes a single set of three CDRs (e.g., a CDR1, CDR2, and CDR3) of an antibody domain (e.g., a VH domain) provided herein (e.g., Attorney Docket No.48881-0047WO1 / 05958 SEQ ID NOs:1-3; SEQ ID NOs:9-11; SEQ ID NOs:17-19; SEQ ID NOs:25-27; SEQ ID NOs:33-35; SEQ ID NOs:41-43; SEQ ID NOs:49-51; SEQ ID NOs:57-59; SEQ ID NOs:65-67; SEQ ID NOs:73-75; SEQ ID NOs:81-83; SEQ ID NOs:89-91; SEQ ID NOs:97-99; or SEQ ID NOs:105-107).
  • a single set of three CDRs e.g., a CDR1, CDR2, and C
  • an antigen binding domain of a CAR targeting a follistatin polypeptide can be designed to include a VH domain described herein or a scFv antibody described herein.
  • Examples of CAR structures that can be used to make a CAR provided herein include, without limitation, those set forth in Figures 1 and 2 and Examples 21 and 26.
  • a CAR provided herein can be designed to include a signal peptide. Any appropriate signal peptide can be used to design a CAR described herein.
  • Examples of signal peptide that can be used to make a CAR described herein include without limitation, a human IGKV1-39-derived signal peptide, IGKV1-16, IGKV1-33, IGKV3- 11, IGKV4-1, or IGKV6-21.
  • a CAR provided herein can be designed to include a signal peptide that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 22.
  • a CAR provided herein can be designed to include a hinge. Any appropriate hinge can be used to design a CAR described herein.
  • hinges that can be used to make a CAR described herein include, without limitation, Ig-derived hinges (e.g., an IgG1-derived hinge, an IgG2-derived hinge, or an IgG4-derived hinge), Ig-derived hinges containing a CD2 domain and a CD3 domain, Ig-derived hinges containing a CD2 domain and lacking a CD3 domain, Ig-derived hinges containing a CD3 domain and lacking a CD2 domain, Ig-derived hinges lacking a CD2 domain and Attorney Docket No.48881-0047WO1 / 05958 lacking a CD3 domain, CD8?-derived hinges, CD28-derived hinges, and CD3?-derived hinges.
  • a CAR provided herein can be designed to include a hinge of any appropriate length.
  • a CAR provided herein can be designed to include a hinge that is from about 3 to about 75 (e.g., from about 3 to about 65, from about 3 to about 50, from about 5 to about 75, from about 10 to about 75, from about 5 to about 50, from about 10 to about 50, from about 10 to about 40, or from about 10 to about 30) amino acid residues in length.
  • a linker sequence can be used as a hinge to make a CAR described herein.
  • any one of the linker sequences set forth in Example 20 can be used as a hinge of a CAR described herein.
  • a CAR provided herein can be designed to include a hinge that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 20 or Example 23. In some cases, a CAR provided herein can be designed to include a hinge that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 20 or Example 23 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof.
  • a CAR provided herein can be designed to include a hinge that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 20 or Example 23 with two or less, three or less, four or less, five or less, six or less, seven or less, eight or less, nine or less, or ten or less amino acid deletions, additions, substitutions, or combinations thereof.
  • a CAR provided herein can be designed to include any appropriate transmembrane domain.
  • the transmembrane domain of a CAR provided herein can be, without limitation, a CD3? transmembrane domain, a CD4 transmembrane domain, a CD8?
  • a CAR provided herein can be designed to include a transmembrane domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 24.
  • a CAR provided herein can be designed to include a transmembrane domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 24 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, Attorney Docket No.48881-0047WO1 / 05958 additions, substitutions, or combinations thereof.
  • a CAR provided herein can be designed to include a transmembrane domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 24 with two or less, three or less, four or less, five or less, six or less, seven or less, eight or less, nine or less, or ten or less amino acid deletions, additions, substitutions, or combinations thereof.
  • a CAR provided herein can be designed to include one or more intracellular signaling domains.
  • a CAR provided herein can be designed to include one, two, three, or four intracellular signaling domains. Any appropriate intracellular signaling domain or combination of intracellular signaling domains can be used to make a CAR described herein.
  • intracellular signaling domains examples include, without limitation, CD3? intracellular signaling domains, CD27 intracellular signaling domains, CD28 intracellular signaling domains, OX40 (CD134) intracellular signaling domains, 4-1BB (CD137) intracellular signaling domains, CD278 intracellular signaling domains, DAP10 intracellular signaling domains, and DAP12 intracellular signaling domains.
  • a CAR described herein can be designed to be a first generation CAR having a CD3? intracellular signaling domain.
  • a CAR described herein can be designed to be a second generation CAR having a CD28 intracellular signaling domain followed by a CD3? intracellular signaling domain.
  • a CAR described herein can be designed to be a third generation CAR having (a) a CD28 intracellular signaling domain followed by (b) a CD27 intracellular signaling domain, an OX40 intracellular signaling domains, or a 4-1BB intracellular signaling domain followed by (c) a CD3? intracellular signaling domain.
  • a CAR provided herein can be designed to include at least one intracellular signaling domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 25.
  • a CAR provided herein can be designed to include at least one intracellular signaling domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 25 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof, provided that that intracellular signaling domain has at least some activity to activate intracellular signaling.
  • a Attorney Docket No.48881-0047WO1 / 05958 CAR provided herein can be designed to include at least one intracellular signaling domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 25 with two or less, three or less, four or less, five or less, six or less, seven or less, eight or less, nine or less, or ten or less amino acid deletions, additions, substitutions, or combinations thereof, provided that that intracellular signaling domain has at least some activity to activate intracellular signaling.
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:57, SEQ ID NO:58, and SEQ ID NO:59, followed by SEQ ID NO:178, followed by SEQ ID NO:184, followed by SEQ ID NO:192, followed by SEQ ID NO:191 (see, e.g., Example 26).
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:57, SEQ ID NO:58, and SEQ ID NO:59, followed by SEQ ID NO:180, followed by SEQ ID NO:187, followed by SEQ ID NO:194, followed by SEQ ID NO:191 (see, e.g., Example 26).
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:64, followed by a hinge such as a hinge/linker set forth in Example 20 or Example 23 (e.g., an IgG4-derived hinge, a CD8?
  • transmembrane domain such as a transmembrane domain set forth in Example 24 (e.g., a human CD28 transmembrane domain or a CD8? transmembrane domain), followed by one or more intracellular signaling domains such as one or more intracellular signaling domain set forth in Attorney Docket No.48881-0047WO1 / 05958
  • Example 25 e.g., a human 4-1BB intracellular signaling domain followed by a human CD3? intracellular signaling domain.
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:64, followed by SEQ ID NO:178, followed by SEQ ID NO:184, followed by SEQ ID NO:192, followed by SEQ ID NO:191 (see, e.g., Example 26).
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:64, followed by SEQ ID NO:180, followed by SEQ ID NO:187, followed by SEQ ID NO:194, followed by SEQ ID NO:191 (see, e.g., Example 26).
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:65, SEQ ID NO:66, and SEQ ID NO:67, followed by a hinge such as a hinge/linker set forth in Example 20 or Example 23 (e.g., an IgG4-derived hinge, a CD8? hinge, or a linker plus IgG4-derived hinge), followed by a transmembrane domain such as a transmembrane domain set forth in Example 24 (e.g., a human CD28 transmembrane domain or a CD8?
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:65, SEQ ID NO:66, and SEQ ID NO:67, followed by SEQ ID NO:178, followed by SEQ ID NO:184, followed by SEQ ID NO:192, followed by SEQ ID NO:191 (see, e.g., Example 26).
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:65, SEQ ID NO:66, and SEQ ID NO:67, followed by SEQ ID NO:180, followed by SEQ ID NO:187, followed by SEQ ID NO:194, followed by SEQ ID NO:191 (see, e.g., Example 26).
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:72, followed by a hinge such as a hinge/linker set forth in Example 20 or Example 23 (e.g., an IgG4-derived hinge, a CD8?
  • transmembrane domain such as a transmembrane domain set forth in Example 24 (e.g., a human CD28 transmembrane Attorney Docket No.48881-0047WO1 / 05958 domain or a CD8? transmembrane domain), followed by one or more intracellular signaling domains such as one or more intracellular signaling domain set forth in Example 25 (e.g., a human 4-1BB intracellular signaling domain followed by a human CD3? intracellular signaling domain).
  • a transmembrane domain such as a transmembrane domain set forth in Example 24 (e.g., a human CD28 transmembrane Attorney Docket No.48881-0047WO1 / 05958 domain or a CD8? transmembrane domain)
  • intracellular signaling domains such as one or more intracellular signaling domain set forth in Example 25 (e.g., a human 4-1BB intracellular signaling domain followed by a human CD3? intracellular signaling domain
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:72, followed by SEQ ID NO:178, followed by SEQ ID NO:184, followed by SEQ ID NO:192, followed by SEQ ID NO:191 (see, e.g., Example 26).
  • a CAR targeting a follistatin polypeptide can be designed to include a VH domain comprising SEQ ID NO:72, followed by SEQ ID NO:180, followed by SEQ ID NO:187, followed by SEQ ID NO:194, followed by SEQ ID NO:191 (see, e.g., Example 26).
  • cell engager refers to a polypeptide that includes two or more antigen binding domains (e.g., two, three, or four antigen binding domains) and has the ability to link two cells together.
  • cell engagers include, without limitation, BiTEs, BiKEs, and TriKEs.
  • a cell engager provided herein can be designed to include at least one antigen binding domain having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide) and at least one antigen binding domain having the ability to bind to an antigen expressed on the surface of a cell (e.g., a T cell or an NK cell).
  • a cell engager described herein can link a follistatin + cell (e.g., a follistatin + cancer cell) to another cell (e.g., a T cell or an NK cell) via the two or more antigen binding domains of the cell engager.
  • a cell engager structure of cell engagers provided herein includes, without limitation, the structure set forth in Figure 3.
  • the anti-CD3 scFv depicted in Figure 3 can be replaced with a different antigen binding domain having the ability to bind to an antigen expressed on the surface of a cell (e.g., a T cell or an NK cell).
  • a cell engager When a cell engager includes an antigen binding domain having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide) and two or more other antigen binding domains (e.g., two, three, or four other antigen binding domains), each of those other antigen binding domains can bind to different antigens expressed on the surface of different cell types or can bind to different antigens expressed on the surface of the same cell type.
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • two or more other antigen binding domains e.g., two, three, or four other antigen binding domains
  • a TriKE can be designed to have a first Attorney Docket No.48881-0047WO1 / 05958 antigen binding domain having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide), a second antigen binding domain having the ability to bind to a first antigen expressed on the surface of an NK cell (e.g., a CD16 polypeptide such as a CD16a polypeptide), and a third antigen binding domain having the ability to bind to a second antigen expressed on the surface of an NK cell (e.g., an NKG2A polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a second antigen binding domain having the ability to bind to a first antigen expressed on the surface of an NK cell
  • a third antigen binding domain having the ability to bind to a second antigen expressed on the surface of an NK
  • At least one antigen binding domain of a cell engager provided herein can be designed to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a cell engager provided herein can be designed to include the components of an antibody, antigen binding fragment, and/or antibody domain described herein (e.g., a combination of CDRs) as an antigen binding domain provided that that antigen binding domain has the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a cell engager provided herein can be designed to include an antigen binding domain that includes two sets of three CDRs (e.g., CDR1, CDR2, and CDR3 of a heavy chain and CDR1, CDR2, and CDR3 of a light chain) of an antigen binding fragment.
  • CDR1, CDR2, and CDR3 of a heavy chain and CDR1, CDR2, and CDR3 of a light chain of an antigen binding fragment.
