WO2022132929A2 - Molécules d'anticorps multispécifiques et leurs utilisations - Google Patents

Molécules d'anticorps multispécifiques et leurs utilisations Download PDF

Info

Publication number
WO2022132929A2
WO2022132929A2 PCT/US2021/063558 US2021063558W WO2022132929A2 WO 2022132929 A2 WO2022132929 A2 WO 2022132929A2 US 2021063558 W US2021063558 W US 2021063558W WO 2022132929 A2 WO2022132929 A2 WO 2022132929A2
Authority
WO
WIPO (PCT)
Prior art keywords
seq
amino acid
acid sequence
antibody
antibody molecule
Prior art date
Application number
PCT/US2021/063558
Other languages
English (en)
Other versions
WO2022132929A3 (fr
Inventor
Allen Ebens
Original Assignee
Vera Therapeutics, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Vera Therapeutics, Inc. filed Critical Vera Therapeutics, Inc.
Publication of WO2022132929A2 publication Critical patent/WO2022132929A2/fr
Publication of WO2022132929A3 publication Critical patent/WO2022132929A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2809—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
    • 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
    • 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
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/75—Agonist effect on antigen
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/77—Internalization into the cell

Definitions

  • Hematopoietic stem cell transplantation is a standard therapy for various non- malignant hematological.
  • a conditioning therapy comprising myeloablative doses of chemotherapy and/or radiation to make the recipient suitable for engraftment of the donor stem cells.
  • This conditioning therapy is often accompanied by severe complications, including neutropenia and thrombocytopenia. Managing these adverse events is critical to minimizing any life-threatening complications. Accordingly, new conditioning therapies are needed to prepare subjects for HSCT that minimize serious, adverse effects.
  • Tumor-based immunosuppression is common in many types of malignancies, including hematological cancers and solid tumors. This immunosuppression interferes with the subject’s ability mount an immune response against the cancer and reduces the efficacy of anti-cancer therapies.
  • immunomodulatory anti-cancer agents there remains an unmet need for new therapies to mitigate tumor-induced immunosuppression to enhance efficacy of anti-cancer agents and therapies and increase survival following diagnosis.
  • the disclosure relates, inter alia, to novel multispecific antibody molecules, such as bispecific antibody molecules, that include a first moiety that binds to an immune effector cell antigen (sometimes referred to herein as “immune effector cell antigen-targeting moiety”) and a second moiety that binds to a hematopoietic stem cell (HSC) antigen (sometime referred to herein as “hematopoietic stem cell (HSC) antigen-targeting moiety”).
  • an immune effector cell antigen sometimes referred to herein as “immune effector cell antigen-targeting moiety”
  • HSC hematopoietic stem cell
  • the first moiety that binds to an immune effector cell antigen can be a moiety that binds to CD3 (sometimes referred to herein as “CD3 -targeting moiety”) and the second moiety that binds to an HSC antigen can be a moiety that binds to CD117 (sometime referred to herein as “CD117- targeting moiety”).
  • the moiety that binds to CD3 can be replaced by a moiety that binds to another immune effector cell antigen.
  • the immune effector cell antigen is the T cell receptor (TCR), CD28, CD16, NKG2D, 0X40, 4-1BB, CD2,
  • the moiety that binds to CD117 can be replaced by a moiety that binds to another HSC antigen.
  • the HSC antigen is CD34.
  • the disclosure provides a multispecific antibody molecule, comprising a first moiety that binds to CD3; and a second moiety that binds to CD117.
  • the first moiety binds to CD3s.
  • the multispecific molecule further comprises a third moiety.
  • the third moiety binds to an HSC antigen.
  • the HSC antigen comprises CD34.
  • the HSC antigen comprises CD117.
  • the third moiety binds to an immune effector cell antigen.
  • the immune effector cell antigen comprises CD3, the TCR, CD28, CD16, NKG2D, 0X40, 4- 1BB, CD2, CD5, or CD95.
  • the first moiety that binds to CD3 comprises (a) one, two, or all of HCDR1, HCDR2, or HCDR3 of an anti-CD3 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions); (b) one, two, or all of LCDR1, LCDR2, or LCDR3 of an anti-CD3 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions); or (c) both (a) and (b).
  • the first moiety that binds to CD3 further comprises: (a) a VH of an anti-CD3 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions); (b) a VL of an anti-CD3 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions); or (c) both (a) and (b).
  • the first moiety that binds to CD3 further comprises: (a) a heavy chain of an anti-CD3 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions); (b) a light chain of an anti-CD3 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions) or (c) both (a) and (b).
  • the second moiety that binds to CD117 comprises:
  • first moiety that binds to CD117 comprises: (a) a VH of an anti- CD117 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions); (b) a VL of an anti- CD117 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions) or (c) both (a) and (b).
  • the first moiety that binds to CD117 comprises: (a) a heavy chain of an anti- CD117 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions); (b) a light chain of an anti- CD117 antibody described herein (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions) or (c) both (a) and (b).
  • binding of the first moiety to a cell expressing CD3 modulates a function of the cell expressing CD3 (e.g., formation of an immunologic synapse with target cells, cellular activation, proliferation, and/or target cell killing that, in some instances, is mediated by perforin and granzyme release).
  • the cell expressing CD3 is an immune cell, e.g., an effector cell.
  • binding of the second moiety to a cell expressing CD117 modulates a function of the cell expressing CD117 (e.g., inhibits or decreases or fails to activate mast cell degranulation).
  • the cell expressing CD117 is a hematopoietic stem cell (HSC), a multipotent progenitor (MPP), a common myeloid progenitor (CMP), or a cancer cell (e.g., a gastrointestinal stromal tumor (GIST) cancer cell or an acute myeloid leukemia (AML) cancer cell).
  • HSC hematopoietic stem cell
  • MPP multipotent progenitor
  • CMP common myeloid progenitor
  • cancer cell e.g., a gastrointestinal stromal tumor (GIST) cancer cell or an acute myeloid leukemia (AML) cancer cell.
  • the multispecific antibody molecule further comprises an immunoglobulin chain constant region (e.g., Fc region) chosen from, e.g., the heavy chain constant regions of IgGl, IgG2, IgG3, or IgG4, more particularly, the heavy chain constant region of human IgGl, IgG2, IgG3, or IgG4.
  • an immunoglobulin chain constant region e.g., Fc region
  • the immunoglobulin chain constant region (e.g., Fc region) is altered, e.g., mutated, to increase, decrease, or otherwise modulate one or more of: Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector function, or complement function.
  • the immunoglobulin chain constant region is mutated to decrease an effector function.
  • the mutation is a substitution mutation at position N297 (EU numbering).
  • the mutation is a N297G substitution mutation.
  • the immunoglobulin chain constant region is mutated to decrease FcRn binding.
  • the mutation comprises one or more substitution mutations at one or more of amino acid residues selected from the group consisting of Ile253, Ser254, His435, Tyr436, Glu233-Gly236, Arg255, Lys288, Ser415, and His433.
  • the mutation comprises one or more substitution mutations at one or more of amino acid residues selected from the group consisting of Ile253, Ser254, His435, Tyr436, Glu233-Gly236, Arg255, Lys288, Ser415, His310, and His433.
  • the one or more substitution mutations are one or more alanine substitution mutations.
  • the multispecific antibody molecule comprises a dimerization module between the first moiety and the second moiety.
  • the dimerization module comprises an immunoglobulin constant domain, e.g., a heavy chain constant domain (e.g., a homodimeric or heterodimeric heavy chain constant region, e.g., an Fc region), or a constant domain of an immunoglobulin variable region (e.g., a Fab region).
  • the dimerization module comprises one or more immunoglobulin chain constant regions (e.g., Fc regions) comprising one or more of: a paired cavity-protuberance (“knob-in-a hole”), an electrostatic interaction, or a strand-exchange.
  • the one or more immunoglobulin chain constant regions comprise an amino acid substitution at a position chosen from one or more of 347, 349, 350, 351, 366, 368, 370, 392, 394, 395, 397, 398, 399, 405, 407, or 409, e.g., of the Fc region of human IgGl, optionally wherein the one or more immunoglobulin chain constant regions (e.g., Fc regions) comprise an amino acid substitution chosen from: T366S, E368A, or Y407V (e.g., corresponding to a cavity or hole), or T366W (e.g., corresponding to a protuberance or knob), or a combination thereof.
  • T366S, E368A, or Y407V e.g., corresponding to a cavity or hole
  • T366W e.g., corresponding to a protuberance or knob
  • the multispecific antibody molecule further comprises a linker, e.g., a linker between the first moiety that binds to CD3 and the second moiety that binds to CD117.
  • the linker is chosen from: a cleavable linker, a non-cleavable linker, a peptide linker, a flexible linker, a rigid linker, a helical linker, or a non-helical linker.
  • the linker is a peptide linker.
  • the peptide linker comprises Gly and Ser, or an amino acid linker sequence described herein.
  • the disclosure provides one or more nucleic acids encoding the multispecific antibody molecule as described herein or a functional portion thereof.
  • the disclosure provides one or more vectors, e.g., one or more expression vectors, comprising one or more nucleic acids encoding a multispecific antibody as described herein.
  • the disclosure provides one or more cells, e.g., one or more host cells, comprising the one or more nucleic acids encoding a multispecific antibody as described herein or the one or more vectors encoding a multispecific antibody as described herein.
  • the disclosure provides a method of making, e.g., producing, the multispecific antibody molecule as described herein, comprising culturing one or more host cells described herein, under suitable conditions, e.g., conditions suitable for gene expression and/or homo- or heterodimerization.
  • the disclosure provides a composition, e.g., a pharmaceutical composition, comprising the multispecific antibody molecule as described herein, optionally further comprising a pharmaceutically acceptable carrier, excipient, or stabilizer.
  • the disclosure provides a kit comprising the multispecific antibody molecule as described herein, and instructions to use the multispecific antibody molecule.
  • the disclosure provides a method of preparing a subject for a hematopoietic stem cell transplantation (HSCT), comprising administering to a subject in need thereof an effective amount of the multispecific antibody molecule or the composition as described herein, thereby preparing the subject.
  • HSCT hematopoietic stem cell transplantation
  • the disclosure provides a multispecific antibody molecule as described herein or a pharmaceutical composition thereof for use in a method of preparing a subject for a hematopoietic stem cell transplantation (HSCT), the method comprising administering to a subject in need thereof an effective amount of the multispecific antibody molecule or pharmaceutical composition thereof, thereby preparing the subject.
  • HSCT hematopoietic stem cell transplantation
  • the subject has a blood disorder.
  • the blood disorder is a non-malignant blood disorder.
  • the blood disorder is thalassemia or sickle cell disease.
  • the subject has, or is determined to have, an autoimmune disorder.
  • the autoimmune disorder is lupus, rheumatoid, arthritis (RA), multiple sclerosis (MS), type I diabetes, or Crohn’s disease.
  • the HSCT is an autologous HSCT. In other embodiments, the HSCT is an allogeneic HSCT.
  • the disclosure provides a method of treating a disorder in a subject, comprising administering to the subject an effective amount of the multispecific antibody molecule or the composition as described herein, wherein the subject is in need of receiving an HSCT, thereby treating the disorder.
  • the disclosure provides a multispecific antibody molecule as described herein or a pharmaceutical composition thereof for use in a method of treating a disorder in a subject, the method comprising administering to the subject an effective amount of the multispecific antibody molecule or pharmaceutical composition thereof, wherein the subject is in need of receiving an HSCT, thereby treating the disorder.
  • the disclosure provides a method of treating a disorder, comprising administering to the subject in need thereof an effective amount of the multispecific antibody molecule or the pharmaceutical composition as described herein, and an HSCT, thereby treating the disorder.
  • the disclosure provides a multispecific antibody molecule as described herein or a pharmaceutical composition thereof for use in a method of treating a disorder, the method comprising administering to the subject in need thereof an effective amount of the multispecific antibody molecule, and an HSCT, thereby treating the disorder.
  • the disorder is a blood disorder. In some embodiments, the disorder is an autoimmune disorder.
  • the subject exhibits one or more of (a) reduced anemia and thrombocytopenia; (b) reduced organ damage; (c) reduced sterility; (d) reduced endocrine dysfunction; (e) reduced cognitive decline; (f) reduced secondary malignancies; (g) reduced mortality; (h) reduced infections; (i) reduced neutropenia; and/or (j) reduced reactivation of latent viruses (e.g., EBV and CMV) as compared to a subject receiving myeloablative chemotherapy or radiation therapy.
  • latent viruses e.g., EBV and CMV
  • the disclosure provides a method of reducing immunosuppression, comprising administering to a subject in need thereof an effective amount of the multispecific antibody molecule or the pharmaceutical composition as described herein, thereby reducing immunosuppression.
  • the disclosure provides a multispecific antibody molecule as described herein or a pharmaceutical composition thereof for use in a method of reducing immunosuppression, the method comprising administering to a subject in need thereof an effective amount of the multispecific antibody molecule or the pharmaceutical composition thereof, thereby reducing immunosuppression.
  • the subject is receiving, or has received, a cancer therapy. In some embodiments, the subject has, or is at risk of having, a cancer.
  • the disclosure provides a method of treating a cancer, comprising administering to a subject in need thereof an effective amount of the multispecific antibody molecule or the pharmaceutical composition as described herein, thereby treating the cancer.