  • a cell engager provided herein can be designed to include an antigen binding domain that includes a single set of three CDRs (e.g., a CDR1, CDR2, and CDR3) of an antibody domain (e.g., a VH domain) provided herein (e.g., SEQ ID NOs:1-3; SEQ ID NOs:9-11; SEQ ID NOs:17-19; SEQ ID NOs:25-27; SEQ ID NOs:33- 35; SEQ ID NOs:41-43; SEQ ID NOs:49-51; SEQ ID NOs:57-59; SEQ ID NOs:65-67; SEQ ID NOs:73-75; SEQ ID NOs:81-83; SEQ ID NOs:89-91; SEQ ID NOs:97-99; or SEQ ID NOs:105-107).
  • CDRs e.g., a CDR1, CDR2, and CDR3
  • an antibody domain e.g., a VH domain
  • an antigen binding domain of a cell engager targeting a follistatin polypeptide can be designed to include a VH domain described herein or a scFv/Fab antibody described herein.
  • an antigen binding domain of a CAR described herein that has the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • the cell engager when designing a cell engager such as a BiTE to link a follistatin + cell and a T cell, can include an antigen binding domain having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide) and an antigen binding domain having the ability to bind to a polypeptide expressed on the surface of a T cell.
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • polypeptides expressed on the surface of a T cell that can be targeted by an antigen binding domain of a cell engager provided herein include, without limitation, CD3 polypeptides.
  • antigen binding domains having the ability to bind to a polypeptide expressed on the surface of a T cell that can be used to make a cell engager provided herein (e.g., a BiTE) include, without limitation, anti-CD3 scFvs and anti-CD3 VH domains. Additional examples of amino acid sequences that can be used as antigen binding domains having the ability to bind to a polypeptide expressed on the surface of a T cell (e.g., CD3) are described in U.S. Patent No.6,750,325 (see, e.g., the sequence listing of U.S. Patent No.6,750,325).
  • BiTE structures that can be used to make a BiTE provided herein include, without limitation, those set forth in Examples 30 and 31.
  • a cell engager provided herein can be designed to include an antigen binding domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 28.
  • a cell engager provided herein can be designed to include an antigen binding domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 28 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof, provided that the antigen binding domain has the ability to bind to a polypeptide expressed on the surface of a T cell.
  • a cell engager provided herein can be designed to include an antigen binding domain that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 28 with two or less, three or less, four or less, five or less, Attorney Docket No.48881-0047WO1 / 05958 six or less, seven or less, eight or less, nine or less, or ten or less amino acid deletions, additions, substitutions, or combinations thereof, provided that the antigen binding domain has the ability to bind to a polypeptide expressed on the surface of a T cell.
  • the cell engager can include an antigen binding domain having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide) and one or more (e.g., one, two, or three) antigen binding domains having the ability to bind to a polypeptide expressed on the surface of an NK cell.
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • one or more (e.g., one, two, or three) antigen binding domains having the ability to bind to a polypeptide expressed on the surface of an NK cell.
  • polypeptides expressed on the surface of an NK cell that can be targeted by an antigen binding domain of a cell engager provided herein include, without limitation, CD16 polypeptides (e.g., CD16a polypeptides), NKG2A polypeptides, NKG2D polypeptides, NKp30 polypeptides, NKp44 polypeptides, and NKp46 polypeptides.
  • CD16 polypeptides e.g., CD16a polypeptides
  • NKG2A polypeptides e.g., CD16a polypeptides
  • NKG2D polypeptides e.g., NKG2D polypeptides
  • NKp30 polypeptides e.g., NKp30 polypeptides
  • NKp44 polypeptides e.g., NKp46 polypeptides
  • antigen binding domains having the ability to bind to a polypeptide expressed on the surface of an NK cell that can be used to make a cell engager provided herein (e.g., a BiKE or TriKE) include, without limitation, anti-CD16a scFvs, anti-NKG2A scFvs, anti-NKG2D scFvs, anti-NKp30 scFvs (see, e.g., BioLegend Catalog #325207), anti-NKp44 scFvs, anti-NKp46 scFvs, anti- CD16a VH domains, anti-NKG2A VH domains, anti-NKG2D VH domains, anti-NKp30 VH domains, anti-NKp44 VH domains, and anti-NKp46 VH domains.
  • amino acid sequences that can be used as antigen binding domains having the ability to bind to a polypeptide expressed on the surface of an NK cell (e.g., CD16, NKG2A, NKG2D, or NKp46) are described in McCall et al. (Mol. Immunol., 36(7):433- 445 (1999); see, e.g., anti-CD16 scFv sequences); International Patent Application Publication No. PCT/US2017/048721 (see, e.g., the CDRs and sequence listing for anti- CD16a binding domains); U.S.
  • Patent Application Publication No.2011/0052606 see, e.g., the CDRs and the sequence listing for anti-NKG2A antibodies such as Z199
  • U.S. Patent Application Publication No.2011/0150870 see, e.g., the CDRs and sequence listing for anti-NKG2D antibodies
  • U.S. Patent Application Publication No. 2018/0369373 see, e.g., the CDRs and sequence listing for anti-NKp46 antibodies
  • U.S. Patent Application Publication No.2017/0368169 see, e.g., the CDRs and sequence listing for anti-NKp46 antibodies.
  • a cell engager provided herein can be designed to include an antigen binding domain (e.g., a scFv or VH) that comprises, consists essentially of, or consists of one or more of the amino acid sequences set forth in Example 29.
  • an antigen binding domain e.g., a scFv or VH
  • a cell engager provided herein can be designed to include an antigen binding domain (e.g., a scFv or VH) that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 29 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof, provided that the antigen binding domain has the ability to bind to a polypeptide expressed on the surface of an NK cell.
  • an antigen binding domain e.g., a scFv or VH
  • a cell engager provided herein can be designed to include an antigen binding domain (e.g., a scFv or VH) that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 29 with two or less, three or less, four or less, five or less, six or less, seven or less, eight or less, nine or less, or ten or less amino acid deletions, additions, substitutions, or combinations thereof, provided that the antigen binding domain has the ability to bind to a polypeptide expressed on the surface of an NK cell.
  • a cell engager provided herein can be designed to include a linker located between each antigen binding domain.
  • Any appropriate linker can be used to design a cell engager provided herein.
  • Examples of linkers that can be used to make a cell engager described herein include, without limitation, the linker sequences set forth in Example 20.
  • a cell engager provided herein can be designed to include a linker of any appropriate length.
  • a cell engager provided herein can be designed to include a linker that is from about 3 to about 100 (e.g., from about 3 to about 90, from about 3 to about 80, from about 3 to about 70, from about 3 to about 60, from about 3 to about 50, from about 3 to about 40, from about 3 to about 30, from about 3 to about 20, from about 3 to about 15, from about 5 to about 100, from about 10 to about 100, from about 20 to about 100, from about 30 to about 100, from about 40 to about 100, from about 50 to about 100, from about 60 to about 100, from about 70 to about 100, from about 10 to about 50, from about 10 to about 40, from about 10 to about 30, from about 10 to about 20, or from about 12 to about 17) amino acid residues in length.
  • a linker that is from about 3 to about 100 (e.g., from about 3 to about 90, from about 3 to about 80, from about 3 to about 70, from about 3 to about 60, from about 3 to about 50, from about 3 to about 40, from about 3 to
  • a cell engager provided herein can be designed to include a Attorney Docket No.48881-0047WO1 / 05958 GGGGSGGGGSGGGGS (SEQ ID NO:139) linker.
  • a hinge of a CAR described herein can be used as a linker to make a cell engager described herein.
  • any one of the sequences set forth in Example 23 can be used as a linker of a cell engager described herein.
  • a cell engager provided herein can be designed to include a linker that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 20 or Example 23.
  • a cell engager provided herein can be designed to include a linker that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 20 or Example 23 with one, two, three, four, five, six, seven, eight, nine, or ten amino acid deletions, additions, substitutions, or combinations thereof.
  • a cell engager provided herein can be designed to include a linker that comprises, consists essentially of, or consists of one of the amino acid sequences set forth in Example 20 or Example 23 with two or less, three or less, four or less, five or less, six or less, seven or less, eight or less, nine or less, or ten or less amino acid deletions, additions, substitutions, or combinations thereof.
  • a cell engager e.g., a BiTE targeting a follistatin polypeptide
  • a cell engager e.g., a BiTE
  • a linker such as a linker/hinge set forth in Example 20 or Example 23 (e.g., SEQ ID NO:139 or 159), followed by an antigen binding domain having the ability to bind to a polypeptide expressed on the surface of a T cell (e.g., an anti-human CD3 scFv) (see, e.g., Example 30).
  • a cell engager e.g., a BiTE targeting a follistatin polypeptide
  • a cell engager e.g., a BiTE
  • a linker such as a linker/hinge set forth in Example 20 or Example 23 (e.g., SEQ ID NO:139 or 159), followed by an antigen binding domain having the ability to bind to a polypeptide expressed on the surface of a T cell (e.g., an anti-human CD3 scFv) (see, e.g., Example 30).
  • a cell engager e.g., a BiTE targeting a follistatin polypeptide
  • a cell engager e.g., a BiTE
  • a linker such as a linker/hinge set forth in Example 20 or Attorney Docket No.48881-0047WO1 / 05958
  • Example 23 e.g., SEQ ID NO:139 or 159
  • an antigen binding domain having the ability to bind to a polypeptide expressed on the surface of a T cell (e.g., an anti-human CD3 scFv) (see, e.g., Example 31).
  • a cell engager e.g., a BiTE targeting a follistatin polypeptide
  • a cell engager e.g., a BiTE
  • a linker such as a linker/hinge set forth in Example 20 or Example 23 (e.g., SEQ ID NO:139 or 159), followed by an antigen binding domain having the ability to bind to a polypeptide expressed on the surface of a T cell (e.g., an anti-human CD3 scFv) (see, e.g., Example 31).
  • a cell engager e.g., a BiKE or a TriKE targeting a follistatin polypeptide
  • a cell engager e.g., a BiKE or a TriKE
  • a linker such as a linker/hinge set forth in Example 20 or Example 23 (e.g., SEQ ID NO:139 or 159), followed by one or more antigen binding domains having the ability to bind to a polypeptide expressed on the surface of an NK cell (e.g., an anti-human CD16a scFv for a BiKE or an anti-human CD16a scFv and an anti-human NKG2A scFv for a TriKE).
  • a cell engager e.g., a BiKE or a TriKE targeting a follistatin polypeptide
  • a cell engager e.g., a BiKE or a TriKE
  • a linker such as a hinge/linker set forth in Example 20 or Example 23 (e.g., SEQ ID NO:139 or 159), followed by one or more antigen binding domains having the ability to bind to a polypeptide expressed on the surface of an NK cell (e.g., an anti- human CD16a scFv for a BiKE or an anti-human CD16a scFv and an anti-human NKG2A scFv for a TriKE).
  • a cell engager e.g., a BiKE or a TriKE targeting a follistatin polypeptide
  • a cell engager e.g., a BiKE or a TriKE
  • a linker such as a linker/hinge set forth in Example 20 or Example 23 (e.g., SEQ ID NO:139 or 159), followed by one or more antigen binding domains having the ability to bind to a polypeptide expressed on the surface of an NK cell (e.g., an anti-human CD16a scFv for a BiKE or an anti-human CD16a scFv and an anti-human NKG2A scFv for a TriKE).
  • a cell engager e.g., a BiKE or a TriKE
  • a cell engager targeting a follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:1 (or a variant of SEQ ID NO:1 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:2 (or a variant of SEQ ID NO:2 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:3 (or a variant of SEQ ID NO:3 with one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 3.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:1 (or a variant of SEQ ID NO:1 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:2 (or a variant of SEQ ID NO:2 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:3 (or a variant
  • such a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:4 (or a Attorney Docket No.48881-0047WO1 / 05958 variant of SEQ ID NO:4 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:5 (or a variant of SEQ ID NO:5 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:6 (or a variant of SEQ ID NO:6 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 3 can be designed to include framework regions as set forth in Example 3 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain e.g., a VH domain
  • an antibody domain can be designed to include the three CDRs set forth in Example 3 and the framework regions set forth in Example 3 except that framework region 1 having the amino acid set forth in SEQ ID NO:4 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:12, a framework region 1 having the amino acid set forth in SEQ ID NO:20, or a framework region 1 having the amino acid set forth in SEQ ID NO:28.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 3, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 Attorney Docket No.48881-0047WO1 / 05958 percent identity to the amino acid sequence set forth in SEQ ID NO:8.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include (a) a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:8.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:8, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:1, 2, and 3.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:8, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:1, 2, and 3.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:8 or the amino acid set forth in SEQ ID NO:8 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:1, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:2, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:3.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:1” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:1, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:1, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:1, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:1 include, without limitation, those set forth in Table 1. Table 1. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:1.
  • Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 G GSISSGYY 259 et forth in SEQ ID NO:2” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:2, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:2, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:2, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.
  • CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:3
  • a “CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:3” is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:3, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:3, and/or that has zero, one, two, three, four, or five amino Attorney Docket No.48881-0047WO1 / 05958 acid residues directly following SEQ ID NO:3, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a folli
  • binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide can include a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:9 (or a variant of SEQ ID NO:9 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:10 (or a variant of SEQ ID NO:10 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID
  • follistatin polypeptide e.g., a human follistatin polypeptide
  • VH domain set forth in Example 4.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • such a binder can include a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:12 (or a variant of SEQ ID NO:12 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:13 (or a variant of SEQ ID NO:13 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:14 (or a variant of SEQ ID NO:14 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4 having the amino acid sequence set forth in SEQ ID NO:15 (or a variant
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 4 can be designed to include framework regions as set forth in Example 4 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain e.g., a VH domain
  • a VH domain can be designed to include the three CDRs set forth in Example 4 and the framework regions set forth in Example 4 except that framework region 1 having the amino acid set forth in SEQ ID Attorney Docket No.48881-0047WO1 / 05958 NO:12 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:20, or a framework region 1 having the amino acid set forth in SEQ ID NO:28.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 4, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:16.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include (a) a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:16.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:16, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:9, 10, and 11.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in Attorney Docket No.48881-0047WO1 / 05958 SEQ ID NO:16, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs: 9, 10, and 11.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:16 or the amino acid set forth in SEQ ID NO:16 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:9, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:10, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:11.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:9” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:9, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:9, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:9, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists Attorney Docket No.48881-0047WO1 / 05958 essentially of the amino acid sequence set forth in SEQ ID NO:9 include, without limitation, those set forth in Table 4. Table 4. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:9.
  • a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:10 is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:10, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:10, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:10, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:10 include, without limitation, those set forth in Table 5. Table 5. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:10.
  • Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 INPNSGGT 290 INPSGGST 291 s use ere n, a t at cons sts essent a y o t e amno ac sequence set forth in SEQ ID NO:11” is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:11, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:11, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:11, provided that the binder (e.g., an antibody, antigen binding fragment
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:11 include, without limitation, those set forth in Table 6. Table 6. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:11. Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 ARIGGPIVGALTVPWMVV 305 AKDGALTVPWMEPRVV 306 , .
  • follistatin polypeptide e.g., a human follistatin polypeptide
  • a follistatin polypeptide can include a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:17 (or a variant of SEQ ID NO:17 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:18 (or a variant of SEQ ID NO:18 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:19 (or a variant of SEQ ID NO:19 with one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 5.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:17 (or a variant of SEQ ID NO:17 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:18 (or a variant of SEQ ID NO:18 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:19 (or a variant
  • such a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:20 (or a variant of SEQ ID NO:20 with one, two, three, four, five, six, seven, eight, nine, ten, or Attorney Docket No.48881-0047WO1 / 05958 more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:21 (or a variant of SEQ ID NO:21 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:22 (or a variant of SEQ ID NO:22 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 5 can be designed to include framework regions as set forth in Example 5 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain e.g., a VH domain
  • an antibody domain can be designed to include the three CDRs set forth in Example 5 and the framework regions set forth in Figure 4 except that framework region 1 having the amino acid set forth in SEQ ID NO:20 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:12, or a framework region 1 having the amino acid set forth in SEQ ID NO:28.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 5, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:24.
  • a Attorney Docket No.48881-0047WO1 / 05958 binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:24.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:24, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:17, 18, and 19.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:24, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:17, 18, and 19.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:24 or the amino acid set forth in SEQ ID NO:24 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:17, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:18, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:19.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:17” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:17, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:17, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:17, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:17 include, without limitation, those set forth in Table 7. Table 7. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:17.
  • Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 GGSISSGYY 319 et forth in SEQ ID NO:18” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:18, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:18, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:18, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:18 include, without limitation, those set forth in Table 8. Table 8. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:18.
  • a “CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:19” is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:19, that has zero, one, two, three, four, or five amino acid residues Attorney Docket No.48881-0047WO1 / 05958 directly preceding SEQ ID NO:19, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:19, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:19 include, without limitation, those set forth in Table 9. Table 9. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:19.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:25 (or a variant of SEQ ID NO:25 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:26 (or a variant of SEQ ID NO:26 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:27 (or a variant of SEQ ID NO:27 with Attorney Docket No.48881-0047WO1 / 05958 one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 6.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • such a binder can include a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:28 (or a variant of SEQ ID NO:28 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:29 (or a variant of SEQ ID NO:29 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:30 (or a variant of SEQ ID NO:30 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4 having the amino acid sequence set forth in SEQ ID NO:31 (or a variant
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 6 can be designed to include framework regions as set forth in Example 6 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain (e.g., a VH domain) can be designed to include the three CDRs set forth in Example 6 and the framework regions set forth in Attorney Docket No.48881-0047WO1 / 05958 Example 6 except that framework region 1 having the amino acid set forth in SEQ ID NO:28 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:12, or a framework region 1 having the amino acid set forth in SEQ ID NO:20.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 6, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:32.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:32, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:25, 26, and 27.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in Attorney Docket No.48881-0047WO1 / 05958 SEQ ID NO:32, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:25, 26, and 27.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:32 or the amino acid set forth in SEQ ID NO:32 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:25, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:26, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:27.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:25” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:25, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:25, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:25, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists Attorney Docket No.48881-0047WO1 / 05958 essentially of the amino acid sequence set forth in SEQ ID NO:25 include, without limitation, those set forth in Table 10.
  • Table 10 Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:25.
  • a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:26” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:26, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:26, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:26, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:26 include, without limitation, those set forth in Table 11. Table 11. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:26.
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:27 include, without limitation, those set forth in Table 12. Table 12. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:27. Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 ARIGLRSSGSRESHPI 365 AKDGLRSSGSRESHPI 366 , .
  • follistatin polypeptide e.g., a human follistatin polypeptide
  • a follistatin polypeptide can include a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:33 (or a variant of SEQ ID NO:33 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:34 (or a variant of SEQ ID NO:34 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:35 (or a variant of SEQ ID NO:35 with one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 7.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:33 (or a variant of SEQ ID NO:33 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:34 (or a variant of SEQ ID NO:34 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:35 (or a variant
  • such a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:36 (or a variant of SEQ ID NO:36 with one, two, three, four, five, six, seven, eight, nine, ten, or Attorney Docket No.48881-0047WO1 / 05958 more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:37 (or a variant of SEQ ID NO:37 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:38 (or a variant of SEQ ID NO:38 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 7 can be designed to include framework regions as set forth in Example 7 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain e.g., a VH domain
  • an antibody domain can be designed to include the three CDRs set forth in Example 7 and the framework regions set forth in Example 7 except that framework region 1 having the amino acid set forth in SEQ ID NO:36 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:12, a framework region 1 having the amino acid set forth in SEQ ID NO:20, or a framework region 1 having the amino acid set forth in SEQ ID NO:28.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 7, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:40.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:40.
  • a Attorney Docket No.48881-0047WO1 / 05958 binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:40.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:40, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:33, 34, and 35.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:40, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs: 33, 34, and 35.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:40 or the amino acid set forth in SEQ ID NO:40 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:33, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:34, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:35.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:33” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:33, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:33, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:33, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:33 include, without limitation, those set forth in Table 13. Table 13. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:33 Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 G GSISSGYY 238 et forth in SEQ ID NO:34” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:34, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:34, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:34, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:34 include, without limitation, those set forth in Table 14. Table 14. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:34.
  • a “CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:35” is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:35, that has zero, one, two, three, four, or five amino acid residues Attorney Docket No.48881-0047WO1 / 05958 directly preceding SEQ ID NO:35, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:35, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:35 include, without limitation, those set forth in Table 15. Table 15. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:35.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:41 (or a variant of SEQ ID NO:41 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:42 (or a variant of SEQ ID NO:42 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:43 (or a variant of SEQ ID NO:43 with Attorney Docket No.48881-0047WO1 / 05958 one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 8.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:41 (or a variant of SEQ ID NO:41 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:42 (or a variant of SEQ ID NO:42 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:43 (or a variant
  • such a binder can include a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:44 (or a variant of SEQ ID NO:44 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:45 (or a variant of SEQ ID NO:45 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:46 (or a variant of SEQ ID NO:46 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4 having the amino acid sequence set forth in SEQ ID NO:47 (or a variant
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 8 can be designed to include framework regions as set forth in Example 8 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain (e.g., a VH domain) can be designed to include the three CDRs set forth in Example 8 and the framework regions set forth in Attorney Docket No.48881-0047WO1 / 05958 Example 8 except that framework region 1 having the amino acid set forth in SEQ ID NO:44 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:20, or a framework region 1 having the amino acid set forth in SEQ ID NO:28.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 8, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:48.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:48.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:48, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:41, 42, and 43.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in Attorney Docket No.48881-0047WO1 / 05958 SEQ ID NO:48, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:41, 42, and 43.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:48 or the amino acid set forth in SEQ ID NO:48 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:41, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:42, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:43.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:41” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:41, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:41, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:41, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists Attorney Docket No.48881-0047WO1 / 05958 essentially of the amino acid sequence set forth in SEQ ID NO:41 include, without limitation, those set forth in Table 16. Table 16. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:41.
  • a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:42 is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:42, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:42, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:42, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:42 include, without limitation, those set forth in Table 17. Table 17. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:42.
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:43 include, without limitation, those set forth in Table 18. Table 18. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:43. Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 ARIGAPREMATI 404 AKDGAPREMATI 405 , .
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a follistatin polypeptide can include a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:49 (or a variant of SEQ ID NO:49 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:50 (or a variant of SEQ ID NO:50 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:51 (or a variant of SEQ ID NO:51 with one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 9.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:49 (or a variant of SEQ ID NO:49 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:50 (or a variant of SEQ ID NO:50 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:51 (or a variant
  • such a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:52 (or a variant of SEQ ID NO:52 with one, two, three, four, five, six, seven, eight, nine, ten, or Attorney Docket No.48881-0047WO1 / 05958 more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:53 (or a variant of SEQ ID NO:53 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:54 (or a variant of SEQ ID NO:54 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 9 can be designed to include framework regions as set forth in Example 9 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain e.g., a VH domain
  • an antibody domain can be designed to include the three CDRs set forth in Example 9 and the framework regions set forth in Example 9 except that framework region 1 having the amino acid set forth in SEQ ID NO:52 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:12, or a framework region 1 having the amino acid set forth in SEQ ID NO:28.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 9, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:56.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:56.