  • the disclosure provides a multispecific antibody molecule as described herein or a pharmaceutical composition thereof for use in a method of treating a cancer, the method comprising administering to a subject in need thereof an effective amount of the multispecific antibody molecule or the pharmaceutical composition, thereby treating the cancer
  • the subject has, or is identified as having, immunosuppression.
  • the multispecific antibody molecule is administered responsive to a determination of immunosuppression in the subject.
  • the cancer is a solid tumor cancer.
  • the cancer is a hematological cancer, optionally wherein the hematological cancer is a leukemia, a lymphoma, or a myeloma.
  • the cancer is GIST. In some embodiments, the cancer is AML. In some embodiments, the cancer is a seminoma. In some embodiments, the cancer is an adenoid- cystic carcinoma. In some embodiments, the cancer is a malignant melanoma. In some embodiments, the cancer is a large-cell carcinoma of the lung. In some embodiments, the cancer has a CD117 mutation, e.g., an oncogenic CD117 mutation. In some embodiments, the CD117 mutation is a gain of function mutation.
  • the methods further comprise administering to the subject a second therapy.
  • the second therapy comprises a chemotherapy, a radiotherapy
  • the second therapy is administered prior to, concurrent with, or after administration of the multispecific antibody molecule.
  • multispecific antibody molecules that include, for example, a plurality of (e.g., two or more) functionalities (or binding specificities), comprising a first moiety that binds to an immune cell antigen (e.g., an immune effector cell antigen) and a second moiety that binds to a hematopoietic stem cell (HSC) antigen or a cancer cell antigen.
  • an immune cell antigen e.g., an immune effector cell antigen
  • HSC hematopoietic stem cell
  • the multispecific antibody molecule comprises (i) a first moiety that binds to CD3 and (ii) a second moiety that binds to CD117.
  • the first moiety binds to a cell expressing CD3, e.g., a CD3 -expressing cell described herein.
  • the first moiety modulates one or more functions of the CD3-expressing cell.
  • the second moiety binds to a cell expressing CD117, e.g., a CD117-expressing cell described herein.
  • the second moiety modulates one or more functions of the CD117- expressing cell.
  • the multispecific antibody molecule modulates one or more functions of a cell expressing CD3 (e.g., a CD3 -expressing cell described herein) and one or more functions of a cell expressing CD117 (e.g., a CD117-expressing cell described herein).
  • the multispecific antibody molecule is a bispecific antibody molecule, a trispecific antibody molecule, or a tetraspecific antibody molecule. In an embodiment, the multispecific antibody molecule is a bispecific antibody molecule.
  • the multispecific antibody molecules disclosed herein are expected to localize (e.g., bridge) and/or activate a CD3-expressing cell (e.g., an immune cell, e.g., an immune effector cell, e.g., a T cell), in the presence of a cell expressing CD117 (e.g., a hematopoietic stem cell (HSC), a multipotent progenitor (MPP), a common myeloid progenitor (CMP), or a cancer cell (e.g., a gastrointestinal stromal tumor (GIST) cancer cell or an acute myeloid leukemia (AML) cancer cell).
  • a CD3-expressing cell e.g., an immune cell, e.g., an immune effector cell, e.g., a T cell
  • a cell expressing CD117 e.g., a hematopoietic stem cell (HSC), a multipotent progenitor (MPP), a common mye
  • compositions e.g., pharmaceutical compositions
  • kits, devices, and containers comprising the multispecific antibody molecules described herein, nucleic acids encoding the multispecific antibody molecules described herein, vectors comprising the aforesaid nucleic acids, cells comprising the aforesaid nucleic acids and vectors, methods of producing the multispecific antibody molecules described herein, and methods of using the multispecific antibody molecules described herein.
  • the term “molecule” as used in, e.g., antibody molecule, cytokine molecule, receptor molecule, includes full-length, naturally-occurring molecules, as well as variants, e.g., functional variants (e.g., truncations, fragments, mutated (e.g., substantially similar sequences) or derivatized form thereof), so long as at least one function and/or activity of the unmodified (e.g., naturally-occurring) molecule remains.
  • the articles “a” and “an” refer to one or more than one, e.g., to at least one, of the grammatical object of the article.
  • the use of the words “a” or “an” when used in conjunction with the term “comprising” herein may mean “one,” but it is also consistent with the meaning of "one or more,” “at least one,” and “one or more than one.”
  • Antibody molecule refers to a protein, e.g., an immunoglobulin chain or fragment thereof, comprising at least one immunoglobulin variable domain sequence.
  • An antibody molecule encompasses antibodies (e.g., full-length antibodies) and antibody fragments.
  • an antibody molecule comprises an antigen binding or functional fragment of a full-length antibody, or a full-length immunoglobulin chain.
  • a full-length antibody is an immunoglobulin (Ig) molecule (e.g., an IgG antibody) that is naturally occurring or formed by normal immunoglobulin gene fragment recombinatorial processes).
  • an antibody molecule refers to an immunologically active, antigen-binding portion of an immunoglobulin molecule, such as an antibody fragment.
  • An antibody fragment e.g., functional fragment, is a portion of an antibody, e.g., Fab, Fab', F(ab')2, F(ab)2, variable fragment (Fv), domain antibody (dAb), or single chain variable fragment (scFv).
  • a functional antibody fragment binds to the same antigen as that recognized by the intact (e.g., full-length) antibody.
  • antibody fragment or “functional fragment” also include isolated fragments consisting of the variable regions, such as the “Fv” fragments consisting of the variable regions of the heavy and light chains or recombinant single chain polypeptide molecules in which light and heavy variable regions are connected by a peptide linker (“scFv proteins”).
  • an antibody fragment does not include portions of antibodies without antigen binding activity, such as Fc fragments or single amino acid residues.
  • Exemplary antibody molecules include full length antibodies and antibody fragments, e.g., dAb (domain antibody), single chain, Fab, Fab’, and F(ab’)2 fragments, and single chain variable fragments (scFvs).
  • an “immunoglobulin variable domain sequence” refers to an amino acid sequence which can form the structure of an immunoglobulin variable domain.
  • the sequence may include all or part of the amino acid sequence of a naturally occurring variable domain.
  • the sequence may or may not include one, two, or more N- or C-terminal amino acids, or may include other alterations that are compatible with formation of the protein structure.
  • an antibody molecule is monospecific, e.g., it comprises binding specificity for a single epitope. In some embodiments, an antibody molecule is multi specific, e.g., it comprises a plurality of immunoglobulin variable domain sequences, where a first 10
  • an antibody molecule is a bispecific antibody molecule.
  • Bispecific antibody molecule refers to an antibody molecule that has specificity for more than one (e.g., two, three, four, or more) epitope and/or antigen.
  • Antigen refers to a molecule that can provoke an immune response, e.g., involving activation of certain immune cells and/or antibody generation. Any macromolecule, including almost all proteins or peptides, can be an antigen. Antigens can also be derived from genomic recombinant or DNA. For example, any DNA comprising a nucleotide sequence or a partial nucleotide sequence that encodes a protein capable of eliciting an immune response encodes an “antigen.” In some embodiments, an antigen does not need to be encoded solely by a full-length nucleotide sequence of a gene, nor does an antigen need to be encoded by a gene at all.
  • an antigen can be synthesized or can be derived from a biological sample, e.g., a tissue sample, a tumor sample, a cell, or a fluid with other biological components.
  • a biological sample e.g., a tissue sample, a tumor sample, a cell, or a fluid with other biological components.
  • a tumor antigen or interchangeably, a “cancer antigen” includes any molecule present on, or associated with, a cancer, e.g., a cancer cell or a tumor microenvironment that can provoke an immune response.
  • an “immune cell antigen” includes any molecule present on, or associated with, an immune cell that can provoke an immune response.
  • the “antigen-binding site,” or “binding portion” of an antibody molecule refers to the part of an antibody molecule, e.g., an immunoglobulin (Ig) molecule, that participates in antigen binding.
  • the antigen binding site is formed by amino acid residues of the variable (V) regions of the heavy (H) and light (L) chains.
  • V variable regions of the heavy and light chains
  • hypervariable regions Three highly divergent stretches within the variable regions of the heavy and light chains, referred to as hypervariable regions, are disposed between more conserved flanking stretches called “framework regions,” (FRs).
  • FRs are amino acid sequences that are naturally found between, and adjacent to, hypervariable regions in immunoglobulins.
  • the three hypervariable regions of a light chain and the three hypervariable regions of a heavy chain are disposed relative to each other in three-dimensional space to form an antigen-binding surface, which is complementary to the three-dimensional surface of a bound antigen.
  • the three hypervariable regions of each of the heavy and light chains are referred to as “complementarity-determining 11
  • CDRs The framework region and CDRs have been defined and described, e.g., in Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242, and Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917.
  • Each variable chain (e.g., variable heavy chain and variable light chain) is typically made up of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the amino acid order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.
  • cancer as used herein can encompass all types of oncogenic processes and/or cancerous growths.
  • cancer includes primary tumors as well as metastatic tissues or malignantly transformed cells, tissues, or organs.
  • cancer encompasses all histopathology and stages, e.g., stages of invasiveness/severity, of a cancer.
  • cancer includes relapsed and/or resistant cancer.
  • cancer and “tumor” can be used interchangeably. For example, both terms encompass solid and liquid tumors.
  • cancer or “tumor” includes premalignant, as well as malignant cancers and tumors.
  • an “immune cell” refers to any of various cells that function in the immune system, e.g., to protect against agents of infection and foreign matter.
  • this term includes leukocytes, e.g., neutrophils, eosinophils, basophils, lymphocytes, and monocytes.
  • leukocytes include phagocytes (e.g., macrophages, neutrophils, and dendritic cells), mast cells, eosinophils, basophils, and natural killer cells.
  • phagocytes e.g., macrophages, neutrophils, and dendritic cells
  • mast cells e.g., eosinophils, basophils, and natural killer cells.
  • Innate leukocytes identify and eliminate pathogens, either by attacking larger pathogens through contact or by engulfing and then killing microorganisms and are mediators in the activation of an adaptive immune response.
  • lymphocytes The cells of the adaptive immune system are special types of leukocytes, called lymphocytes.
  • B cells and T cells are important types of lymphocytes and are derived from hematopoietic stem cells in the bone marrow. B cells are involved in the humoral immune response, whereas T cells are involved in cell-mediated immune response.
  • immune cell includes immune effector cells.
  • Immuno effector cell refers to a cell that is involved in an immune response, e.g., in the promotion of an immune effector response.
  • immune effector cells include, but are not limited to, T cells, e.g., alpha/beta T cells and gamma/delta T cells, B cells, natural killer (NK) cells, natural killer T (NK T) cells, and mast cells.
  • effector function refers to a specialized function of a cell. Effector function of a T cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines.
  • compositions and methods of the present invention encompass polypeptides and nucleic acids having the sequences specified, or sequences substantially identical or similar thereto, e.g., sequences at least 80%, 85%, 90%, 95% identical or higher to the sequence specified.
  • substantially identical is used herein to refer to a first amino acid that contains a sufficient or minimum number of amino acid residues that are i) identical to, or ii) conservative substitutions of aligned amino acid residues in a second amino acid sequence such that the first and second amino acid sequences can have a common structural domain and/or common functional activity.
  • amino acid sequences that contain a common structural domain having at least about 80%, 85%, 90%. 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence, e.g., a sequence provided herein.
  • nucleotide sequence in the context of nucleotide sequence, the term "substantially identical" is used herein to refer to a first nucleic acid sequence that contains a sufficient or minimum number of nucleotides that are identical to aligned nucleotides in a second nucleic acid sequence such that the first and second nucleotide sequences encode a polypeptide having common functional activity, or encode a common structural polypeptide domain or a common functional polypeptide activity.
  • variant refers to a polypeptide that has a substantially identical amino acid sequence to a reference amino acid sequence or is encoded by a substantially identical nucleotide sequence. In some embodiments, the variant is a functional variant.
  • the term “functional variant” refers to a polypeptide that has a substantially identical amino acid sequence to a reference amino acid sequence, or is encoded by a substantially identical nucleotide sequence, and is capable of having one or more activities of the reference amino acid sequence.
  • the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes).
  • the length of a reference sequence aligned for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, 60%, and even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence.
  • the amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared.
  • amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology”
  • the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.
  • the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm.
  • the percent identity between two amino acid sequences is determined using the Needleman and Wunsch ((1970) J. Mol. Biol. 48:444-453 ) algorithm which has been incorporated into the GAP program in the GCG software package (available at http://www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6.
  • the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (available at http://www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6.
  • a particularly preferred set of parameters are a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.
  • the percent identity between two amino acid or nucleotide sequences can be determined using the algorithm of E. Meyers and W. Miller ((1989) CABIOS, 4:11-17) which has been incorporated into the ALIGN program (version 2.0), using a PAM 120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
  • nucleic acid and protein sequences described herein can be used as a "query sequence" to perform a search against public databases to, for example, identify other family members or related sequences.
  • Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10.
  • Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25:3389-3402.
  • the default parameters of the respective programs e.g., XBLAST and NBLAST
  • XBLAST and NBLAST See http://www.ncbi.nlm.nih.gov.
  • molecules of the present invention may have additional conservative or non-essential amino acid substitutions, which do not have a substantial effect on their functions.