  • a Attorney Docket No.48881-0047WO1 / 05958 binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:56.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:56, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:49, 50, and 51.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:56, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:49, 50, and 51.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:56 or the amino acid set forth in SEQ ID NO:56 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:49, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:50, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:51.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:49” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:49, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:49, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:49, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:49 include, without limitation, those set forth in Table 19. Table 19. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:49.
  • Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 GGSISSGYY 418 et forth in SEQ ID NO:50” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:50, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:50, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:50, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:50 include, without limitation, those set forth in Table 20. Table 20. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:50.
  • a “CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:51” is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:51, that has zero, one, two, three, four, or five amino acid residues Attorney Docket No.48881-0047WO1 / 05958 directly preceding SEQ ID NO:51, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:51, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:51 include, without limitation, those set forth in Table 21. Table 21. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:51.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:57 (or a variant of SEQ ID NO:57 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:58 (or a variant of SEQ ID NO:58 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:59 (or a variant of SEQ ID NO:59 with Attorney Docket No.48881-0047WO1 / 05958 one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 10.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:57 (or a variant of SEQ ID NO:57 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:58 (or a variant of SEQ ID NO:58 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:59 (or a variant
  • such a binder can include a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:60 (or a variant of SEQ ID NO:60 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:61 (or a variant of SEQ ID NO:61 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:62 (or a variant of SEQ ID NO:62 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4 having the amino acid sequence set forth in SEQ ID NO:63 (or a variant
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 10 can be designed to include framework regions as set forth in Example 10 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain (e.g., a VH domain) can be designed to include the three CDRs set forth in Example 10 and the framework regions Attorney Docket No.48881-0047WO1 / 05958 set forth in Example 10 except that framework region 1 having the amino acid set forth in SEQ ID NO:60 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:12, or a framework region 1 having the amino acid set forth in SEQ ID NO:20.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 10, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:64.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:64.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:64, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:57, 58, and 59.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in Attorney Docket No.48881-0047WO1 / 05958 SEQ ID NO:64, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:57, 58, and 59.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:64 or the amino acid set forth in SEQ ID NO:64 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:57, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:58, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:59.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:57” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:57, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:57, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:57, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists Attorney Docket No.48881-0047WO1 / 05958 essentially of the amino acid sequence set forth in SEQ ID NO:57 include, without limitation, those set forth in Table 22. Table 22. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:57.
  • a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:58” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:58, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:58, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:58, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:58 include, without limitation, those set forth in Table 23. Table 23. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:58.
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:59 include, without limitation, those set forth in Table 24. Table 24. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:59. Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 ARIGAFYTWNL 464 AKDGAFYTWNL 465 , .
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a follistatin polypeptide can include a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:65 (or a variant of SEQ ID NO:65 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:66 (or a variant of SEQ ID NO:66 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:67 (or a variant of SEQ ID NO:67 with one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 11.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:65 (or a variant of SEQ ID NO:65 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:66 (or a variant of SEQ ID NO:66 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:67 (or a variant
  • such a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:68 (or a variant of SEQ ID NO:68 with one, two, three, four, five, six, seven, eight, nine, ten, or Attorney Docket No.48881-0047WO1 / 05958 more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:69 (or a variant of SEQ ID NO:69 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:70 (or a variant of SEQ ID NO:70 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 11 can be designed to include framework regions as set forth in Example 11 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain e.g., a VH domain
  • an antibody domain can be designed to include the three CDRs set forth in Example 11 and the framework regions set forth in Example 11 except that framework region 1 having the amino acid set forth in SEQ ID NO:68 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:12, or a framework region 1 having the amino acid set forth in SEQ ID NO:20.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 11, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:72.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:72.
  • a Attorney Docket No.48881-0047WO1 / 05958 binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:72.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:72, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:65, 66, and 67.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:72, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:65, 66, and 67.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:72 or the amino acid set forth in SEQ ID NO:72 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:65, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:66, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:67.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:65” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:65, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:65, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:65, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:65 include, without limitation, those set forth in Table 25. Table 25. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:65.
  • Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 G GSISSGYY 478 et forth in SEQ ID NO:66” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:66, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:66, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:66, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:66 include, without limitation, those set forth in Table 26. Table 26. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:66.
  • a “CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:67” is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:67, that has zero, one, two, three, four, or five amino acid residues Attorney Docket No.48881-0047WO1 / 05958 directly preceding SEQ ID NO:67, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:67, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:67 include, without limitation, those set forth in Table 27. Table 27. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:67.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:73 (or a variant of SEQ ID NO:73 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:74 (or a variant of SEQ ID NO:74 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:75 (or a variant of SEQ ID NO:75 with Attorney Docket No.48881-0047WO1 / 05958 one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 12.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:73 (or a variant of SEQ ID NO:73 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:74 (or a variant of SEQ ID NO:74 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:75 (or a variant
  • such a binder can include a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:76 (or a variant of SEQ ID NO:76 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:77 (or a variant of SEQ ID NO:77 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:78 (or a variant of SEQ ID NO:78 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4 having the amino acid sequence set forth in SEQ ID NO:79 (or a variant
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 12 can be designed to include framework regions as set forth in Example 12 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain (e.g., a VH domain) can be designed to include the three CDRs set forth in Example 12 and the framework regions Attorney Docket No.48881-0047WO1 / 05958 set forth in Example 12 except that framework region 1 having the amino acid set forth in SEQ ID NO:76 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:20, or a framework region 1 having the amino acid set forth in SEQ ID NO:28.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 12, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:80.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:80.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:80, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:73, 74, and 75.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in Attorney Docket No.48881-0047WO1 / 05958 SEQ ID NO:80, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:73, 74, and 75.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:80 or the amino acid set forth in SEQ ID NO:80 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:73, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:74, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:75.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:73” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:73, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:73, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:73, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists Attorney Docket No.48881-0047WO1 / 05958 essentially of the amino acid sequence set forth in SEQ ID NO:73 include, without limitation, those set forth in Table 28. Table 28. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:73.
  • a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:74” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:74, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:74, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:74, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:74 include, without limitation, those set forth in Table 29. Table 29. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:74.
  • Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 INPNSGGT 509 INPSGGST 510 s use ere n, a t at cons sts essent a y o t e amno ac sequence set forth in SEQ ID NO:75” is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:75, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:75, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:75, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:75 include, without limitation, those set forth in Table 30. Table 30. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:75. Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 ARIDGYYGMMV 524 AKDGDGYYGMDV 525 , .
  • follistatin polypeptide e.g., a human follistatin polypeptide
  • a follistatin polypeptide can include a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:81 (or a variant of SEQ ID NO:81 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:82 (or a variant of SEQ ID NO:82 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:83 (or a variant of SEQ ID NO:83 with one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 13.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • such a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:84 (or a variant of SEQ ID NO:84 with one, two, three, four, five, six, seven, eight, nine, ten, or Attorney Docket No.48881-0047WO1 / 05958 more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:85 (or a variant of SEQ ID NO:85 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:86 (or a variant of SEQ ID NO:86 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 13 can be designed to include framework regions as set forth in Example 13 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain e.g., a VH domain
  • an antibody domain can be designed to include the three CDRs set forth in Example 13 and the framework regions set forth in Example 13 except that framework region 1 having the amino acid set forth in SEQ ID NO:84 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:12, or a framework region 1 having the amino acid set forth in SEQ ID NO:28.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 13, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:88.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:88.
  • a Attorney Docket No.48881-0047WO1 / 05958 binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:88.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:88, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:81, 82, and 83.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:88, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:81, 82, and 83.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:88 or the amino acid set forth in SEQ ID NO:88 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:81, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:82, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:83.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:81” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:81, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:81, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:81, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:81 include, without limitation, those set forth in Table 31. Table 31. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:81.
  • Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 GGSISSGYY 538 et forth in SEQ ID NO:82” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:82, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:82, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:82, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:82 include, without limitation, those set forth in Table 32. Table 32. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:82.
  • a “CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:83” is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:83, that has zero, one, two, three, four, or five amino acid residues Attorney Docket No.48881-0047WO1 / 05958 directly preceding SEQ ID NO:83, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:83, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:83 include, without limitation, those set forth in Table 33. Table 33. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:83.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:89 (or a variant of SEQ ID NO:89 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:90 (or a variant of SEQ ID NO:90 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:91 (or a variant of SEQ ID NO:91 with Attorney Docket No.48881-0047WO1 / 05958 one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 14.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:89 (or a variant of SEQ ID NO:89 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:90 (or a variant of SEQ ID NO:90 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:91 (or a variant
  • such a binder can include a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:92 (or a variant of SEQ ID NO:92 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:93 (or a variant of SEQ ID NO:93 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:94 (or a variant of SEQ ID NO:94 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4 having the amino acid sequence set forth in SEQ ID NO:95 (or a variant
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 14 can be designed to include framework regions as set forth in Example 14 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain (e.g., a VH domain) can be designed to include the three CDRs set forth in Example 14 and the framework regions Attorney Docket No.48881-0047WO1 / 05958 set forth in Example 14 except that framework region 1 having the amino acid set forth in SEQ ID NO:92 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:12, or a framework region 1 having the amino acid set forth in SEQ ID NO:20.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 14, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:96.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:96.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include (a) a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:96.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:96, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:89, 90, and 91.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in Attorney Docket No.48881-0047WO1 / 05958 SEQ ID NO:96, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:89, 90, and 91.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:96 or the amino acid set forth in SEQ ID NO:96 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:89, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:90, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:91.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:89” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:89, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:89, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:89, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists Attorney Docket No.48881-0047WO1 / 05958 essentially of the amino acid sequence set forth in SEQ ID NO:89 include, without limitation, those set forth in Table 34. Table 34. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:89.
  • a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:90 is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:90, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:90, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:90, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:90 include, without limitation, those set forth in Table 35. Table 35. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:90.
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:91 include, without limitation, those set forth in Table 36. Table 36. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:91. Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 ARIGRRVYDSSGYYVMV 584 AKDGRRVYDSSGYYV 585 , .
  • follistatin polypeptide e.g., a human follistatin polypeptide
  • a follistatin polypeptide can include a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:97 (or a variant of SEQ ID NO:97 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:98 (or a variant of SEQ ID NO:98 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:99 (or a variant of SEQ ID NO:99 with one or two amino acid modifications).
  • a binder having these CDRs and the ability to bind to a follistatin polypeptide includes, without limitation, the VH domain set forth in Example 15.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • having a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:97 (or a variant of SEQ ID NO:97 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:98 (or a variant of SEQ ID NO:98 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:99 (or a variant
  • such a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:100 (or a variant of SEQ ID NO:100 with one, two, three, four, five, six, seven, eight, nine, ten, or Attorney Docket No.48881-0047WO1 / 05958 more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:101 (or a variant of SEQ ID NO:101 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:102 (or a variant of SEQ ID NO:102 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 15 can be designed to include framework regions as set forth in Example 15 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain e.g., a VH domain
  • an antibody domain can be designed to include the three CDRs set forth in Example 15 and the framework regions set forth in Example 15 except that framework region 1 having the amino acid set forth in SEQ ID NO:100 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:12, or a framework region 1 having the amino acid set forth in SEQ ID NO:28.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 15, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:104.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:104.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:104.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:104, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:97, 98, and 99.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:104, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:97, 98, and 99.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:104 or the amino acid set forth in SEQ ID NO:104 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:97, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:98, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:99.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:97” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:97, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:97, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:97, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:97 include, without limitation, those set forth in Table 37. Table 37. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:97.
  • Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 GGSISSGYY 598 et forth in SEQ ID NO:98” is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:98, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:98, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:98, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:98 include, without limitation, those set forth in Table 38. Table 38. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:98.
  • a “CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:99” is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:99, that has zero, one, two, three, four, or five amino acid residues Attorney Docket No.48881-0047WO1 / 05958 directly preceding SEQ ID NO:99, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:99, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:99 include, without limitation, those set forth in Table 39. Table 39. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:99.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having a CDR1 having the amino acid sequence set forth in SEQ ID NO:105 (or a variant of SEQ ID NO:105 with one or two amino acid modifications), a CDR2 having the amino acid sequence set forth in SEQ ID NO:106 (or a variant of SEQ ID NO:106 with one or two amino acid modifications), and a CDR3 having the amino acid sequence set forth in SEQ ID NO:107 (or a variant of SEQ ID Attorney Docket No.48881-0047WO1 / 05958 NO:107 with one or two amino acid modifications).
  • such a binder can include a heavy chain variable domain that includes a framework region 1 having the amino acid sequence set forth in SEQ ID NO:108 (or a variant of SEQ ID NO:108 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 2 having the amino acid sequence set forth in SEQ ID NO:109 (or a variant of SEQ ID NO:109 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), a framework region 3 having the amino acid sequence set forth in SEQ ID NO:110 (or a variant of SEQ ID NO:110 with one, two, three, four, five, six, seven, eight, nine, ten, or more amino acid modifications), and a framework region 4 having the amino acid sequence set forth in SEQ ID NO:111 (or a variant
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder having any of the CDRs set forth in Example 16 can be designed to include framework regions as set forth in Example 16 or can be designed to include one or more framework regions from another antibody or antibody fragment.
  • an antibody domain (e.g., a VH domain) can be designed to include the three CDRs set forth in Example 16 and the framework regions Attorney Docket No.48881-0047WO1 / 05958 set forth in Example 16 except that framework region 1 having the amino acid set forth in SEQ ID NO:108 is replaced with a framework region 1 having the amino acid set forth in SEQ ID NO:4, a framework region 1 having the amino acid set forth in SEQ ID NO:12, or a framework region 1 having the amino acid set forth in SEQ ID NO:20.
  • an Fab or scFv can be designed to include (a) the three CDRs set forth in Example 16, (b) the framework regions set forth in Examples 3-16, and (c) a light chain variable domain.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:112.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence set forth in SEQ ID NO:112.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a heavy chain variable domain that includes an amino acid sequence having 100 percent identity to the amino acid sequence set forth in SEQ ID NO:112.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain that includes an amino acid sequence having at least 90 percent identity to the amino acid sequence set forth in SEQ ID NO:112, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:105, 106, and 107.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a binder can include a heavy chain variable domain that includes an amino acid sequence having at least 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 percent identity to the amino acid sequence Attorney Docket No.48881-0047WO1 / 05958 set forth in SEQ ID NO:112, provided that the heavy chain variable domain includes the amino acid sequences set forth in SEQ ID NOs:105, 106, and 107.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain having the amino acid sequence set forth in SEQ ID NO:112 or the amino acid set forth in SEQ ID NO:112 with one, two, three, four, five, six, seven, eight, nine, or 10 amino acid modifications (e.g., amino acid substitutions, amino acid deletions, and/or amino acid additions).
  • antibody domain e.g., a VH domain
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a heavy chain variable domain comprising (i) a CDR1 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:105, (ii) a CDR2 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:106, and (iii) a CDR3 that comprises, consists essentially of, or consists of the amino acid sequence set forth in SEQ ID NO:107.
  • a “CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:105” is a CDR1 that has zero, one, or two amino acid substitutions within SEQ ID NO:105, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:105, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:105, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a Attorney Docket No.48881-0047WO1 / 05958 follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • Examples of a CDR1 that consists essentially of the amino acid sequence set forth in SEQ ID NO:105 include, without limitation, those set forth in Table 40. Table 40. Exemplary CDR1s that consist essentially of the amino acid sequence set forth in SEQ ID NO:105.
  • a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:106 is a CDR2 that has zero, one, or two amino acid substitutions within SEQ ID NO:106, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:106, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:106, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR2 that consists essentially of the amino acid sequence set forth in SEQ ID NO:106 include, without limitation, those set forth in Table 41. Table 41. Exemplary CDR2s that consist essentially of the amino acid sequence set forth in SEQ ID NO:106.
  • a C 3 t at cons sts essent a y o t e amno acd sequence set forth in SEQ ID NO:107 is a CDR3 that has zero, one, or two amino acid substitutions within SEQ ID NO:107, that has zero, one, two, three, four, or five amino acid residues directly preceding SEQ ID NO:107, and/or that has zero, one, two, three, four, or five amino acid residues directly following SEQ ID NO:107, provided that the binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) maintains its basic ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Examples of a CDR3 that consists essentially of the amino acid sequence set forth in SEQ ID NO:107 include, without limitation, those set forth in Table 42. Table 42. Exemplary CDR3s that consist essentially of the amino acid sequence set forth in SEQ ID NO:107. Sequence SEQ ID NO: Attorney Docket No.48881-0047WO1 / 05958 ARRGLFSGVPL 643 ARILFSGVPLL 644 . ., variable domain and a light chain variable domain, the two regions can be directly connected or can be connected using any appropriate linker sequence. For example, a heavy chain variable domain can be connected to a light chain variable domain via a linker sequence.
  • the amino acid sequences described herein can include amino acid modifications (e.g., the articulated number of amino acid modifications). Such amino acid modifications can include, without limitation, amino acid substitutions, amino acid deletions, amino acid additions, and combinations.
  • an amino acid modification can be made to improve the binding and/or contact with an antigen and/or to improve a functional activity of a binder (e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC) provided herein.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, a CAR, a cell engager, and/or an ADC
  • an amino acid substitution within an articulated sequence identifier can be a conservative amino acid substitution.
  • conservative amino acid substitutions can be made by substituting one amino acid residue for another amino acid residue having a similar side chain.
  • Families of amino acid residues having similar side chains can include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta- Attorney Docket No.48881-0047WO1 / 05958 branched side chains (e.g., threonine, valine, isoleucine),
  • an amino acid substitution within an articulated sequence identifier can be a non-conservative amino acid substitution.
  • Non-conservative amino acid substitutions can be made by substituting one amino acid residue for another amino acid residue having a dissimilar side chain.
  • Examples of non-conservative substitutions include, without limitation, substituting (a) a hydrophilic residue (e.g., serine or threonine) for a hydrophobic residue (e.g., leucine, isoleucine, phenylalanine, valine, or alanine); (b) a cysteine or proline for any other residue; (c) a residue having a basic side chain (e.g., lysine, arginine, or histidine) for a residue having an acidic side chain (e.g., aspartic acid or glutamic acid); and (d) a residue having a bulky side chain (e.g., phenylalanine) for glycine or other residue having a small
  • the percent sequence identity between a particular amino acid or nucleic acid sequence and an amino acid or nucleic acid sequence referenced by a particular sequence identification number is determined as follows. First, an amino acid or nucleic acid sequence is compared to the sequence set forth in a particular sequence identification number using the BLAST 2 Sequences (Bl2seq) program from the stand-alone version of BLASTZ containing BLASTN version 2.0.14 and BLASTP version 2.0.14. This stand- alone version of BLASTZ can be obtained from Fish & Richardson’s web site (e.g., www.fr.com/blast/) or the U.S. government’s National Center for Biotechnology Information web site (www.ncbi.nlm.nih.gov).
  • Bl2seq performs a comparison between two sequences using either the BLASTN or BLASTP algorithm.
  • BLASTN is used to compare nucleic acid sequences
  • BLASTP is used to compare amino acid sequences.
  • the options are set as follows: -i is set to a file containing the first nucleic acid sequence to be compared (e.g., C: ⁇ seq1.txt); -j is set to a file containing the second nucleic acid sequence to be compared (e.g., C: ⁇ seq2.txt); -p is set to blastn; -o is set to any desired file name (e.g., C: ⁇ output.txt); -q is set to -1; -r is set to 2; and all other options are left at their default setting.
  • -i is set to a file containing the first nucleic acid sequence to be compared (e.g., C: ⁇ seq1.txt)
  • -j is set to a file containing the second nucleic acid sequence to be compared (e.g., C: ⁇ seq2.txt)
  • -p is set to blastn
  • -o is set to any desired file name (e
  • the following command can be used to generate an output Attorney Docket No.48881-0047WO1 / 05958 file containing a comparison between two sequences: C: ⁇ Bl2seq -i c: ⁇ seq1.txt -j c: ⁇ seq2.txt -p blastn -o c: ⁇ output.txt -q -1 -r 2.
  • Bl2seq are set as follows: -i is set to a file containing the first amino acid sequence to be compared (e.g., C: ⁇ seq1.txt); -j is set to a file containing the second amino acid sequence to be compared (e.g., C: ⁇ seq2.txt); -p is set to blastp; -o is set to any desired file name (e.g., C: ⁇ output.txt); and all other options are left at their default setting.
  • -i is set to a file containing the first amino acid sequence to be compared (e.g., C: ⁇ seq1.txt)
  • -j is set to a file containing the second amino acid sequence to be compared (e.g., C: ⁇ seq2.txt)
  • -p is set to blastp
  • -o is set to any desired file name (e.g., C: ⁇ output.txt); and all other options are left at
  • the following command can be used to generate an output file containing a comparison between two amino acid sequences: C: ⁇ Bl2seq -i c: ⁇ seq1.txt -j c: ⁇ seq2.txt -p blastp -o c: ⁇ output.txt. If the two compared sequences share homology, then the designated output file will present those regions of homology as aligned sequences. If the two compared sequences do not share homology, then the designated output file will not present aligned sequences. Once aligned, the number of matches is determined by counting the number of positions where an identical nucleotide or amino acid residue is presented in both sequences.
  • a matched position refers to a position in which an identical nucleotide or amino acid residue occurs at the same position in aligned sequences.
  • the percent sequence identity is determined by dividing the number of matches by the length of the sequence set forth in the identified sequence (e.g., SEQ ID NO:8, SEQ ID NO:16, or SEQ ID NO:24), followed by multiplying the resulting value by 100.
  • Methods for generating an amino acid sequence variant can include site-specific mutagenesis or random mutagenesis (e.g., by PCR) of a nucleic acid encoding the antibody or fragment thereof. See, for example, Zoller, Curr. Opin. Biotechnol.3: 348-354 (1992).
  • Both naturally occurring and non-naturally Attorney Docket No.48881-0047WO1 / 05958 occurring amino acids can be used to generate an amino acid sequence variant provided herein.
  • a representative number of binders e.g., antibodies, antigen binding fragments, and/or antibody domains
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Table 43 Representative number of binders.
  • binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, cell engagers, and/or ADCs
  • the binders can be produced using any appropriate method.
  • the binders e.g., antibodies, antigen binding fragments, antibody domains, CARs, and/or cell engagers
  • the binders can be produced in recombinant host cells.
  • a nucleic acid encoding a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • Example 17 is a sequence listing of nucleic acid sequences Attorney Docket No.48881-0047WO1 / 05958 encoding exemplary binders (e.g., antibodies, antigen binding fragments, and/or antibody domains) described herein.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • prokaryotic hosts such as E.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • yeast e.g., Pichia pastoris, Saccharomyces cerevisiae, Hansenula polymorpha, Schizosaccharomyces pombe, Schwanniomyces occidentalis, Kluyveromyces lactis, or Yarrowia lipolytica
  • filamentous fungi of the genera Trichoderma e.g., T.
  • an antigen binding fragment or antibody domain provided herein can be produced by proteolytic digestion of an intact antibody.
  • an antigen binding fragment can be obtained by treating an antibody with an enzyme such as papain or pepsin.