  • amino acid is intended to embrace all molecules, whether natural or synthetic, which include both an amino functionality and an acid functionality and capable of being included in a polymer of naturally occurring amino acids.
  • exemplary amino acids include naturally occurring amino acids; analogs, derivatives and congeners thereof; amino acid analogs having variant side chains; and all stereoisomers of any of any of the foregoing.
  • amino acid includes both the D- or L- optical isomers and peptidomimetics.
  • a “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain.
  • Families of amino acid residues having similar side chains have been defined in the art. These families 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), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).
  • polypeptide polypeptide
  • peptide protein
  • protein if single chain
  • the terms “polypeptide”, “peptide” and “protein” are used interchangeably herein to refer to polymers of amino acids of any length.
  • the polymer may be 15
  • T2110-7026WO linear or branched it may comprise modified amino acids, and it may be interrupted by nonamino acids.
  • the terms also encompass an amino acid polymer that has been modified; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation, such as conjugation with a labeling component.
  • the polypeptide can be isolated from natural sources, can be a produced by recombinant techniques from a eukaryotic or prokaryotic host, or can be a product of synthetic procedures.
  • nucleic acid refers to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof.
  • the polynucleotide may be either single-stranded or double-stranded, and if single- stranded may be the coding strand or non-coding (antisense) strand.
  • a polynucleotide may comprise modified nucleotides, such as methylated nucleotides and nucleotide analogs.
  • the sequence of nucleotides may be interrupted by non-nucleotide components.
  • a polynucleotide may be further modified after polymerization, such as by conjugation with a labeling component.
  • the nucleic acid may be a recombinant polynucleotide, or a polynucleotide of genomic, cDNA, semisynthetic, or synthetic origin which either does not occur in nature or is linked to another polynucleotide in a non-natural arrangement.
  • isolated refers to material that is removed from its original or native environment (e.g., the natural environment if it is naturally occurring).
  • a naturally occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or polypeptide, separated by human intervention from some or all of the co-existing materials in the natural system, is isolated.
  • Such polynucleotides could be part of a vector and/or such polynucleotides or polypeptides could be part of a composition, and still be isolated in that such vector or composition is not part of the environment in which it is found in nature.
  • CD3 cluster of differentiation 3
  • CD3 refers to 3 proteins CD3y, CD36, and CD3s that associate with the ( ⁇ -chain (zeta-chain) and TCR alpha and beta chains to form the eight protein T cell receptor (TCR) complex.
  • UniProt accession numbers P09693, P04234, P07766 provide exemplary human CD3y, CD3 6, and CD3s amino acid sequences, respectively.
  • cluster of differentiation 117 (CD117)” and “c-kit” refer to a receptor tyrosine kinase protein that, in humans, is encoded by the gene KIT, or its orthologs.
  • UniProt accession number P10721 provides exemplary human c-kit amino acid sequences.
  • NCBI Reference Sequence: NM_000222.2 provides an exemplary c-kit nucleic acid sequence.
  • an antibody molecule is a multispecific antibody molecule, e.g., it comprises a plurality of immunoglobulin variable domains sequences, wherein a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope.
  • the first and second epitopes are on the same antigen, e.g., the same protein (or subunit of a multimeric protein).
  • the first and second epitopes overlap. In an embodiment, the first and second epitopes do not overlap.
  • the first and second epitopes are on different antigens, e.g., the different proteins (or different subunits of a multimeric protein).
  • a multispecific antibody molecule comprises a third, fourth or fifth immunoglobulin variable domain.
  • a multispecific antibody molecule is a bispecific antibody molecule, a trispecific antibody molecule, or a tetraspecific antibody molecule.
  • a multispecific antibody molecule is a bispecific antibody molecule.
  • a bispecific antibody has specificity for no more than two antigens.
  • a bispecific antibody molecule is characterized by a first immunoglobulin variable domain sequence which has binding specificity for a first epitope and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope.
  • the first and second epitopes are on the same antigen, e.g., the same protein (or subunit of a multimeric protein).
  • the first and second epitopes overlap. In an embodiment, the first and second epitopes do not overlap.
  • first and second epitopes are on different antigens, e.g., the different proteins (or different subunits of a multimeric protein).
  • a bispecific antibody molecule comprises a heavy chain variable domain sequence and a light chain variable domain sequence which have binding specificity for a first epitope and 17
  • a bispecific antibody molecule comprises a half antibody having binding specificity for a first epitope and a half antibody having binding specificity for a second epitope.
  • a bispecific antibody molecule comprises a half antibody, or fragment thereof, having binding specificity for a first epitope and a half antibody, or fragment thereof, having binding specificity for a second epitope.
  • a bispecific antibody molecule comprises a scFv or a Fab, or fragment thereof, have binding specificity for a first epitope and a scFv or a Fab, or fragment thereof, have binding specificity for a second epitope.
  • an antibody molecule comprises a diabody, and a single-chain molecule, as well as an antigen-binding fragment of an antibody (e.g., Fab, F(ab’)2, and Fv).
  • an antibody molecule can include a heavy (H) chain variable domain sequence (abbreviated herein as VH), and a light (L) chain variable domain sequence (abbreviated herein as VL).
  • VH heavy chain variable domain sequence
  • VL light chain variable domain sequence
  • an antibody molecule comprises or consists of a heavy chain and a light chain (referred to herein as a half antibody.
  • an antibody molecule in another example, includes two heavy (H) chain variable domain sequences and two light (L) chain variable domain sequence, thereby forming two antigen binding sites, such as Fab, Fab’, F(ab’)2, Fc, Fd, Fd’, Fv, single chain antibodies (scFv for example), single variable domain antibodies, diabodies (Dab) (bivalent and bispecific), and chimeric (e.g., humanized) antibodies, which may be produced by the modification of whole antibodies or those synthesized de novo using recombinant DNA technologies. These functional antibody fragments retain the ability to selectively bind with their respective antigen or receptor.
  • Antibodies and antibody fragments can be from any class of antibodies including, but not limited to, IgG, IgA, IgM, IgD, and IgE, and from any subclass (e.g., IgGl, IgG2, IgG3, and IgG4) of antibodies.
  • a preparation of antibody molecules can be monoclonal or polyclonal.
  • An antibody molecule can also be a human, humanized, CDR- grafted, or in vitro generated antibody.
  • the antibody can have a heavy chain constant region chosen from, e.g., IgGl, IgG2, IgG3, or IgG4.
  • the antibody can also have a light chain chosen from, e.g., kappa or lambda.
  • immunoglobulin (Ig) is used interchangeably with the term “antibody” herein.
  • antigen-binding fragments of an antibody molecule include: (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains; (ii) a F(ab’)2 18
  • T2110-7026WO fragment a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CHI domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a diabody (dAb) fragment, which consists of a VH domain; (vi) a camelid or camelized variable domain; (vii) a single chain Fv (scFv), see e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883); (viii) a single domain antibody.
  • scFv single chain Fv
  • Antibody molecules include intact molecules as well as functional fragments thereof. Constant regions of the antibody molecules can be altered, e.g., mutated, to modify the properties of the antibody (e.g., to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, or complement function).
  • Antibody molecules can also be single domain antibodies.
  • Single domain antibodies can include antibodies whose complementary determining regions are part of a single domain polypeptide. Examples include, but are not limited to, heavy chain antibodies, antibodies naturally devoid of light chains, single domain antibodies derived from conventional 4-chain antibodies, engineered antibodies and single domain scaffolds other than those derived from antibodies.
  • Single domain antibodies may be any of the art, or any future single domain antibodies.
  • Single domain antibodies may be derived from any species including, but not limited to mouse, human, camel, llama, lamprey, fish, shark, goat, rabbit, and bovine.
  • a single domain antibody is a naturally occurring single domain antibody known as heavy chain antibody devoid of light chains. Such single domain antibodies are disclosed in WO 9404678, for example.
  • variable domain derived from a heavy chain antibody naturally devoid of light chain is known herein as a VHH or nanobody to distinguish it from the conventional VH of four chain immunoglobulins.
  • VHH molecule can be derived from antibodies raised in Camelidae species, for example in camel, llama, dromedary, alpaca and guanaco. Other species besides Camelidae may produce heavy chain antibodies naturally devoid of light chain; such VHHs are within the scope of the invention.
  • VH and VL regions can be subdivided into regions of hypervariability, termed “complementarity determining regions” (CDR), interspersed with regions that are more conserved, termed “framework regions” (FR or FW).
  • CDR complementarity determining regions
  • FR framework regions
  • CDR complementarity determining region
  • the precise amino acid sequence boundaries of a given CDR can be determined using any of a number of known schemes, including those described by Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (“Kabat” numbering scheme), Al-Lazikani et al., (1997) JMB 273,927-948 (“Chothia” numbering scheme). As used herein, the CDRs defined according the “Chothia” number scheme are also sometimes referred to as “hypervariable loops.”
  • the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3).
  • the CDR amino acids in the VH are numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3); and the amino acid residues in VL are numbered 26-32 (LCDR1), 50-52 (LCDR2), and 91-96 (LCDR3).
  • Each VH and VL typically includes three CDRs and four FRs, arranged from aminoterminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
  • the antibody molecule can be a polyclonal or a monoclonal antibody.
  • monoclonal antibody or “monoclonal antibody composition” as used herein refer to a preparation of antibody molecules of single molecular composition.
  • a monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope.
  • a monoclonal antibody can be made by hybridoma technology or by methods that do not use hybridoma technology (e.g., recombinant methods).
  • the antibody can be recombinantly produced, e.g., produced by phage display or by combinatorial methods.
  • Phage display and combinatorial methods for generating antibodies are known in the art (as described in, e.g., Ladner et al. U.S. Patent No. 5,223,409; Kang et al. International Publication No. WO 92/18619; Dower et al. International Publication No. WO 91/17271; Winter et al. International Publication WO 92/20791; Markland et al. International Publication No. WO 92/15679; Breitling et al. International Publication WO 93/01288; McCafferty et al. International Publication No. WO 92/01047; Garrard et al. International Publication No.
  • the antibody is a fully human antibody (e.g., an antibody made in a mouse which has been genetically engineered to produce an antibody from a human immunoglobulin sequence), or a non-human antibody, e.g., a rodent (mouse or rat), goat, primate (e.g., monkey), camel antibody.
  • a rodent mouse or rat
  • the non-human antibody is a rodent (mouse or rat antibody).
  • Methods of producing rodent antibodies are known in the art.
  • Human monoclonal antibodies can be generated using transgenic mice carrying the human immunoglobulin genes rather than the mouse system. Splenocytes from these transgenic mice immunized with the antigen of interest are used to produce hybridomas that secrete human mAbs with specific affinities for epitopes from a human protein (see, e.g., Wood et al. International Application WO 91/00906, Kucherlapati et al. PCT publication WO 91/10741; Lonberg et al. International Application WO 92/03918; Kay et al. International Application 21
  • An antibody molecule can be one in which the variable region, or a portion thereof, e.g., the CDRs, are generated in a non-human organism, e.g., a rat or mouse. Chimeric, CDR-grafted, and humanized antibodies are within the invention. Antibody molecules generated in a non- human organism, e.g., a rat or mouse, and then modified, e.g., in the variable framework or constant region, to decrease antigenicity in a human are within the invention.
  • An “effectively human” protein is a protein that does substantially not evoke a neutralizing antibody response, e.g., the human anti-murine antibody (HAMA) response.
  • HAMA can be problematic in a number of circumstances, e.g., if the antibody molecule is administered repeatedly, e.g., in treatment of a chronic or recurrent disease condition.
  • a HAMA response can make repeated antibody administration potentially ineffective because of an increased antibody clearance from the serum (see, e.g., Saleh et al. Cancer Immunol. Immunother., 32:180-190 (1990)) and also because of potential allergic reactions (see, e.g., LoBuglio et al., Hybridoma, 5:5117-5123 (1986)).
  • Chimeric antibodies can be produced by recombinant DNA techniques known in the art (see Robinson et al., International Patent Publication PCT/US86/02269; Akira, et al., European Patent Application 184,187; Taniguchi, M., European Patent Application 171,496; Morrison et al., European Patent Application 173,494; Neuberger et al., International Application WO 86/01533; Cabilly et al. U.S. Patent No. 4,816,567; Cabilly et al., European Patent Application 125,023; Better et al. (1988 Science 240:1041-1043); Liu et al.
  • a humanized or CDR-grafted antibody will have at least one or two but generally all three recipient CDRs (of heavy and or light immunoglobulin chains) replaced with a donor CDR.
  • the antibody may be replaced with at least a portion of a non-human CDR or only some of the CDRs may be replaced with non-human CDRs. It is only necessary to replace the number of CDRs required for binding to the antigen.
  • the donor will be a rodent antibody, e.g., a 22
  • the recipient will be a human framework or a human consensus framework.
  • the immunoglobulin providing the CDRs is called the "donor” and the immunoglobulin providing the framework is called the "acceptor.”
  • the donor immunoglobulin is a non-human (e.g., rodent).
  • the acceptor framework is a naturally occurring e.g., a human) framework or a consensus framework, or a sequence about 85% or higher, preferably 90%, 95%, 99% or higher identical thereto.
  • Consensus sequence refers to the sequence formed from the most frequently occurring amino acids (or nucleotides) in a family of related sequences (See e.g., Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987). In a family of proteins, each position in the consensus sequence is occupied by the amino acid occurring most frequently at that position in the family. If two amino acids occur equally frequently, either can be included in the consensus sequence.
  • a “consensus framework” refers to the framework region in the consensus immunoglobulin sequence.
  • An antibody molecule can be humanized by methods known in the art (see e.g., Morrison, S. L., 1985, Science 229:1202-1207, by Oi et al., 1986, BioTechniques 4:214, and by Queen et al. US 5,585,089, US 5,693,761 and US 5,693,762, the contents of all of which are hereby incorporated by reference).
  • Humanized or CDR-grafted antibody molecules can be produced by CDR-grafting or CDR substitution, wherein one, two, or all CDRs of an immunoglobulin chain can be replaced.
  • CDR-grafting or CDR substitution wherein one, two, or all CDRs of an immunoglobulin chain can be replaced.
  • humanized antibody molecules in which specific amino acids have been substituted, deleted or added. Criteria for selecting amino acids from the donor are described in US 5,585,089, e.g., columns 12-16 of US 5,585,089, e.g., columns 12-16 of US 5,585,089, the contents of which are hereby incorporated by reference. Other techniques for humanizing antibodies are described in Padlan et al. EP 519596 Al, published on December 23, 1992.
  • the antibody molecule can be a single chain antibody.
  • a single-chain antibody (scFV) may be engineered (see, for example, Colcher, D. et al. (1999) Ann N Y Acad Sci 880:263-80; and Reiter, Y. (1996) Clin Cancer Res 2:245-52).
  • the single chain antibody can be dimerized or multimerized to generate multivalent antibodies having specificities for different epitopes of the same target protein.
  • the antibody molecule has a heavy chain constant region chosen from, e.g., the heavy chain constant regions of IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE; particularly, chosen from, e.g., the (e.g., human) heavy chain constant regions of IgGl, IgG2, IgG3, and IgG4.
  • the antibody molecule has a light chain constant region chosen from, e.g., the (e.g., human) light chain constant regions of kappa or lambda.
  • the constant region can be altered, e.g., mutated, to modify the properties of the antibody (e.g., to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, and/or complement function).
  • the antibody has: effector function; and can fix complement.
  • the antibody does not; recruit effector cells; or fix complement.
  • the antibody has reduced or no ability to bind an Fc receptor. For example, it is an isotype or subtype, fragment or other mutant, which does not support binding to an Fc receptor, e.g., it has a mutagenized or deleted Fc receptor binding region.
  • Antibodies with altered function e.g. altered affinity for an effector ligand, such as FcR on a cell, or the Cl component of complement can be produced by replacing at least one amino acid residue in the constant portion of the antibody with a different residue (see e.g., EP 388,151 Al, U.S. Pat. No. 5,624,821 and U.S. Pat. No. 5,648,260, the contents of all of which are hereby incorporated by reference). Similar type of alterations could be described which if applied to the murine, or other species immunoglobulin would reduce or eliminate these functions.
  • the multispecific antibody molecule comprises one or more mutations that reduce and/or eliminate binding of the antibody molecule to FcyRI, II, and/or III.
  • Exemplary mutations include one or more of Eeu234 (e.g., Eeu234Ala), Eeu235 (e.g., Eeu235Ala) (e.g., EAEA mutations), or Pro329 (e.g., Pro329Gly) (e.g., EAEA-PG mutations).
  • An antibody molecule can be derivatized or linked to another functional molecule (e.g., another peptide or protein).
  • a "derivatized" antibody molecule is one that has 24
  • Methods of derivatization include but are not limited to the addition of a fluorescent moiety, a radionucleotide, a toxin, an enzyme or an affinity ligand such as biotin. Accordingly, the antibody molecules of the invention are intended to include derivatized and otherwise modified forms of the antibodies described herein, including immunoadhesion molecules.
  • an antibody molecule can be functionally linked (by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or a diabody), a detectable agent, a cytotoxic agent, a pharmaceutical agent, and/or a protein or peptide that can mediate association of the antibody or antibody portion with another molecule (such as a streptavidin core region or a poly histidine tag).
  • another antibody e.g., a bispecific antibody or a diabody
  • detectable agent e.g., a detectable agent, a cytotoxic agent, a pharmaceutical agent, and/or a protein or peptide that can mediate association of the antibody or antibody portion with another molecule (such as a streptavidin core region or a poly histidine tag).
  • One type of derivatized antibody molecule is produced by crosslinking two or more antibodies (of the same type or of different types, e.g., to create bispecific antibodies).
  • Suitable crosslinkers include those that are heterobifunctional, having two distinctly reactive groups separated by an appropriate spacer (e.g., m-maleimidobenzoyl-N-hydroxysuccinimide ester) or homobifunctional (e.g., disuccinimidyl suberate).
  • Such linkers are available from Pierce Chemical Company, Rockford, Ill.
  • Exemplary structures of multispecific antibody molecules disclosed herein are described throughout. Exemplary structures are further described, e.g., in: Weidle U et al. (2013) The Intriguing Options of Multispecific Antibody Formats for Treatment of Cancer. Cancer Genomics & Proteomics 10: 1-18 (2013); and Spiess C et al. (2015) Alternative molecular formats and therapeutic applications for bispecific antibodies. Molecular Immunology 67: 95- 106; the full contents of each of which is incorporated by reference herein).
  • multispecific antibody molecules can comprise more than one antigen-binding site, where different sites are specific for different antigens. In some embodiments, multispecific antibody molecules can bind more than one (e.g., two or more) epitopes on the same antigen. In some embodiments, multispecific antibody molecules comprise an antigen-binding site specific for a target cell (e.g., cancer cell) and a different antigen-binding site specific for an immune effector cell. In one embodiment, the multispecific antibody molecule is a bispecific antibody molecule. Bispecific antibody molecules can be classified into 25
  • BsIgG is a format that is monovalent for each antigen.
  • Exemplary BsIgG formats include but are not limited to crossMab, DAF (two-in-one), DAF (four- in-one), DutaMab, DT-IgG, knobs-in-holes common EC, knobs-in-holes assembly, charge pair, Fab-arm exchange, SEEDbody, triomab, LUZ-Y, Fcab, Kk-body, orthogonal Fab. See Spiess et al. Mol. Immunol. 67(2015):95-106.
  • BsIgGs include catumaxomab (Fresenius Biotech, Trion Pharma, Neopharm), which contains an anti-CD3 arm and an anti-EpCAM arm; and ertumaxomab (Neovii Biotech, Fresenius Biotech), which targets CD3 and HER2.
  • BsIgG comprises heavy chains that are engineered for heterodimerization.
  • heavy chains can be engineered for heterodimerization using a “knobs-into-holes” strategy, a SEED platform, a common heavy chain (e.g., in Kk-bodies), and use of heterodimeric Fc regions. See Spiess et al. Mol. Immunol.
  • BsIgG can be produced by separate expression of the component antibodies in different host cells and subsequent purification/assembly into a BsIgG.
  • BsIgG can also be produced by expression of the component antibodies in a single host cell.
  • BsIgG can be purified using affinity chromatography, e.g., using protein A and sequential pH elution.
  • IgG appended with an additional antigen-binding moiety is another format of bispecific antibody molecules.
  • monospecific IgG can be engineered to have bispecificity by appending an additional antigen-binding unit onto the monospecific IgG, e.g., at the N- or C- terminus of either the heavy or light chain.
  • additional antigen-binding units include single domain antibodies (e.g., variable heavy chain or variable light chain), engineered protein scaffolds, and paired antibody variable domains (e.g., single chain variable fragments or variable fragments). See Id.
  • Examples of appended IgG formats include dual variable domain IgG (DVD-Ig), IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)IgG, IgG(L,H)-Fv, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, zybody, and D VI- IgG (four- in-one). See Spiess et al. Mol. Immunol. 67(2015):95-106.
  • An example of an IgG-scFv is MM- 141 (Merrimack Pharmaceuticals), which binds IGF-1R and HER3. Examples 26
  • T2110-7026WO of DVD-Ig include ABT-981 (AbbVie), which binds IL-la and IL-1P; and ABT-122 (AbbVie), which binds TNF and IL-17A.
  • Bispecific antibody fragments are a format of bispecific antibody molecules that lack some or all of the antibody constant domains. For example, some BsAb lack an Fc region.
  • bispecific antibody fragments include heavy and light chain regions that are connected by a peptide linker that permits efficient expression of the BsAb in a single host cell.
  • Exemplary bispecific antibody fragments include but are not limited to nanobody, nanobody-HAS, BiTE, Diabody, DART, TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, triple body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL- scFv, F(ab’)2, F(ab’)2-scFv2, scFv-KIH, Fab-scFv-Fc, tetravalent HCAb, scDiabody-Fc, Diabody-Fc, tandem scFv-Fc, and intrabody. See Id.
  • the BiTE format comprises tandem scFvs, where the component scFvs bind to CD3 on T cells and a surface antigen on cancer cells.
  • the antibody molecule may include a 1+1 or a 2+1 mixed valency configuration enabled by a heterodimeric Fc, e.g., a 1+1 Fab-scFv-Fc format, e.g., as described in Moore, et al. (2019) Methods 154:38-50, the contents of which are incorporated by reference herein in its entirety, including the passages describing various heterodimeric antibody configurations.
  • Bispecific fusion proteins include antibody fragments linked to other proteins, e.g., to add additional specificity and/or functionality.
  • An example of a bispecific fusion protein is an immTAC, which comprises an anti-CD3 scFv linked to an affinity-matured T-cell receptor that recognizes HLA-presented peptides.
  • the dock-and-lock (DNL) method can be used to generate bispecific antibody molecules with higher valency.
  • fusions to albumin binding proteins or human serum albumin can be extend the serum half-life of antibody fragments. See Id.
  • chemical conjugation e.g., chemical conjugation of antibodies and/or antibody fragments
  • An exemplary bispecific antibody conjugate includes the CovX-body format, in which a low molecular weight drug is conjugated site-specifically to a single reactive lysine in each Fab arm or an antibody or fragment thereof.
  • the conjugation improves the serum half-life of the low molecular weight drug.
  • An exemplary CovX-body is CVX-241 (NCT01004822), which comprises an antibody conjugated to two short peptides inhibiting either VEGF or Ang2. See Id. 27 4374195.1 Attorney Docket No.: T2110-7026WO
  • the antibody molecules can be produced by recombinant expression, e.g., of at least one or more component, in a host system.
  • host systems include eukaryotic cells (e.g., mammalian cells, e.g., CHO cells, or insect cells, e.g., SF9 or S2 cells) and prokaryotic cells (e.g., E. coll).
  • eukaryotic cells e.g., mammalian cells, e.g., CHO cells, or insect cells, e.g., SF9 or S2 cells
  • prokaryotic cells e.g., E. coll
  • the selection of the appropriate host cell may be based on several considerations, e.g., the capability of the host cell to have certain post-translational modifications (e.g., glycosylation).
  • E. coli host cells may be utilized to produce aglycosylated multispecific antibody molecules.
  • Bispecific antibody molecules can be produced by separate expression of the components in different host cells and subsequent purification/assembly. Alternatively, the antibody molecules can be produced by expression of the components in a single host cell. Purification of bispecific antibody molecules can be performed by various methods such as affinity chromatography, e.g., using protein A and sequential pH elution. In other embodiments, affinity tags can be used for purification, e.g., histidine-containing tag, myc tag, or streptavidin tag.
  • the antibody molecule is a CDR-grafted scaffold domain.
  • the scaffold domain is based on a fibronectin domain, e.g., fibronectin type III domain.
  • the overall fold of the fibronectin type III (Fn3) domain is closely related to that of the smallest functional antibody fragment, the variable domain of the antibody heavy chain. There are three loops at the end of Fn3; the positions of BC, DE and FG loops approximately correspond to those of CDR1, 2 and 3 of the VH domain of an antibody.
  • Fn3 does not have disulfide bonds; and therefore, Fn3 is stable under reducing conditions, unlike antibodies and their fragments (see, e.g., WO 98/56915; WO 01/64942; WO 00/34784).
  • An Fn3 domain can be modified (e.g., using CDRs or hypervariable loops described herein) or varied, e.g., to select domains that bind to an antigen/marker/cell described herein.
  • a scaffold domain e.g., a folded domain
  • an antibody e.g., a “minibody” scaffold created by deleting three beta strands from a heavy chain variable domain of a monoclonal antibody (see, e.g., Tramontane et al., 1994, J Mol. Recognit. 7:9; and Martin et al., 1994, EMBO J. 13:5303-5309).
  • the “minibody” can be used to present two hypervariable loops.
  • the scaffold domain is a V-like domain (see, e.g., Coia et al. WO 99/45110) or a domain derived from tendamistatin, which is a 74 residue, six-
  • T2110-7026WO strand beta sheet sandwich held together by two disulfide bonds see, e.g., McConnell and Hoess, 1995, J Mol. Biol. 250:460.
  • the loops of tendamistatin can be modified (e.g., using CDRs or hypervariable loops) or varied, e.g., to select domains that bind to a marker/antigen/cell described herein.
  • Another exemplary scaffold domain is a beta-sandwich structure derived from the extracellular domain of CTLA-4 (see, e.g., WO 00/60070).