  • Papain digestion of whole antibodies can be used to produce F(ab) 2 or Fab fragments, while pepsin digestion of whole antibodies can be used to produce F(ab’) 2 or Fab’ fragments.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • substantially pure refers to the binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC) as being substantially free of other polypeptides, lipids, carbohydrates, and nucleic acid with which it is naturally associated.
  • a substantially pure binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • any binder e.g., an antibody, antigen Attorney Docket No.48881-0047WO1 / 05958 binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a substantially pure binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a substantially pure binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • bispecific binders e.g., bispecific antibodies, bispecific antigen binding fragments, and/or bispecific antibody domains
  • a bispecific binder provided herein can be designed to bind to two different epitopes of the same follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a bispecific binder provided herein can bind to a follistatin polypeptide (e.g., a human follistatin polypeptide) and to an epitope on a different polypeptide (e.g., a CD3 polypeptide).
  • Bispecific binders can be produced by chemically conjugating two different binders (e.g., antibodies, antigen binding fragments, and/or antibody domains) together.
  • Bispecific binders also can be produced by fusing two antibody-producing cells, e.g., hybridomas, to make a hybrid cell line that produces two different heavy and two different light chains within the same cell, which can result in, for example, bispecific IgG molecules.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • a binder can be fused or conjugated (e.g., covalently or non-covalently attached) to another polypeptide or other moiety to provide a fusion protein or conjugate.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • a polymer e.g., polyethylene glycol (PEG), polyethylenimine (PEI) modified with PEG (PEI-PEG), and/or polyglutamic acid (PGA) (N-(2-Hydroxypropyl) methacrylamide (HPMA) copolymers
  • HPMA polyglutamic acid copolymers
  • hyaluronic acid e.g., a fluorescent substance, a luminescent substance, a hapten, an enzyme, a metal chelate, a drug, a radioisotope, and/or a cytotoxic agent.
  • any appropriate method can be used to conjugate (e.g., covalently or non-covalently attach) another polypeptide or other moiety to a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or Attorney Docket No.48881-0047WO1 / 05958 cell engager) provided herein.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or Attorney Docket No.48881-0047WO1 / 05958 cell engager
  • another polypeptide or other moiety can be conjugated to a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager) provided herein using the methods described in U.S. Patent No.8,021,661.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • substantially non-antigenic polymers examples include, without limitation, polyalkylene oxides and polyethylene oxides.
  • a polymer used herein can have any appropriate molecule weight.
  • a polymer having an average molecular weight from about 200 Daltons to about 35,000 Daltons (e.g., from about 1,000 to about 15,000 Daltons or from about 2,000 to about 12,500 Daltons) can be used.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • ADC an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • water soluble polymers examples include, without limitation, hydrophilic polyvinyl polymers, polyvinylalcohol, polyvinylpyrrolidone, polyalkylene oxide homopolymers, polyethylene glycol (PEG), polypropylene glycols, polyoxyethylenated polyols, and copolymers thereof and/or block copolymers thereof provided that the water solubility of the copolymer or block copolymers is maintained.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a binder can be attached (e.g., covalently or non-covalently attached) to one or more polyoxyalkylenes (e.g., polyoxyethylene, polyoxypropylene, or block copolymers of polyoxyethylene and polyoxypropylene), polymethacrylates, carbomers, branched or unbranched polysaccharides, or combinations thereof.
  • a binder e.g., an antibody, Attorney Docket No.48881-0047WO1 / 05958 antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • ADC refers to a conjugate that includes (a) an antigen binding domain and (b) at least one drug covalently linked directly or indirectly to that antigen binding domain.
  • an ADC described herein can include (a) an antigen binding domain having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide) and (b) at least one drug covalently linked directly or indirectly to that antigen binding domain.
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • Any appropriate binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • having the ability to bind to a follistatin polypeptide e.g., a human follistatin polypeptide
  • any of the binders set forth in Table 43 can be used to make an ADC having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • drugs that can be used to make an ADC described herein include, without limitation, auristatins (e.g., monomethyl auristatin E (MMAE)), mertansine (DM-1), and pyrrolobenzodiazepine (PBD) dimers.
  • auristatins e.g., monomethyl auristatin E (MMAE)
  • DM-1 mertansine
  • PBD pyrrolobenzodiazepine
  • Any appropriate ADC linker can be used to covalently attach one or more drugs to an antigen binding domain having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide) to form an ADC provided herein.
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • cleavable or non-cleavable ADC linkers can be used to covalently attach one or more drugs to an antigen binding domain having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide) to form an ADC provided herein.
  • ADC linkers can be used to covalently attach one or more drugs to an antigen binding domain having the ability to bind to a follistatin polypeptide (e.g., a human follistatin polypeptide) to form an ADC provided herein include, without limitation, ADC disulfide linkers, ADC hydrazone linkers, ADC peptide linkers, ADC thioether linkers, and ADC PEG-containing linkers.
  • This document also provides nucleic acid molecules (e.g., isolated nucleic acid molecules) having a nucleic acid sequence encoding at least part of a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager) provided herein.
  • an isolated nucleic acid molecule provided herein can Attorney Docket No.48881-0047WO1 / 05958 include a nucleic acid sequence encoding a VH domain such as a VH domain as set forth in any one of Examples 3-16.
  • an isolated nucleic acid molecule provided herein can include a nucleic acid sequence encoding a CAR or cell engager (e.g., a BiTE, BiKE, or TriKE) described herein.
  • a nucleic acid provided herein e.g., an isolated nucleic acid molecule
  • can be single stranded or double stranded nucleic acid of any appropriate type e.g., DNA, RNA, or DNA/RNA hybrids).
  • vectors e.g., plasmid vectors or viral vectors
  • plasmid vectors or viral vectors containing one or more nucleic acids provided herein.
  • An example of a plasmid vector that can be designed to include one or more nucleic acids having a nucleic acid sequence encoding at least part of a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager) provided herein includes, without limitation, phagemids.
  • viral vectors that can be designed to include one or more nucleic acids having a nucleic acid sequence encoding at least part of a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager) provided herein include, without limitation, retroviral vectors, parvovirus-based vectors (e.g., adenoviral-based vectors and adeno-associated virus (AAV)-based vectors), lentiviral vectors (e.g., herpes simplex (HSV)-based vectors), poxviral vectors (e.g., vaccinia virus-based vectors and fowlpox virus-based vectors), and hybrid or chimeric viral vectors.
  • retroviral vectors e.g., parvovirus-based vectors (e.g., adenoviral-based vectors and adeno-associated virus (AAV)-based vectors), lentiviral vectors (e.g., herpes simplex
  • a viral vector having an adenoviral backbone with lentiviral components such as those described elsewhere (Zheng et al., Nat. Biotech., 18(2): 176-80 (2000); WO 98/22143; WO 98/46778; and WO 00/17376) or viral vectors having an adenoviral backbone with AAV components such as those described elsewhere (Fisher et al., Hum. Gene Ther., 7:2079-2087 (1996)) can be designed to include one or more nucleic acids having a nucleic acid sequence encoding at least part of a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager) provided herein.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • a vector e.g., a plasmid vector or a viral vector
  • a vector can include a nucleic acid sequence encoding scFv or antibody domain (e.g., a VH domain) provided herein.
  • a vector e.g., a plasmid vector or a viral vector
  • a nucleic acid sequence encoding CAR provided herein.
  • a vector e.g., a plasmid vector or a viral vector
  • a nucleic acid sequence encoding cell engager provided herein.
  • a vector provided herein can include any appropriate promoter and other regulatory sequence (e.g., transcription and translation initiation and termination codons) operably linked the nucleic acid sequence encoding at least part of a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager) provided herein.
  • a promoter used to drive expression can be a constitutive promotor or a regulatable promotor. Examples of regulatable promoters that can be used as described herein include, without limitation, inducible promotors, repressible promotors, and tissue-specific promoters.
  • viral promotors examples include, without limitation, adenoviral promotors, vaccinia virus promotors, CMV promotors (e.g., immediate early CMV promotors), and AAV promoters. Any appropriate method can be used to make a nucleic acid molecule (or vector such as a plasmid vector or viral vector) having a nucleic acid sequence encoding at least part of a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager) provided herein.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • nucleic acid molecule or vector such as a plasmid vector or viral vector
  • a nucleic acid sequence encoding at least part of a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager) provided herein as described elsewhere (see, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory, NY (1989); and Ausubel et al., Current Protocols in Molecular Biology, Green Publishing Associates and John Wiley & Sons, New York, N.Y. (1994)).
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • This document also provides host cells that include a nucleic acid provided herein (e.g., a nucleic acid having a nucleic acid sequence encoding at least part of a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager) provided herein).
  • Host cells that can be designed to include one or more nucleic acids provided herein can be prokaryotic cells or eukaryotic cells. Examples of prokaryotic cells that can be designed to include a nucleic acid provided herein include, without Attorney Docket No.48881-0047WO1 / 05958 limitation, E.
  • coli e.g., Tb-1, TG-1, DH5?, XL-Blue MRF (Stratagene), SA2821, or Y1090 cells
  • Bacillus subtilis Bacillus subtilis, Salmonella typhimurium, Serratia marcescens, or Pseudomonas (e.g., P. aerugenosa) cells.
  • eukaryotic cells that can be designed to include a nucleic acid provided herein include, without limitation, insect cells (e.g., Sf9 or Ea4 cells), yeast cells (e.g., S. cerevisiae cells), and mammalian cells (e.g., mouse, rat, hamster, monkey, or human cells).
  • VERO cells can be designed to include a nucleic acid provided herein.
  • Any appropriate method can be used to introduce one or more nucleic acids provided herein (e.g., a vector such as a plasmid vector or viral vector having a nucleic acid sequence encoding at least part of a binder provided herein) into a host cell.
  • calcium chloride- mediated transformation, transduction, conjugation, triparental mating, DEAE, dextran- mediated transfection, infection, membrane fusion with liposomes, high velocity bombardment with DNA-coated microprojectiles, direct microinjection into single cells, electroporation, or combinations thereof can be used to introduce a nucleic acid provided herein into a host cell (see, e.g., Sambrook et al., Molecular Biology: A Laboratory Manual, Cold Spring Harbor Laboratory, NY (1989); Davis et al., Basic Methods in Molecular Biology (1986); and Neumann et al., EMBO J., 1:841 (1982)).
  • cells such as T cells, stem cells (e.g., induced pluripotent stem cells or mesenchymal stem cells), or NK cells can be designed to express one or more nucleic acids encoding a CAR described herein.
  • a population of T cells can be infected with viral vectors designed to express nucleic acid encoding a CAR described herein (e.g., a CAR having the ability to bind to a follistatin polypeptide).
  • cells such as T cells, stem cells (e.g., induced pluripotent stem cells or mesenchymal stem cells), or NK cells can be designed to express one or more nucleic acids encoding a cell engager described herein.
  • a population of T cells can be infected with viral vectors designed to express nucleic acid encoding a cell engager described herein (e.g., a cell engager having the ability to bind to a follistatin polypeptide).
  • a cell engager described herein e.g., a cell engager having the ability to bind to a follistatin polypeptide.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • a method that includes (a) introducing nucleic acid encoding the polypeptide into a host cell; (b) culturing the host cell in culture medium under conditions sufficient to express the polypeptide; (c) harvesting the polypeptide from the cell or culture medium; and (d) purifying the polypeptide (e.g., to reach at least 50, 60, 70, 80, 90, 95, 97, 98, or 99 percent purity).