  • exemplary scaffold domains include but are not limited to T-cell receptors; MHC proteins; extracellular domains (e.g., fibronectin Type III repeats, EGF repeats); protease inhibitors (e.g., Kunitz domains, ecotin, BPTI, and so forth); TPR repeats; trifoil structures; zinc finger domains; DNA-binding proteins; particularly monomeric DNA binding proteins; RNA binding proteins; enzymes, e.g., proteases (particularly inactivated proteases), RNase; chaperones, e.g., thioredoxin, and heat shock proteins; and intracellular signaling domains (such as SH2 and SH3 domains). See, e.g., US 20040009530 and US 7,501,121, incorporated herein by reference.
  • extracellular domains e.g., fibronectin Type III repeats, EGF repeats
  • protease inhibitors e.g., Kunitz domains, ecotin, BPTI, and so
  • a scaffold domain is evaluated and chosen, e.g., by one or more of the following criteria: (1) amino acid sequence, (2) sequences of several homologous domains, (3) 3-dimensional structure, and/or (4) stability data over a range of pH, temperature, salinity, organic solvent, oxidant concentration.
  • the scaffold domain is a small, stable protein domain, e.g., a protein of less than 100, 70, 50, 40 or 30 amino acids.
  • the domain may include one or more disulfide bonds or may chelate a metal, e.g., zinc.
  • a variety of formats can be generated which contain additional binding entities attached to the N or C terminus of antibodies. These fusions with single chain or disulfide stabilized Fvs or Fabs result in the generation of tetravalent molecules with bivalent binding specificity for each antigen. Combinations of scFvs and scFabs with IgGs enable the production of molecules which can recognize three or more different antigens.
  • Antibody-Fab fusions are bispecific antibodies comprising a traditional antibody to a first target and a Fab to a second target fused to the C terminus of the antibody heavy chain. Commonly the antibody and the Fab will have a common light chain. Antibody fusions can be 29
  • Antibody-scFv fusions are bispecific antibodies comprising a traditional antibody and a scFv of unique specificity fused to the C terminus of the antibody heavy chain.
  • the scFv can be fused to the C terminus through the Heavy Chain of the scFv either directly or through a linker peptide.
  • Antibody fusions can be produced by (7 engineering the DNA sequence of the target fusion, and (2) transfecting the target DNA into a suitable host cell to express the fusion protein. It seems like the antibody-scFv fusion may be linked by a (Gly)-Ser linker between the C- terminus of the CH3 domain and the N-terminus of the scFv, as described by Coloma, J. et al. (1997) Nature Biotech 15:159.
  • VD dual variable domain immunoglobulin
  • exemplary multispecific antibody formats include, e.g., those described in the following US20160114057A1, US20130243775A1, US20140051833, US20130022601, US20150017187A1, US20120201746A1, US20150133638A1, US20130266568A1, US20160145340A1, WO2015127158A1, US20150203591A1, US20140322221A1, US20130303396A1, US20110293613, US20130017200A1, US20160102135A1, WO2015197598A2, WO2015197582A1, US9359437, US20150018529, WO2016115274 Al, WO201 6087416A1, US20080069820A1, US9145588B, US7919257, and US20150232560A1.
  • Exemplary multispecific antibody molecules utilizing a full antibody-Fab/scFab format include those described in the following, US9382323B2, US20140072581A1, US20140308285A1, US20130165638A1, US20130267686A1, US20140377269A1, US7741446B2, and WO1995009917A1.
  • Exemplary multispecific antibody molecules utilizing a domain exchange format include those described in the following, US20150315296A1, W02016087650A1,
  • Fc-containing entities also known as mini-antibodies
  • Fc-containing entities can be generated by fusing scFv to the C-termini of constant heavy region domain 3 (CH3-scFv) and/or to the hinge region (scFv- hinge-Fc) of an antibody with a different specificity.
  • Trivalent entities can also be made which have disulfide stabilized variable domains (without peptide linker) fused to the C-terminus of CH3 domains of IgGs.
  • the multispecific antibody molecules disclosed herein includes an immunoglobulin constant region (e.g., an Fc region).
  • Fc regions can be chosen from the heavy chain constant regions of IgGl, IgG2, IgG3 or IgG4; more particularly, the heavy chain constant region of human IgGl, IgG2, IgG3, or IgG4.
  • the immunoglobulin chain constant region (e.g., the Fc region) is altered, e.g., mutated, to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, or complement function.
  • an interface of a first and second immunoglobulin chain constant regions is altered, e.g., mutated, to increase or decrease dimerization, e.g., relative to a non-engineered interface, e.g., a naturally occurring interface.
  • dimerization of the immunoglobulin chain constant region can be enhanced by providing an Fc interface of a first and a second Fc region with one or more of: a paired protuberance-cavity (“knob-in-a hole”), an electrostatic interaction, or a strand-exchange, such that a greater ratio of heteromultimer to homomultimer forms, e.g., relative to a nonengineered interface.
  • the multispecific antibody molecules include a paired amino acid substitution at a position chosen from one or more of 347, 349, 350, 351, 366, 368, 370, 392, 394, 395, 397, 398, 399, 405, 407, or 409, e.g., of the Fc region of human IgGl
  • the immunoglobulin chain constant region e.g., Fc region
  • T2110-7026WO substitution chosen from: T366S, L368A, or Y407V (e.g., corresponding to a cavity or hole), and T366W (e.g., corresponding to a protuberance or knob).
  • the molecule includes a half-life extender, e.g., a human serum albumin or an antibody molecule to human serum albumin.
  • a half-life extender e.g., a human serum albumin or an antibody molecule to human serum albumin.
  • multispecific antibody formats and methods of making said multispecific antibodies are also disclosed in e.g., Speiss et al. Molecular Immunology 67 (2015) 95-106; and Klein et al mAbs 4:6, 653-663; November/December 2012; the entire contents of each of which are incorporated by reference herein.
  • Heterodimerized bispecific antibodies are based on the natural IgG structure, wherein the two binding arms recognize different antigens.
  • IgG derived formats that enable defined monovalent (and simultaneous) antigen binding are generated by forced heavy chain heterodimerization, combined with technologies that minimize light chain mispairing (e.g., common light chain). Forced heavy chain heterodimerization can be obtained using, e.g., knob- in-hole OR strand exchange engineered domains (SEED).
  • Knob-in-Hole as described in US 5,731,116, US 7,476,724 and Ridgway, J. et al. (1996) Prot. Engineering 9(7): 617-621, broadly involves: (1) mutating the CH3 domain of one or both antibodies to promote heterodimerization; and (2) combining the mutated antibodies under conditions that promote heterodimerization.
  • “Knobs” or “protuberances” are typically created by replacing a small amino acid in a parental antibody with a larger amino acid (e.g., T366Y or T366W); “Holes” or “cavities” are created by replacing a larger residue in a parental antibody with a smaller amino acid (e.g., Y407T, T366S, L368A and/or Y407V).
  • Exemplary KiH mutations include S354C, T366W in the “knob” heavy chain and Y349C, T366S, L368A, Y407V in the “hole” heavy chain.
  • Other exemplary KiH mutations are provided in Table 1, with additional optional stabilizing Fc cysteine mutations.
  • Fc mutations are provided by Igawa and Tsunoda who identified 3 negatively charged residues in the CH3 domain of one chain that pair with three positively charged residues in the CH3 domain of the other chain. These specific charged residue pairs are: E356-K439, E357-K370, D399-K409 and vice versa.
  • E356K, E357K and D399K as well as K370E, K409D, K439E in chain B, alone or in combination with newly identified disulfide bridges, they were able to favor very efficient heterodimerization while suppressing homodimerization at the same time (Martens T et al.
  • a novel one-armed antic- Met antibody inhibits glioblastoma growth in vivo. Clin Cancer Res 2006; 12:6144-52; PMID: 17062691).
  • Xencor defined 41 variant pairs based on combining structural calculations and sequence information that were subsequently screened for maximal heterodimerization, defining the combination of S364H, F405A (HA) on chain A and Y349T, T394F on chain B (TF) (Moore GE et al.
  • a novel bispecific antibody format enables
  • exemplary Fc mutations to promote heterodimerization of multispecific antibodies include those described in the following references, the contents of each of which is incorporated by reference herein, WO2016071377A1, US20140079689A1, US20160194389A1, US20160257763, WO2016071376A2, WO2015107026 Al, WO2015107025 Al, W02015107015A1, US20150353636A1, US20140199294A1, US7750128B2, US20160229915A1, US20150344570A1, US8003774A1, US20150337049A1, US20150175707A1, US20140242075A1, US20130195849A1, US20120149876A1, US20140200331A1, US9309311B2, US8586713, US20140037621A1, US20130178605A1, US20140363426A1, US20140051835A1 and US20110054151A1.
  • Stabilizing cysteine mutations have also been used in combination with KiH and other Fc heterodimerization promoting variants, see e.g., US7183076.
  • Other exemplary cysteine modifications include, e.g., those disclosed in US20140348839A1, US7855275B2, and US9000130B2.
  • Heterodimeric Fc platform that support the design of bispecific and asymmetric fusion proteins by devising strand-exchange engineered domain (SEED) C(H)3 heterodimers are known.
  • SEED strand-exchange engineered domain
  • These derivatives of human IgG and IgA C(H)3 domains create complementary human SEED C(H)3 heterodimers that are composed of alternating segments of human IgA and IgG C(H)3 sequences.
  • the resulting pair of SEED C(H)3 domains preferentially associates to form heterodimers when expressed in mammalian cells.
  • SEEDbody (Sb) fusion proteins consist of [IgGl hinge] -C(H)2- [SEED C(H)3], that may be genetically linked to one or more fusion partners (see e.g., Davis JH et al. SEEDbodies: fusion proteins based on strand exchange engineered domain (SEED) CH3 heterodimers in an Fc analogue platform for asymmetric binders or immunofusions and bispecific antibodies. Protein Eng Des Sei 2010; 23:195-202; PMID:20299542 and US8871912. The contents of each of which are incorporated by reference herein).
  • Duobody technology to produce bispecific antibodies with correct heavy chain pairing are known.
  • the DuoBody technology involves three basic steps to generate stable bispecific human IgGl antibodies in a post-production exchange reaction. In a first step, two IgGls, each containing single matched mutations in the third constant (CH3) domain, are produced separately using standard mammalian recombinant cell lines. Subsequently, these IgGl antibodies are purified according to standard processes for recovery and purification.
  • Light chain mispairing needs to be avoided to generate homogenous preparations of bispecific IgGs.
  • One way to achieve this is through the use of the common light chain principle, i.e. combining two binders that share one light chain but still have separate specificities.
  • An exemplary method of enhancing the formation of a desired bispecific antibody from a mixture of monomers is by providing a common variable light chain to interact with each of the heteromeric 35
  • CrossMab technology Another option to reduce light chain mispairing is the CrossMab technology which avoids non-specific L chain mispairing by exchanging CHI and CL domains in the Fab of one half of the bispecific antibody. Such crossover variants retain binding specificity and affinity, but make the two arms so different that L chain mispairing is prevented.
  • the CrossMab technology (as reviewed in Klein et al. Supra) involves domain swapping between heavy and light chains so as to promote the formation of the correct pairings. Briefly, to construct a bispecific IgG-like CrossMab antibody that could bind to two antigens by using two distinct light chain-heavy chain pairs, a two-step modification process is applied.
  • a dimerization interface is engineered into the C-terminus of each heavy chain using a heterodimerization approach, e.g., Knob-into-hole (KiH) technology, to ensure that only a heterodimer of two distinct heavy chains from one antibody (e.g., Antibody A) and a second antibody (e.g., Antibody B) is efficiently formed.
  • a heterodimerization approach e.g., Knob-into-hole (KiH) technology
  • CHI constant heavy 1
  • An exemplary method of enhancing the formation of a desired bispecific antibody from a mixture of monomers is by providing a common variable heavy chain to interact with each of the heteromeric variable light chain regions of the bispecific antibody.
  • Compositions and methods of producing bispecific antibodies with a common heavy chain are disclosed in, e.g., 36
  • compositions and methods of producing multispecific antibodies with correct light chain pairing include various amino acid modifications.
  • Zymeworks describes heterodimers with one or more amino acid modifications in the CHI and/or CL domains, one or more amino acid modifications in the VH and/or VL domains, or a combination thereof, which are part of the interface between the light chain and heavy chain and create preferential pairing between each heavy chain and a desired light chain such that when the two heavy chains and two light chains of the heterodimer pair are co-expressed in a cell, the heavy chain of the first heterodimer preferentially pairs with one of the light chains rather than the other (see e.g., W02015181805).
  • Other exemplary methods are described in WO2016026943 (Argen-X), US20150211001, US20140072581A1, US20160039947A1, and US20150368352.
  • Multispecific antibody molecules may also include amino acid modifications that affect the glycosylation of the molecule. For example, these modifications may serve to increase or decrease the extent to which the molecule is glycosylated. These additions or deletions of glycosylation sites may additionally alter the effector function of the multispecific antibody molecule (e.g., reduce antibodydependent cellular cytotoxicity (ADCC)-mediated effector function).
  • ADCC antibodydependent cellular cytotoxicity
  • IgG antibody molecules contain a conserved glycosylation site at amino acid N297 (EU numbering) in the CH2 domain.
  • This conserved glycan contains A-acctylglucosaminc (GlcNAc) and mannose at its core and additionally may comprise bisecting GlcNAc, fucose, galactose, and sialic acid.
  • This glycosylation mediates antibody interactions with FcyRs and Clq to facilitate ADCC.
  • the multispecific antibody molecules described herein may comprise an aglycosylation mutation (e.g., a substitution mutation) at position N297 (EU numbering).
  • exemplary aglycosylation substitution mutations include N297G, N297A, and N297Q.
  • Multispecific antibody molecules may also include amino acid modifications that affect the ability of the molecule to undergo FcRn recycling.
  • Binding of an IgG antibody molecule to the MHC class I-related receptor, FcRn recycles the endocytosed antibody molecule from the endosome back to the bloodstream, rescuing the antibody molecule from intracellular degradation and extending antibody serum half-life.
  • FcRn interaction with the Fc portion of IgG occurs at the CH2 and CH3 domain interface and involves the Fc amino acid residues at positions 252-256 in the CH2 domains and at 310, 433, 434, and 435 in the CH3 domains.
  • the multispecific antibody molecules described herein may comprise a substitution mutation to reduce the ability of the molecule to bind to and undergo FcRn recycling (e.g., to reduce the antibody half-life).
  • exemplary substitution mutations include alanine substitution mutations at positions Ile253, Ser254, His435, Tyr436, Glu233-Gly236, Arg255, Lys288, Ser415, His310, and His433.
  • Multispecific antibody molecules e.g., multispecific antibody molecules
  • multispecific antibody molecules that include the lambda light chain polypeptide and a kappa light chain polypeptides
  • Methods for generating bispecific antibody molecules comprising the lambda light chain polypeptide and a kappa light chain polypeptides are disclosed in PCT/US 17/53053 filed on September 22, 2017, incorporated herein by reference in its entirety.
  • the multispecific antibody molecules include a multispecific antibody molecule, e.g., an antibody molecule comprising two binding specificities, e.g., a bispecific antibody molecule.
  • the multispecific antibody molecule includes: a lambda light chain polypeptide 1 (LLCP1) specific for a first epitope; a heavy chain polypeptide 1 (HCP1) specific for the first epitope; a kappa light chain polypeptide 2 (KLCP2) specific for a second epitope; and a heavy chain polypeptide 2 (HCP2) specific for the second epitope.
  • LLCP1 lambda light chain polypeptide 1
  • HCP1 heavy chain polypeptide 1
  • KLCP2 kappa light chain polypeptide 2
  • HCP2 heavy chain polypeptide 2
  • LLCP1 Lambda light chain polypeptide 1
  • LC light chain
  • an LLCP1 comprises LC-CDR1, LC-CDR2, LC-CDR3, FR1, FR2, FR3, FR4, and CHI, or sufficient sequence therefrom to mediate specific binding of its epitope and complex with an HCP1. LLCP1, together with its HCP1, provide specificity for a first epitope (while KLCP2, together with its HCP2, provide specificity for a second epitope). As described elsewhere herein, LLCP1 has a higher affinity for HCP1 than for HCP2.
  • KLCP2 Kappa light chain polypeptide 2
  • LC sufficient light chain
  • a KLCP2 comprises LC-CDR1, LC-CDR2, LC-CDR3, FR1, FR2, FR3, FR4, and CHI, or sufficient sequence therefrom to mediate specific binding of its epitope and complex with an HCP2.
  • KLCP2, together with its HCP2 provide specificity for a second epitope (while LLCP1, together with its HCP1, provide specificity for a first epitope).
  • Heavy chain polypeptide 1 refers to a polypeptide comprising sufficient heavy chain (HC) sequence, e.g., HC variable region sequence, such that when combined with a cognate LLCP1, can mediate specific binding to its epitope and complex with an HCP1.
  • HC sufficient heavy chain
  • it comprises all or a fragment of a CHlregion.
  • it comprises all or a fragment of a CH2 and/or CH3 region.
  • an HCP1 comprises HC-CDR1, HC-CDR2, HC-CDR3, FR1, FR2, FR3, FR4, CHI, CH2, and CH3, or sufficient sequence therefrom to: (i) mediate specific binding of its epitope and complex with an LLCP1, (ii) to complex preferentially, as described herein to LLCP1 as opposed to KLCP2; and (iii) to complex preferentially, as described herein, to an HCP2, as opposed to another molecule of HCP1.
  • HCP1, together with its LLCP1 provide specificity for a first epitope (while KLCP2, together with its HCP2, provide specificity for a second epitope).
  • Heavy chain polypeptide 2 refers to a polypeptide comprising sufficient heavy chain (HC) sequence, e.g., HC variable region sequence, such that when combined with a cognate LLCP1, can mediate specific binding to its epitope and complex with an HCP1.
  • HC sufficient heavy chain
  • it comprises all or a fragment of a CHlregion.
  • it comprises all or a fragment of a CH2 and/or CH3 region.
  • an HCP1 comprises HC-CDR1, HC-CDR2, HC-CDR3, FR1, FR2, FR3, FR4, CHI, CH2, and CH3, 39
  • LLCP1 has a higher affinity for HCP1 than for HCP2; and/or KLCP2 has a higher affinity for HCP2 than for HCP1.
  • the affinity of LLCP1 for HCP1 is sufficiently greater than its affinity for HCP2, such that under preselected conditions, e.g., in aqueous buffer, e.g., at pH 7, in saline, e.g., at pH 7, or under physiological conditions, at least 75%, 80, 90, 95, 98, 99, 99.5, or 99.9 % of the multispecific antibody molecule molecules have a LLCP1 complexed, or interfaced with, a HCPl.
  • the HCP1 has a greater affinity for HCP2, than for a second molecule of HCP1; and/or the HCP2 has a greater affinity for HCP1, than for a second molecule of HCP2.
  • the affinity of HCP1 for HCP2 is sufficiently greater than its affinity for a second molecule of HCP1, such that under preselected conditions, e.g., in aqueous buffer, e.g., at pH 7, in saline, e.g., at pH 7, or under physiological conditions, at least 75%, 80, 90, 95, 98, 99 99.5 or 99.9 % of the multispecific antibody molecule molecules have a HCP1 complexed, or interfaced with, a HCP2.
  • a method for making, or producing, a multispecific antibody molecule e.g., as a first or second tumor-targeting moiety of a multispecific antibody molecule polypeptide of the invention.
  • the method includes:
  • a first heavy chain polypeptide e.g., a heavy chain polypeptide comprising one, two, three or all of a first heavy chain variable region (first VH), a first CHI, a first heavy chain constant region (e.g., a first CH2, a first CH3, or both)
  • first VH first heavy chain variable region
  • first CHI first heavy chain constant region
  • first CH2 first CH2, a first CH3, or both
  • a second heavy chain polypeptide e.g., a heavy chain polypeptide comprising one, two, three or all of a second heavy chain variable region (second VH), a second CHI, a second heavy chain constant region (e.g., a second CH2, a second CH3, or both)
  • second VH second heavy chain variable region
  • second CHI second heavy chain variable region
  • second CH2 second CH3, or both
  • a lambda chain polypeptide e.g., a lambda light variable region (VLk), a lambda light constant chain (VLk), or both
  • VLk lambda light variable region
  • VLk lambda light constant chain
  • a kappa chain polypeptide e.g., a kappa light variable region (VLK), a kappa light constant chain (VLK), or both
  • VLK kappa light variable region
  • VLK kappa light constant chain
  • the first and second heavy chain polypeptides form an Fc interface that enhances heterodimerization.
  • (i)-(iv) e.g., nucleic acid encoding (i)-(iv)
  • a single cell e.g., a single mammalian cell, e.g., a CHO cell.
  • (i)-(iv) are expressed in the cell.
  • (i)-(iv) e.g., nucleic acid encoding (i)-(iv)
  • are introduced in different cells e.g., different mammalian cells, e.g., two or more CHO cell.
  • (i)-(iv) are expressed in the cells.
  • the method further comprises purifying a cell-expressed antibody molecule, e.g., using a lambda- and/or- kappa-specific purification, e.g., affinity chromatography .
  • the method further comprises evaluating the cell-expressed multispecific antibody molecule.
  • the purified cell-expressed multispecific antibody molecule can be analyzed by techniques known in the art, include mass spectrometry.
  • the purified cell-expressed antibody molecule is cleaved, e.g., digested with papain to yield the Fab moieties and evaluated using mass spectrometry.
  • the method produces correctly paired kappa/lambda multispecific, e.g., bispecific, antibody molecules in a high yield, e.g., at least 75, 80, 90, 95, 98, 99 99.5 or 99.9%.
  • the multispecific, e.g., a bispecific, antibody molecule that includes:
  • a first heavy chain polypeptide (e.g., a heavy chain polypeptide comprising one, two, three or all of a first heavy chain variable region (first VH), a first CHI, a first heavy
  • T2110-7026WO chain constant region e.g., a first CH2, a first CH3, or both
  • HCP1 binds to a first epitope
  • HCP2 a second heavy chain polypeptide
  • second VH second heavy chain variable region
  • second CHI second heavy chain constant region
  • HCP2 binds to a second epitope
  • LLCP1 lambda light chain polypeptide
  • VL1 lambda light variable region
  • VL1 lambda light constant chain
  • KLCP2 kappa light chain polypeptide
  • VLk lambda light variable region
  • VLk lambda light constant chain
  • the first and second heavy chain polypeptides form an Fc interface that enhances heterodimerization.
  • the multispecific antibody molecule has a first binding specificity that includes a hybrid VL1-CL1 heterodimerized to a first heavy chain variable region connected to the Fc constant, CH2-CH3 domain (having a knob modification) and a second binding specificity that includes a hybrid VLk-CLk heterodimerized to a second heavy chain variable region connected to the Fc constant, CH2-CH3 domain (having a hole modification).
  • CD3-targeting moieties and anti-CD3 antibody molecules CD3-targeting moieties and anti-CD3 antibody molecules
  • the present disclosure provides, inter alia, multispecific (e.g., bi-, tri-, tetra- specific) antibody molecules, that include, e.g., are engineered to contain, one or more CD3-targeting moieties that bind to CD3. Also disclosed herein are anti-CD3 antibody molecules (e.g., monospecific or multispecific antibody molecules that bind to CD3).
  • CD3 refers to cluster of differentiation 3, a protein that, in humans, is composed of four distinct chains (e.g., one CD3y, one CD36, and two CD3s chains) that associate with the T-cell receptor and ⁇ -chain (zeta-chain) to form the TCR complex.
  • CD3 includes, for example, the CD3y, CD3 6, and CD3s chains.
  • UniProt accession numbers P09693, P04234, P07766 provide exemplary human CD3y, CD3 6, and CD3s amino acid sequences, respectively.
  • CD3 or CD3 molecule is a naturally existing CD3 or a functional variant or fragment thereof.
  • the CD3-targeting moiety targets CD3s.
  • the CD3-targeting moiety comprises an antibody, or an antigenbinding fragment thereof.
  • the CD3-targeting moiety comprises a CDR, a framework region, or a variable region sequence described herein (or a sequence having at least about 75%, 80%, 85%, 90%, 95%, or 99% sequence identity thereto).
  • the multispecific antibody molecule comprises a binding domain that targets an immune cell antigen, e.g., an immune effector cell antigen.
  • Immune effector cells include T cells and non-T cell effector cells (e.g., macrophages or natural killer (NK) cells).
  • the multispecific antibody molecule may comprise this binding moiety in addition to, or, in the alternative, in place of the CD3 binding moiety.
  • Non-limiting examples of immune effector cell antigens include the T cell receptor (TCR), CD28, CD16, NKG2D, 0X40, 4-1BB, CD2, CD5, AND CD95.
  • a multispecific antibody molecule comprising a CD3- targeting moiety.
  • an anti-CD3 antibody molecule e.g., a monoclonal anti-CD3 antibody molecule.
  • the CD3-targeting moiety or anti-CD3 antibody molecule comprises an antibody, or an antigen-binding fragment thereof, disclosed in any of Tables 2A- 2D.
  • the CD3-targeting moiety or anti-CD3 antibody molecule comprises a VH comprising a heavy chain complementarity determining region 1 (VHCDR1) of an anti-CD3 antibody molecule disclosed in any of Tables 2A-2D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), a VHCDR2 of an anti- CD3 antibody molecule disclosed in any of Tables 2A-2D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), and/or a VHCDR3 of an anti-CD3 antibody molecule disclosed in any of Tables 2A-2D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions).
  • VHCDR1 heavy chain complementarity determining region 1
  • the CD3-targeting moiety or anti-CD3 antibody molecule comprises a VL comprising a light chain complementarity determining region 1 (VLCDR1) of an anti-CD3 antibody molecule disclosed in any of Tables 2A-2D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), a VLCDR2 of an anti- CD3 antibody molecule disclosed in any of Tables 2A-2D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), and/or a VLCDR3 of an anti-CD3 antibody molecule disclosed in any of Tables 2A-2D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions).
  • VLCDR1 light chain complementarity determining region 1
  • the CD3-targeting moiety or anti-CD3 antibody molecule comprises a VH comprising an amino acid sequence of an anti-CD3 antibody molecule disclosed in any of Tables 2A-2D, or an amino acid sequence having at least about 80%, 85%, 90%, 95%, or 99% sequence identity thereto. In some embodiments, the CD3-targeting moiety or anti-CD3 antibody molecule comprises a VL comprising the amino acid sequence of an anti-CD3 antibody molecule disclosed in any of Tables 2A-2D, or an amino acid sequence having at least about 80%, 85%, 90%, 95%, or 99% sequence identity thereto.