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, cell engager, and/or ADC
  • a nucleic acid provided herein e.g., nucleic acid encoding an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager provided herein
  • a vector provided herein e.g., a viral vector designed to express an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager provided herein
  • a host cell e.g., a host cell designed to express an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager provided herein
  • a pharmaceutical composition for administration to a mammal (e.g., a human) having cancer to treat that mammal.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, cell engager, and/or ADC
  • a nucleic acid provided herein e.g., nucleic acid encoding an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager provided herein
  • a vector provided herein e.g., a viral vector designed to express an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager provided herein
  • a host cell e.g., a host cell designed to express an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager provided herein
  • a pharmaceutical composition for administration to a mammal (e.g., a human) to reduce the number of cancer cells within the mammal and/or to increase the survival of the mammal suffering from cancer.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, cell engager, and/or ADC
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a pharmaceutical composition for administration to a mammal e.g., a human
  • a pharmaceutically acceptable carrier such as a buffer, a salt, a surfactant, a sugar, a tonicity modifier, or combinations thereof as, for example, described elsewhere (Gervasi et al., Eur. J.
  • Examples of pharmaceutically acceptable carriers that can be used to make a pharmaceutical composition provided herein include, without limitation, water, lactic acid, citric acid, sodium chloride, sodium citrate, sodium succinate, sodium phosphate, a surfactant (e.g., polysorbate 20, polysorbate 80, or poloxamer 188), dextran 40, or a sugar (e.g., sorbitol, mannitol, sucrose, dextrose, or trehalose), or combinations thereof.
  • a surfactant e.g., polysorbate 20, polysorbate 80, or poloxamer 188
  • dextran 40 e.g., sorbitol, mannitol, sucrose, dextrose, or trehalose
  • a pharmaceutical composition designed to include a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, cell engager, and/or ADC
  • a nucleic acid, a vector, or a host cell provided herein can be formulated to include a buffer (e.g., an acetate, citrate, histidine, succinate, phosphate, or hydroxymethylaminomethane (Tris) buffer), a surfactant (e.g., polysorbate 20, polysorbate 80, or poloxamer 188), and a sugar such as sucrose.
  • a buffer e.g., an acetate, citrate, histidine, succinate, phosphate, or hydroxymethylaminomethane (Tris) buffer
  • Tris hydroxymethylaminomethane
  • a surfactant e.g., polysorbate
  • ingredients that can be included within a pharmaceutical composition provided herein include, without limitation, amino acids such as glycine or arginine, antioxidants such as ascorbic acid, methionine, or ethylenediaminetetraacetic acid (EDTA), anticancer agents such as enzalutamide, imanitib, gefitinib, erlotini, sunitinib, lapatinib, nilotinib, sorafenib, temsirolimus, everolimus, pazopanib, crizotinib, ruxolitinib, axitinib, bosutinib, cabozantinib, ponatinib, regorafenib, ibrutinib, trametinib, perifosine, bortezomib, carfilzomib, batimastat, ganetespib, obatoclax, navi
  • a pharmaceutical composition provided herein can be formulated to include one or more binders (e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cells designed to express a CAR having the ability to bind to a follistatin polypeptide, one or more cell engagers, and/or one or more ADCs) provided herein in combination with one or more checkpoint inhibitors such as anti-PD-1 antibodies or PD-1 inhibitors (e.g., cemiplimab, nivolumab, pembrolizumab, JTX-4014, spartalizumab, camrelizumab, sintilimab, tislelizumab, toripalimab, dostarlimab, INCMGA00012, AMP-224, or AMP-514), anti-PD-L1 antibodies or PD-L1 inhibitors Attorney Docket No.48881-0047WO1 / 05958 (e.g.,
  • a pharmaceutical composition provided herein can be formulated to include one or more binders (e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cells designed to express a CAR having the ability to bind to a follistatin polypeptide, one or more cell engagers, and/or one or more ADCs) provided herein in combination with one or more anticancer agents.
  • binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cells designed to express a CAR having the ability to bind to a follistatin polypeptide, one or more cell engagers, and/or one or more ADCs
  • one or more binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cells designed to express a CAR having the ability to bind to a follistatin polypeptide, one or more cell engagers, and/or one or more ADCs
  • binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cells designed to express a CAR having the ability to bind to a follistatin polypeptide, one or more cell engagers, and/or one or more ADCs
  • anticancer agents examples include, without limitation, taxol compounds, cisplatin, Monomethyl Auristatin E (MMAE), tubulin inhibitor DM4, active metabolite SN-38, and pyrrolobenzodiazepine (PDB) dimer.
  • a pharmaceutical composition is formulated to include one or more binders (e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cells designed to express a CAR having the ability to bind to a follistatin polypeptide, one or more cell engagers, and/or one or more ADCs) provided herein, any appropriate concentration of the binder can be used.
  • a pharmaceutical composition provided herein can be formulated to be a liquid that includes from about 1 mg to about 500 mg (e.g., from about 1 mg to about 500 mg, from about 10 mg to about 500 mg, from about 50 mg to about 500 mg, from about 100 mg to about 500 mg, from about 0.5 mg to about 250 mg, from about 0.5 mg to about 150 mg, from about 0.5 mg to about 100 mg, from about 0.5 mg to about 50 mg, from about 1 mg to about 300 mg, from about 2 mg to about 200 mg, from about 10 mg to about 300 mg, from about 25 mg to about 300 mg, from about 50 mg to about 150 mg, or from about 150 mg to about 300 mg) of a binder (e.g., an antibody, antigen binding fragment, antibody domain, CAR + cell population, cell engager, and/or ADC) provided herein per mL.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR + cell population, cell engager, and/or ADC
  • a pharmaceutical composition provided herein can be Attorney Docket No.48881-0047WO1 / 05958 formulated to be a solid or semi-solid that includes from about 0.5 mg to about 500 mg (e.g., from about 1 mg to about 500 mg, from about 10 mg to about 500 mg, from about 50 mg to about 500 mg, from about 100 mg to about 500 mg, from about 0.5 mg to about 250 mg, from about 0.5 mg to about 150 mg, from about 0.5 mg to about 100 mg, from about 0.5 mg to about 50 mg, from about 1 mg to about 300 mg, from about 10 mg to about 300 mg, from about 25 mg to about 300 mg, from about 50 mg to about 150 mg, or from about 150 mg to about 300 mg) of a binder (e.g., an antibody, antigen binding fragment, antibody domain, cell engager, and/or ADC) provided herein.
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, cell engager, and/or ADC
  • a pharmaceutical composition containing a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a titer of the binder being from about 1 x 10 5 to about 1 x 10 12 (e.g., from about 1 x 10 5 to about 1 x 10 10 , from about 1 x 10 5 to about 1 x 10 8 , from about 1 x 10 6 to about 1 x 10 12 , from about 1 x 10 6 to about 1 x 10 12 , from about 1 x 10 8 to about 1 x 10 12 , from about 1 x 10 9 to about 1 x 10 12 , from about 1 x 10 6 to about 1 x 10 11 , or from about 1 x 10 7 to about 1 x 10 10 ).
  • nucleic acids e.g., vectors such as viral vectors
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, CAR, and/or cell engager
  • any appropriate concentration of the nucleic acid can be used.
  • a pharmaceutical composition provided herein can be formulated to be a liquid that includes from about 0.5 mg to about 500 mg (e.g., from about 1 mg to about 500 mg, from about 10 mg to about 500 mg, from about 50 mg to about 500 mg, from about 100 mg to about 500 mg, from about 0.5 mg to about 250 mg, from about 0.5 mg to about 150 mg, from about 0.5 mg to about 100 mg, from about 0.5 mg to about 50 mg, from about 1 mg to about 300 mg, from about 2 mg to about 200 mg, from about 10 mg to about 300 mg, from about 25 mg to about 300 mg, from about 50 mg to about 150 mg, or from about 150 mg to about 300 mg) of a nucleic acid provided herein per mL.
  • a nucleic acid provided herein per mL.
  • a pharmaceutical composition provided herein can be formulated to be a solid or semi-solid that includes from about 0.5 mg to about 500 mg (e.g., from about 1 mg to about 500 mg, from about 10 mg to about 500 mg, from about 50 mg to about 500 Attorney Docket No.48881-0047WO1 / 05958 mg, from about 100 mg to about 500 mg, from about 0.5 mg to about 250 mg, from about 0.5 mg to about 150 mg, from about 0.5 mg to about 100 mg, from about 0.5 mg to about 50 mg, from about 1 mg to about 300 mg, from about 10 mg to about 300 mg, from about 25 mg to about 300 mg, from about 50 mg to about 150 mg, or from about 150 mg to about 300 mg) of a nucleic acid provided herein.
  • a nucleic acid provided herein.
  • a pharmaceutical composition designed to include a binder e.g., an antibody, antigen binding fragment, antibody domain, cell engager, and/or ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, cell engager, and/or ADC
  • agents capable of reducing aggregation of the binder when formulated include, without limitation, methionine, arginine, lysine, aspartic acid, glycine, glutamic acid, and combinations thereof.
  • one or more of these amino acids can be included within the formulation at a concentration from about 0.5 mM to about 145 mM (e.g., from about 1 mM to about 145 mM, from about 10 mM to about 145 mM, from about 100 mM to about 145 mM, from about 0.5 mM to about 125 mM, from about 0.5 mM to about 100 mM, from about 0.5 mM to about 75 mM, or from about 10 mM to about 100 mM).
  • a pharmaceutical composition provided herein can be in any appropriate form.
  • a pharmaceutical composition provided herein can designed to be a liquid, a semi-solid, or a solid.
  • a pharmaceutical composition provided herein can be a liquid solution (e.g., an injectable and/or infusible solution), a dispersion, a suspension, a tablet, a pill, a powder, a microemulsion, a liposome, or a suppository.
  • a pharmaceutical composition provided herein can be lyophilized.
  • a pharmaceutical composition provided herein e.g., a pharmaceutical composition that includes one or more binders (e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs) provided herein can be formulated with a carrier or coating designed to protect against rapid release.
  • a pharmaceutical composition provided herein can be formulated as a controlled release formulation or as a regulated release formulation as described elsewhere (U.S. Patent Application Publication Nos. 2019/0241667; 2019/0233522; and 2019/0233498).
  • a composition e.g., a pharmaceutical composition provided herein
  • binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a nucleic acid, vector, or host cell e.g., CAR + cells
  • a composition e.g., a pharmaceutical composition provided herein
  • one or more binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a nucleic acid, vector, and/or host cell e.g., CAR + cells
  • a mammal e.g., a human having cancer to treat that mammal.
  • a composition e.g., a pharmaceutical composition provided herein
  • one or more binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a nucleic acid, vector, and/or host cell e.g., CAR + cells
  • any appropriate cancer can be treated using a composition (e.g., a pharmaceutical composition provided herein) containing one or more binders (e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs) provided herein (or a nucleic acid, vector, or host cell (e.g., CAR + cells) provided herein).
  • a composition e.g., a pharmaceutical composition provided herein
  • binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a nucleic acid, vector, or host cell e.g., CAR + cells
  • a mammal e.g., a human having cancer can be treated by administering a composition (e.g., a pharmaceutical composition) containing one or more binders (e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs) provided herein to that mammal.
  • a composition e.g., a pharmaceutical composition
  • binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • cancers that can be treated as described herein include, without limitation, ovarian cancer, lung cancer, prostate cancer, pancreatic cancer, renal cancer, head and neck cancer, liver cancer, cervical cancer, urothelial cancer, thymic epithelial tumors, and breast cancer.
  • a mammal e.g., a human having a follistatin + cancer (e.g., a follistatin + Attorney Docket No.48881-0047WO1 / 05958 ovarian cancer, a follistatin + lung cancer, or a follistatin + prostate cancer)
  • a composition e.g., a pharmaceutical composition
  • one or more binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a composition provided herein e.g., a pharmaceutical composition containing one or more binders provided herein such as one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs provided herein
  • a mammal e.g., a human
  • intravenously e.g., via an intravenous injection or infusion
  • intratumorally e.g., via an intratumoral injection
  • subcutaneously e.g., via a subcutaneous injection
  • intraperitoneally e.g., via an intraperitoneal injection
  • intramuscularly e.g., via intramuscular injection.
  • the route and/or mode of administration of a composition can be adjusted for the mammal being treated.
  • an effective amount of a composition containing one or more binders (e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs) provided herein (or a nucleic acid, vector, or host cell (e.g., CAR + cells) provided herein) (e.g., a pharmaceutical composition provided herein) can be an amount that reduces the number of cancer cells within a mammal having cancer without producing significant toxicity to the mammal.
  • an effective amount of a composition containing one or more binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a nucleic acid, vector, or host cell e.g., CAR + cells
  • a pharmaceutical composition provided herein can be an amount that increases the survival time of a mammal having cancer as compared to a control mammal having Attorney Docket No.48881-0047WO1 / 05958 comparable cancer and not treated with the composition.
  • an effective amount of a binder e.g., an antibody, antigen binding fragment, antibody domain, cell engager, and/or ADC
  • a binder e.g., an antibody, antigen binding fragment, antibody domain, cell engager, and/or ADC
  • an effective amount of a binder can be from about 0.001 mg/kg to about 100 mg/kg (e.g., from about 0.001 mg/kg to about 90 mg/kg, from about 0.001 mg/kg to about 80 mg/kg, from about 0.001 mg/kg to about 70 mg/kg, from about 0.001 mg/kg to about 60 mg/kg, from about 0.001 mg/kg to about 50 mg/kg, from about 0.001 mg/kg to about 40 mg/kg, from about 0.001 mg/kg to about 30 mg/kg, from about 0.005 mg/kg to about 100 mg/kg, from about 0.01 mg/kg to about 100 mg/kg, from about 0.05 mg/kg to about 100 mg/kg, from about 0.1 mg/kg to
  • the effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment.
  • Various factors can influence the actual effective amount used for a particular application. For example, the severity of cancer when treating a mammal having cancer, the route of administration, the age and general health condition of the mammal, excipient usage, the possibility of co-usage with other therapeutic or prophylactic treatments such as use of other agents (e.g., checkpoint inhibitors), and the judgment of the treating physician may require an increase or decrease in the actual effective amount of a composition provided herein (e.g., a pharmaceutical composition containing one or more binders provided herein) that is administered.
  • a composition provided herein e.g., a pharmaceutical composition containing one or more binders provided herein
  • an effective frequency of administration of a composition containing one or more binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a nucleic acid, vector, or host cell e.g., CAR + cells
  • a pharmaceutical composition provided herein can be a frequency that reduces the number of cancer cells within a mammal having cancer without producing significant toxicity to the mammal.
  • an effective frequency of administration of a composition containing one or more binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a nucleic acid, vector, or host cell e.g., CAR + cells
  • a pharmaceutical composition provided herein can be a frequency that increases the survival time of a mammal having cancer as compared to a control mammal having comparable cancer and not treated with the composition.
  • an effective frequency of administration of a pharmaceutical composition provided herein such as a pharmaceutical composition containing one or more binders provided herein can be from about twice daily to about once a year (e.g., from about twice daily to about once a month, from about twice daily to about once a week, from about once daily to about once a month, or from one once daily to about once a week).
  • the frequency of administration of a pharmaceutical composition provided herein such as a pharmaceutical composition containing one or more binders provided herein can be daily.
  • the frequency of administration of a pharmaceutical composition provided herein such as a pharmaceutical composition containing one or more binders provided herein can remain constant or can be variable during the duration of treatment. Various factors can influence the actual effective frequency used for a particular application.
  • the severity of the cancer, the route of administration, the age and general health condition of the mammal, excipient usage, the possibility of co-usage with other therapeutic or prophylactic treatments such as use of other agents (e.g., checkpoint inhibitors), and the judgment of the treating physician may require an increase or decrease in the actual effective Attorney Docket No.48881-0047WO1 / 05958 frequency of administration of a composition provided herein (e.g., a pharmaceutical composition containing one or more binders provided herein).
  • a composition provided herein e.g., a pharmaceutical composition containing one or more binders provided herein.
  • an effective duration of administration of a composition containing one or more binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a nucleic acid, vector, or host cell e.g., CAR + cells
  • a pharmaceutical composition provided herein can be a duration that reduces the number of cancer cells within a mammal without producing significant toxicity to the mammal.
  • an effective duration of administration of a composition containing one or more binders e.g., one or more antibodies, one or more antigen binding fragments, one or more antibody domains, one or more cell engagers, and/or one or more ADCs
  • a nucleic acid, vector, or host cell e.g., CAR + cells
  • a pharmaceutical composition provided herein can be a duration that increases the survival time of a mammal having cancer as compared to a control mammal having comparable cancer and not treated with the composition.
  • an effective duration of administration of a pharmaceutical composition provided herein can vary from a single time point of administration to several weeks to several months (e.g., 4 to 12 weeks). Multiple factors can influence the actual effective duration used for a particular application. For example, the severity of the cancer, the route of administration, the age and general health condition of the mammal, excipient usage, the possibility of co-usage with other therapeutic or prophylactic treatments such as use of other agents (e.g., checkpoint inhibitors), and the judgment of the treating physician may require an increase or decrease in the actual effective duration of administration of a composition provided herein (e.g., a pharmaceutical composition containing one or more binders provided herein).
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a label e.g., a covalently attached radioactive, enzymatic, colorimetric, or fluorescent label.
  • the labelled binder can be used to detect the presence or absence of a follistatin polypeptide (e.g., a human follistatin polypeptide) within a biological sample in vitro.
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • biological samples that can be assessed using a binder (e.g., an antibody, antigen binding fragment, and/or antibody domain) provided herein include, without limitation, serum samples, plasma samples, tissue samples, biopsy samples, cell line samples, and tissue culture samples.
  • a biological sample that can be assessed as described herein can include mammalian body tissues and/or cells such as leukocytes, ovary tissue or cells, prostate tissue or cells, heart tissue or cells, placenta tissue or cells, pancreas tissue or cells, liver tissue or cells, spleen tissue or cells, lung tissue or cells, breast tissue or cells, head and neck tissue or cells, endometrium tissue or cells, colon tissue or cells, colorectal tissue or cells, cervix tissue or cells, stomach tissue or cells, or umbilical tissue or cells that may express a follistatin polypeptide (e.g., a human follistatin polypeptide).
  • mammalian body tissues and/or cells such as leukocytes, ovary tissue or cells, prostate tissue or cells, heart tissue or cells, placenta tissue or cells, pancreas tissue or cells, liver tissue or cells, spleen tissue or cells, lung tissue or cells, breast tissue or cells, head and neck tissue or cells, endometrium tissue or
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • cell sorting e.g., fluorescence activated cell sorting
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a label e.g., a covalently attached radioactive label
  • a follistatin polypeptide e.g., a human follistatin polypeptide
  • a mammal e.g., a human
  • a binder e.g., an antibody, antigen binding fragment, and/or antibody domain
  • a mammal e.g., a human
  • a mammal e.g., a human
  • a mammal Attorney Docket No.48881-0047WO1 / 05958 can be scanned to evaluate the location(s) of a labelled binder provided herein within the mammal.
  • the mammal can be imaged using NMR or other tomographic techniques.
  • radiolabels such as 131 I, 111 In, 123 I, 99m Tc, 32 P, 33 P, 125 I, 3 H, 14 C, and 188 Rh
  • fluorescent labels such as fluorescein and rhodamine
  • a polypeptide having amino acid residues 1 to 334 of human follistatin polypeptide set forth in Example 2 was fused to the AviTag/His tag/Fc tag sequence at the C-terminus of the follistatin sequence, and the follistatin-AviTag polypeptide was used for panning of human VH domain phage-displayed libraries.
  • 14 VH domains (Clones #1-#14; Examples 3-16) were identified.
  • the 14 VH domains (Clones #1-#14) were assessed for binding affinity and specificity to a human follistatin polypeptide using an ELISA ( Figure 4).
  • Clones #1-#14 exhibited high affinity binding to a human follistatin polypeptide having EC 50 (nM) values of 17, 19, 2.5, 18, 3, 2.5, 3.5, 3.2, 12.3, 0.5, 21, 21, 68, and 23, respectively.
  • 12 of Attorney Docket No.48881-0047WO1 / 05958 the 14 VH domains were used to create Fc versions of the binders and assessed for binding to a human follistatin polypeptide using an ELISA ( Figure 5). 12 of the 14 VH domains were assessed for binding to BSA ( Figure 6).
  • FST3G9 and FST1E8 were used to test if anti-follistatin binders can enhance the cancer cell killing by chemotherapeutic agents such as cisplatin.
  • Exemplary human IgG1-derived hinge EPKSCDKTHTCPPCP (SEQ ID NO:172) Attorney Docket No.48881-0047WO1 / 05958
  • Exemplary human IgG2-derived hinge DKTHTCPPCPAPPVA (SEQ ID NO:173)
  • Exemplary human IgG4-derived hinges ESKYGPPCPPCP (SEQ ID NO:174) ESKYGPPCPSCP (SEQ ID NO:175)
  • Exemplary human CD8 ⁇ -derived hinge KPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIY (SEQ ID NO:176) TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIY (SEQ ID NO:177) TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIY (SEQ ID NO:177) TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (
  • the nucleic acid encoding these CARs under the control of a CMV promoter are introduced into human T cells.
  • CAR-expressing T cells for the CAR clones are used as effector cells to kill target cells containing follistatin on their cell surface such as follistatin + cancer cells within a human.
  • Example 34 – Designing BiKEs from binders having the ability to bind to a human follistatin polypeptide Clones #1-#14 are used to make BiKEs having the following configuration: VH Domain of any one of Clones #1-#14 + (G4S) linker (SEQ ID NO:158) + Anti-NKG2A VH + linker + Anti-NKG2A VL.
  • BiKEs are used to direct NK cells to kill cells containing human follistatin on their cell surface such as follistatin + cancer cells.
  • Example 35 – Designing BiTEs from binders having the ability to bind to a human follistatin polypeptide Clones #1-#14 are used to make BiTEs having the following configuration: VH Domain of any one of Clones #1-#14 + (GSGSS) 3 linker (SEQ ID NO:650) + Anti-CD3 scFv + GSGSSG linker (SEQ ID NO:649) + hIgG1 Fc.
  • Such BiTEs are used to direct T cells to kill cells containing human follistatin on their cell surface such as follistatin + cancer cells.

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  • Proteomics, Peptides & Aminoacids (AREA)
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Abstract

Ce document concerne des procédés et matériels impliqués dans la liaison d'un liant (par exemple, un anticorps, un fragment de liaison à l'antigène, un domaine d'anticorps, un récepteur antigénique chimérique, un anticorps engageant les cellules et/ou un conjugué anticorps-médicament) à un polypeptide de follistatine. Par exemple, l'invention concerne des liants (par exemple, des anticorps, des fragments de liaison à l'antigène, des domaines d'anticorps, des récepteurs antigéniques chimériques, des anticorps engageant les cellules et/ou des ADC) qui se lient à un polypeptide de follistatine ainsi que des procédés et matériels permettant d'utiliser une ou plusieurs molécules de liaison de ce type pour traiter un mammifère (par exemple, un être humain) atteint d'un cancer.
PCT/US2024/013973 2023-02-02 2024-02-01 Molécules se liant à des polypeptides de follistatine Ceased WO2024163723A2 (fr)

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AU2024213820A AU2024213820A1 (en) 2023-02-02 2024-02-01 Molecules that bind to follistatin polypeptides

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