  • the CD3-targeting moiety or anti-CD3 antibody molecule comprises a VH comprising the amino acid sequence of an anti-CD3 antibody molecule disclosed in any of Tables 2A-2D (or an amino acid sequence having at least about 80%, 85%, 90%, 95%, or 99% sequence identity thereto) and a VL comprising the amino acid sequence of an anti-CD3 antibody molecule disclosed in any of Tables 2A-2D (or an amino acid sequence having at least about 80%, 85%, 90%, 95%, or 99% sequence identity thereto).
  • Additional exemplary anti-CD3 antibody molecules are disclosed in WO 1997/044362; WO 2004/106381; WO 2007/009064; WO 2012/162067; WO 2014/141152; WO 2018/224441; US 2014/0302064; US 2003/0108548; US 2007/0224191; US 2018 /0177880; U.S. Pat. Nos. 4,658,019; 6,406,696; 7,381,803; and 10,174,124; and EP Pat. No. 2,155,788 .
  • the multispecific antibody molecule comprises one, two, or three heavy chain CDRs of an anti-CD3 antibody described herein. In an embodiment, the multispecific antibody molecule comprises three heavy chain CDRs of an anti-CD3 antibody described herein. In an embodiment, the multispecific antibody molecule comprises one, two, or three light chain CDRs of an anti-CD3 antibody described herein. In an embodiment, the multispecific antibody molecule comprises three light chain CDRs of an anti-CD3 antibody described herein. In an embodiment, the multispecific antibody molecule comprises one, two, or three heavy chain CDRs and one, two, or three light chain CDRs, of an anti-CD3 antibody described herein. In an embodiment, the multispecific antibody molecule comprises three heavy chain CDRs and three light chain CDRs, of an anti-CD3 antibody described herein.
  • the multispecific antibody molecule comprises a heavy chain variable region of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a heavy chain variable region of an anti-CD3 antibody described herein.
  • the multispecific antibody molecule comprises a light chain variable region of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a light chain variable region of an anti-CD3 antibody described herein.
  • the multispecific antibody molecule comprises a heavy chain variable region of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto, and a light chain variable region of the anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a heavy chain variable region and a light chain variable region, of an anti-CD3 antibody described herein.
  • the multispecific antibody molecule comprises a heavy chain of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a heavy chain of an anti-CD3 antibody described herein.
  • the multispecific antibody molecule comprises a light chain of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a light chain of an anti-CD3 antibody described herein.
  • the multispecific antibody molecule comprises a heavy chain of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto, and a light chain of the anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a heavy chain and a light chain, of an anti-CD3 antibody described herein.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GFSFNIFA (SEQ ID NO: 1); (b) a VHCDR2 comprising the amino acid sequence of IRSKSNNYAT (SEQ ID NO: 2); (c) a VHCDR3 comprising the amino acid sequence of VRQGTGRGGAWFAY (SEQ ID NO: 3); (d) a VLCDR1 comprising the amino acid sequence of QHINNY (SEQ ID NO: 4); (e) a VLCDR2 comprising the amino acid sequence of YTS (SEQ ID NO: 5); and/or (f) a VLCDR3 comprising the amino acid sequence of LQYDNLYT (SEQ ID NO: 6), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GFSFNIFA (SEQ ID NO: 1); (b) a VHCDR2 comprising the amino acid sequence of IRSKSNNYAT (SEQ ID NO: 2); (c) a VHCDR3 comprising the amino acid sequence of VRQGTGRGGAWFAY (SEQ ID NO: 3); (d) a VLCDR1 comprising the amino acid sequence of QHINNY (SEQ ID NO: 4); (e) a VLCDR2 comprising the amino acid sequence
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 7 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 8.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 7 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 8.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 9 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 10.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 1, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GHTFRSSY (SEQ ID NO: 11); (b) a VHCDR2 comprising the amino acid sequence of FYPGSDNA (SEQ ID NO: 12); (c) a VHCDR3 comprising the amino acid sequence of ASDSIMYFFDF (SEQ ID NO: 13); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSFILRT (SEQ ID NO: 16), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GHTFRSSY (SEQ ID NO: 11); (b) a VHCDR2 comprising the amino acid sequence of FYPGSDNA (SEQ ID NO: 12); (c) a VHCDR3 comprising the amino acid sequence of ASDSIMYFFDF (SEQ ID NO: 13); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 17 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 18.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 17 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 18.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 19 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 20.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 2, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTNYW (SEQ ID NO: 21); (b) a VHCDR2 comprising the amino acid sequence of INPNNGGT (SEQ ID NO: 22); (c) a VHCDR3 comprising the amino acid sequence of ARRGNYYHFAY (SEQ ID NO: 23); (d) a VLCDR1 comprising the amino acid sequence of QSIVHSNRYTY (SEQ ID NO: 24); (e) a VLCDR2 comprising the amino acid sequence of GVS (SEQ ID NO: 25); and/or (f) a VLCDR3 comprising the amino acid sequence of FQGTHVPYT (SEQ ID NO: 26), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTNYW (SEQ ID NO: 21); (b) a VHCDR2 comprising the amino acid sequence of INPNNGGT (SEQ ID NO: 22); (c) a VHCDR3 comprising the amino acid sequence of ARRGNYYHFAY (SEQ ID NO: 23); (d) a VLCDR1 comprising the amino acid sequence of QSIVHSNRYTY (SEQ ID NO: 24); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 27 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 28.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 27 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 28.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 29 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 30.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 3, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGGT (SEQ ID NO: 32); (c) a VHCDR3 comprising the amino acid sequence of ARDGFSQYYFDC (SEQ ID NO: 33); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 36), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGGT (SEQ ID NO: 32); (c) a VHCDR3 comprising the amino acid sequence of ARDGFSQYYFDC (SEQ ID NO: 33); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 37 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 38.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 37 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 38.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 39 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 40.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 4, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGVN (SEQ ID NO: 42); (c) a VHCDR3 comprising the amino acid sequence of ARDGSSQYYFDY (SEQ ID NO: 43); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 36), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 41); (b) a VHCDR2 comprising the amino acid sequence of INLSNGVN (SEQ ID NO: 42); (c) a VHCDR3 comprising the amino acid sequence of ARDGSSQYYFDY (SEQ ID NO: 43); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 47 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 48.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 47 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 48.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 49 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 50.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 5, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGVT (SEQ ID NO: 52); (c) a VHCDR3 comprising the amino acid sequence of ARDGSSQYYFDY (SEQ ID NO: 43); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of TQSFILRS (SEQ ID NO: 56), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGVT (SEQ ID NO: 52); (c) a VHCDR3 comprising the amino acid sequence of ARDGSSQYYFDY (SEQ ID NO: 43); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 57 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 58.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 57 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 58.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 59 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 60.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 6, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGGT (SEQ ID NO: 32); (c) a VHCDR3 comprising the amino acid sequence of ARDGTSQYYFDY (SEQ ID NO: 63); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 36), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGGT (SEQ ID NO: 32); (c) a VHCDR3 comprising the amino acid sequence of ARDGTSQYYFDY (SEQ ID NO: 63); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 67 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 68.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 67 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 68. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 69 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 70. In an embodiment, the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 7, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYNFTNFY (SEQ ID NO: 71); (b) a VHCDR2 comprising the amino acid sequence of IFPGSGNT (SEQ ID NO: 72); (c) a VHCDR3 comprising the amino acid sequence of ARNNNYSFDY (SEQ ID NO: 73); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSFILRT (SEQ ID NO: 16), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYNFTNFY (SEQ ID NO: 71); (b) a VHCDR2 comprising the amino acid sequence of IFPGSGNT (SEQ ID NO: 72); (c) a VHCDR3 comprising the amino acid sequence of ARNNNYSFDY (SEQ ID NO: 73); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 77 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 78.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 77 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 78. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 79 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 80. In an embodiment, the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 8, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYIFTSSY (SEQ ID NO: 81); (b) a VHCDR2 comprising the amino acid sequence of IYLGDGNT (SEQ ID NO: 82); (c) a VHCDR3 comprising the amino acid sequence of ARDSYYNYYFDY (SEQ ID NO: 83); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSYNLRT (SEQ ID NO: 86), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10,
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYIFTSSY (SEQ ID NO: 81); (b) a VHCDR2 comprising the amino acid sequence of IYLGDGNT (SEQ ID NO: 82); (c) a VHCDR3 comprising the amino acid sequence of ARDSYYNYYFDY (SEQ ID NO: 83); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 87 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 88.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 87 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 88. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 89 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 90. In an embodiment, the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 9, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 91); (b) a VHCDR2 comprising the amino acid sequence of INLSNGGT (SEQ ID NO: 32); (c) a VHCDR3 comprising the amino acid sequence of ARDGTGQYYFDY (SEQ ID NO: 93); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 36), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 91); (b) a VHCDR2 comprising the amino acid sequence of INLSNGGT (SEQ ID NO:32); (c) a VHCDR3 comprising the amino acid sequence of ARDGTGQYYFDY (SEQ ID NO: 93); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 374 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 38.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 374 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 38.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 375 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 376.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 10, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYIFTSSY (SEQ ID NO: 81); (b) a VHCDR2 comprising the amino acid sequence of IYLGDGNT (SEQ ID NO: 82); (c) a VHCDR3 comprising the amino acid sequence of ARDSYYNYYFDY (SEQ ID NO: 83); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSYNLRT (SEQ ID NO: 86), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10,
  • the multispecific antibody molecule (a) a VHCDR1 comprising the amino acid sequence of GYIFTSSY (SEQ ID NO: 81); (b) a VHCDR2 comprising the amino acid sequence of IYLGDGNT (SEQ ID NO: 82); (c) a VHCDR3 comprising the amino acid sequence of ARDSYYNYYFDY (SEQ ID NO: 83); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of 86
  • WAS (SEQ ID NO: 15); and (f) a VLCDR3 comprising the amino acid sequence of KQSYNLRT (SEQ ID NO: 86), such as possessed by the anti-CD3 antibody Antibody 11.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 87 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 88.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 87 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 88. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 89 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 90. In an embodiment, the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 11, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 13); (b) a VHCDR2 comprising the amino acid sequence of INLSNGDT (SEQ ID NO: 94); (c) a VHCDR3 comprising the amino acid sequence of ARDGISQYYFDF (SEQ ID NO: 95); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of TQSFTLRT (SEQ ID NO: 96), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 13); (b) a VHCDR2 comprising the amino acid sequence of INLSNGDT (SEQ ID NO: 94); (c) a VHCDR3 comprising the amino acid sequence of ARDGISQYYFDF (SEQ ID NO: 95); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid 87
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 118 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 120.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 118 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 120.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 119 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 120.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 12, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GFTFTDYY (SEQ ID NO: 97); (b) a VHCDR2 comprising the amino acid sequence of IRNKANGYTT (SEQ ID NO: 98); (c) a VHCDR3 comprising the amino acid sequence of VYFTLDY (SEQ ID NO: 99); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSYNLYT (SEQ ID NO: 100), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GFTFTDYY (SEQ ID NO: 97); (b) a VHCDR2 comprising the amino acid sequence of IRNKANGYTT (SEQ ID NO: 98); (c) a VHCDR3 comprising the amino acid sequence of VYFTLDY (SEQ ID NO: 99); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of 88
  • WAS (SEQ ID NO: 15); and (f) a VLCDR3 comprising the amino acid sequence of KQSYNLYT (SEQ ID NO: 100), such as possessed by the anti-CD3 antibody Antibody 13.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 122 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 124.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 122 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 124. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 123 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 125. In an embodiment, the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 13, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GHTFRSSY (SEQ ID NO: 11); (b) a VHCDR2 comprising the amino acid sequence of FYPGSDNA (SEQ ID NO: 12); (c) a VHCDR3 comprising the amino acid sequence of ASDSIMYFFDF (SEQ ID NO: 13); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSFILRT (SEQ ID NO: 16), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GHTFRSSY (SEQ ID NO: 11); (b) a VHCDR2 comprising the amino acid sequence of FYPGSDNA (SEQ ID NO: 12); (c) a VHCDR3 comprising the amino acid sequence of ASDSIMYFFDF (SEQ ID NO: 13); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid 89
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 17 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 127.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 17 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 127.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 126 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 128.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 14, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYRFTSYF (SEQ ID NO: 101); (b) a VHCDR2 comprising the amino acid sequence of IYPGSDNI (SEQ ID NO: 102); (c) a VHCDR3 comprising the amino acid sequence of ASDGYVYYFDN (SEQ ID NO: 103); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSFILRT (SEQ ID NO: 16), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYRFTSYF (SEQ ID NO: 101); (b) a VHCDR2 comprising the amino acid sequence of IYPGSDNI (SEQ ID NO: 102); (c) a VHCDR3 comprising the amino acid sequence of ASDGYVYYFDN (SEQ ID NO: 103); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid 90
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 129 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 131.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 129 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 131.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 130 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 132.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 15, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GLNIKDYY (SEQ ID NO: 104); (b) a VHCDR2 comprising the amino acid sequence of IVPGNGNT (SEQ ID NO: 105); (c) a VHCDR3 comprising the amino acid sequence of ARDSYGQYYFDH (SEQ ID NO: 106); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSYTLRT (SEQ ID NO: 107), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10,
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GLNIKDYY (SEQ ID NO: 104); (b) a VHCDR2 comprising the amino acid sequence of IVPGNGNT (SEQ ID NO: 105); (c) a VHCDR3 comprising the amino acid sequence of ARDSYGQYYFDH (SEQ ID NO: 106); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 133 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 135.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 133 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 135.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 134 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 136.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 16, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYSFTSSY (SEQ ID NO: 108); (b) a VHCDR2 comprising the amino acid sequence of IYPGSGNT (SEQ ID NO: 109); (c) a VHCDR3 comprising the amino acid sequence of ARDSIMYYFDY (SEQ ID NO: 110); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSYILRT (SEQ ID NO: 111), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYSFTSSY (SEQ ID NO: 108); (b) a VHCDR2 comprising the amino acid sequence of IYPGSGNT (SEQ ID NO: 109); (c) a VHCDR3 comprising the amino acid sequence of ARDSIMYYFDY (SEQ ID NO: 110); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 137 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 139.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 137 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 139. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 138 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 140. In an embodiment, the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 17, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of IDPNSGDT (SEQ ID NO: 112); (c) a VHCDR3 comprising the amino acid sequence of TRDAYGNYYFDY (SEQ ID NO: 113); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSFILRT (SEQ ID NO: 16), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of IDPNSGDT (SEQ ID NO: 112); (c) a VHCDR3 comprising the amino acid sequence of TRDAYGNYYFDY (SEQ ID NO: 113); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid 93
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 141 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 143.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 141 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 143.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 142 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 144.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 18, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYIFTSSY (SEQ ID NO: 81); (b) a VHCDR2 comprising the amino acid sequence of IYLGDGNT (SEQ ID NO: 82); (c) a VHCDR3 comprising the amino acid sequence of ARDTYYNYYFDY (SEQ ID NO: 114); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSYNLRT (SEQ ID NO: 86), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15,
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYIFTSSY (SEQ ID NO: 81); (b) a VHCDR2 comprising the amino acid sequence of IYLGDGNT (SEQ ID NO: 82); (c) a VHCDR3 comprising the amino acid sequence of ARDTYYNYYFDY (SEQ ID NO: 114); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 145 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 88.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 145 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 88. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 146 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 90. In an embodiment, the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 19, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGGT (SEQ ID NO: 32); (c) a VHCDR3 comprising the amino acid sequence of ARDGSSQYYFDY (SEQ ID NO: 43); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 36), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGGT (SEQ ID NO: 32); (c) a VHCDR3 comprising the amino acid sequence of ARDGSSQYYFDY (SEQ ID NO: 43); (d) a VLCDR1 95
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 147 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 149.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 147 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 149. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 148 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 150. In an embodiment, the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 20, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGVT (SEQ ID NO: 52); (c) a VHCDR3 comprising the amino acid sequence of ARDGTSQYYFDY (SEQ ID NO: 63); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 36), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGVT (SEQ ID NO: 52); (c) a VHCDR3 96
  • T2110-7026WO comprising the amino acid sequence of ARDGTSQYYFDY (SEQ ID NO: 63); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and (f) a VLCDR3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 36), such as possessed by the anti-CD3 antibody Antibody 21.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 151 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 38.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 151 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 38.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 152 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 153.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 21, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of INLSNGGT (SEQ ID NO: 32); (c) a VHCDR3 comprising the amino acid sequence of ARDGSGQYYFDY (SEQ ID NO: 115); (d) a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 36), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids
  • T2110-7026WO comprising the amino acid sequence of INLSNGGT (SEQ ID NO: 32);
  • VHCDR3 comprising the amino acid sequence of ARDGSGQYYFDY (SEQ ID NO: 115);
  • a VLCDR1 comprising the amino acid sequence of QSLLNSRTRKNY (SEQ ID NO: 14);
  • VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15);
  • a VLCDR3 comprising the amino acid sequence of TQSFILRT (SEQ ID NO: 36), such as possessed by the anti-CD3 antibody Antibody 22.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 154 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 156.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 154 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 156. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 155 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 157. In an embodiment, the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 22, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CD3 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYIFTSSY (SEQ ID NO: 81); (b) a VHCDR2 comprising the amino acid sequence of IYLGDGNT (SEQ ID NO: 82); (c) a VHCDR3 comprising the amino acid sequence of ARDSYSNYYFDY (SEQ ID NO: 116); (d) a VLCDR1 comprising the amino acid sequence of QILLNSRTRKNY (SEQ ID NO: 117); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of KQSYTLRT (SEQ ID NO: 107), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 158 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 160.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 158 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 160.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 159 and a VL domain of an anti-CD3 antibody encoded by the nucleic acid sequence of SEQ ID NO: 161.
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 23, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 377 and/or a VL domain of an anti-CD3 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 379.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 377 and a VL domain of an anti-CD3 antibody comprising the amino acid sequence of SEQ ID NO: 379. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD3 antibody encoded by the nucleic acid 99
  • the multispecific antibody molecule comprises the anti-CD3 antibody molecule Antibody 24, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises an anti-CD3 antibody described herein.
  • the multispecific antibody molecule comprises a moiety that binds to the same, or substantially the same, epitope of an anti-CD3 antibody described herein. In an embodiment, the multispecific antibody molecule competes for binding to CD3 with an anti-CD3 antibody described herein.
  • CD117-targeting moieties and anti-CD117 antibody molecules CD117-targeting moieties and anti-CD117 antibody molecules
  • the present disclosure provides, inter alia, multispecific (e.g., bi-, tri-, tetra- specific) antibody molecules, that include, e.g., are engineered to contain, one or more CD117-targeting moieties that bind to CD117. Also disclosed herein are anti-CDl 17 antibody molecules (e.g., monospecific or multispecific antibody molecules that bind to CD117).
  • cKIT refers to mast/stem cell growth factor receptor kit, also known as CD117.
  • Swiss- Prot accession number P 10721 provides exemplary human cKIT amino acid sequences.
  • cKIT or cKIT molecule is a naturally existing cKIT or a functional variant or fragment thereof.
  • the CD 117-targ eting moiety comprises an antibody, or an antigenbinding fragment thereof.
  • the CD 117-targ eting moiety comprises a CDR, a framework region, or a variable region sequence described herein (or a sequence having at least about 75%, 80%, 85%, 90%, 95%, or 99% sequence identity thereto).
  • the multispecific antibody molecule comprises a binding domain that targets a hematopoietic stem cell (HSC) antigen or a cancer cell antigen.
  • HSC hematopoietic stem cell
  • the multispecific antibody molecule may comprise this binding moiety in addition to, or, in the alternative, in place of the CD117 binding moiety.
  • the HSC antigen is CD34.
  • a multispecific antibody molecule comprising a CD117- targeting moiety.
  • an anti-CD117 antibody molecule e.g., a monoclonal anti-CD117 antibody molecule.
  • the CD 117-targ eting moiety or anti-CD117 antibody molecule comprises an antibody, or an antigen-binding fragment thereof, disclosed in any of Tables 3A- 3D.
  • the CD 117-targ eting moiety or anti-CD117 antibody molecule comprises a VH comprising a heavy chain complementarity determining region 1 (VHCDR1) of an anti-CDl 17 antibody molecule disclosed in any of Tables 3A-3D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), a VHCDR2 of an anti- CDl 17 antibody molecule disclosed in any of Tables 3A-3D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), and/or a VHCDR3 of an anti- CDl 17 antibody molecule disclosed in any of Tables 3A-3D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions).
  • VHCDR1 heavy chain complementarity determining region 1
  • the CD 117-targ eting moiety or anti-CDl 17 antibody molecule comprises a VL comprising a light chain complementarity determining region 1 (VLCDR1) of an anti-CDl 17 antibody molecule disclosed in any of Tables 3A-3D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), a VLCDR2 of an anti- CDl 17 antibody molecule disclosed in any of Tables 3A-3D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), and/or a VLCDR3 of an anti- CDl 17 antibody molecule disclosed in any of Tables 3A-3D (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions).
  • VLCDR1 light chain complementarity determining region 1
  • the CD 117-targ eting moiety or anti-CDl 17 antibody molecule comprises a VH comprising an amino acid sequence of an anti-CDl 17 antibody molecule disclosed in any of Tables 3A-3D, or an amino acid sequence having at least about 80%, 85%, 90%, 95%, or 99% sequence identity thereto.
  • the CD117-targeting moiety or anti-CDl 17 antibody molecule comprises a VL comprising the amino acid sequence of an anti-CDl 17 antibody molecule disclosed in any of Tables 3A-3D, or an amino acid sequence having at least about 80%, 85%, 90%, 95%, or 99% sequence identity thereto.
  • the CD 117-targ eting moiety or anti-CD117 antibody molecule comprises a VH comprising the amino acid sequence of an anti-CD117 antibody molecule disclosed in any of Tables 3A-3D (or an amino acid sequence having at least about 80%, 85%, 90%, 95%, or 99% sequence identity thereto) and a VL comprising the amino acid sequence of an anti-CDl 17 antibody molecule disclosed in any of Tables 3A-3D (or an amino acid sequence having at least about 80%, 85%, 90%, 95%, or 99% sequence identity thereto).
  • Table 3A Exemplary CD117 Antibody Molecules Table 3B: Exemplary CD117 Antibody Sequences
  • Additional exemplary anti-CD117 antibody molecules are disclosed in WO 2007/127317; WO 2012/103165; WO 2014/150937; WO 2014/018625; WO 2015/067667; WO 2016/020791; WO 2017/219029; WO 2018/116178; WO 2019/084057; WO 2019/084064; and WO 2019/084067.
  • the multispecific antibody molecule comprises one, two, or three heavy chain CDRs of an anti-CDl 17 antibody described herein. In an embodiment, the multispecific antibody molecule comprises three heavy chain CDRs of an anti-CDl 17 antibody described herein. In an embodiment, the multispecific antibody molecule comprises one, two, or three light chain CDRs of an anti-CDl 17 antibody described herein. In an embodiment, the multispecific antibody molecule comprises three light chain CDRs of an anti-CDl 17 antibody described herein. In an embodiment, the multispecific antibody molecule comprises one, two, or three heavy chain CDRs and one, two, or three light chain CDRs, of an anti-CDl 17 antibody described herein. In an embodiment, the multispecific antibody molecule comprises three heavy chain CDRs and three light chain CDRs, of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises a heavy chain variable region of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a heavy chain variable region of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises a light chain variable region of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a light chain variable region of an anti-CDl 17 antibody described herein. In an embodiment, the multispecific antibody molecule comprises a heavy chain variable region of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 134
  • the multispecific antibody molecule comprises a heavy chain variable region and a light chain variable region, of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises a heavy chain of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a heavy chain of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises a light chain of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a light chain of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises a heavy chain of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto, and a light chain of the anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises a heavy chain and a light chain, of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises a moiety that binds to the same, or substantially the same, epitope of an anti-CDl 17 antibody described herein. In an embodiment, the multispecific antibody molecule competes for binding to CD3 with an antiCD 117 antibody described herein.
  • the multispecific antibody molecule comprises (a) one, two, or three heavy chain CDRs of an anti-CD3 antibody described herein and (b) one, two, or three heavy chain CDRs of an anti-CDl 17 antibody described herein. In an embodiment, the multispecific antibody molecule comprises (a) three heavy chain CDRs of an anti-CD3 antibody described herein and (b) three heavy chain CDRs of an anti-CDl 17 antibody described herein. In an 135
  • the multispecific antibody molecule comprises (a) one, two, or three light chain CDRs of an anti-CD3 antibody described herein and (b) one, two, or three light chain CDRs of an anti-CD117 antibody described herein.
  • the multispecific antibody molecule comprises (a) three light chain CDRs of an anti-CD3 antibody described herein and (b) three light chain CDRs of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises (a) one, two, or three heavy chain CDRs and one, two, or three light chain CDRs, of an anti-CD3 antibody described herein and (b) one, two, or three heavy chain CDRs and one, two, or three light chain CDRs, of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises (a) three heavy chain CDRs and three light chain CDRs, of an anti-CD3 antibody described herein and (b) three heavy chain CDRs and three light chain CDRs, of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises (a) a heavy chain variable region of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto and (b) a heavy chain variable region of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises (a) a heavy chain variable region of an anti-CD3 antibody described herein and (b) a heavy chain variable region of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises (a) a light chain variable region of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto and (b) a light chain variable region of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises (a) a light chain variable region of an anti-CD3 antibody described herein and (b) a light chain variable region of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises (a) a heavy chain variable region of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto, and a
  • the multispecific antibody molecule comprises (a) a heavy chain variable region and a light chain variable region, of an anti-CD3 antibody described herein and (b) a heavy chain variable region and a light chain variable region, of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises (a) a heavy chain of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto and (b) a heavy chain of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises (a) a heavy chain of an anti-CD3 antibody described herein and (b) a heavy chain of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises (a) a light chain of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto and (b) a light chain of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto.
  • the multispecific antibody molecule comprises (a) a light chain of an anti-CD3 antibody described herein and (b) a light chain of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises (a) a heavy chain of an anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto, and a light chain of the anti-CD3 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 90%, 95%, 98%, or 99% identical thereto and (b) a heavy chain of an anti-CDl 17 antibody described herein, or a sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids therefrom, or at least 85%, 137
  • the multispecific antibody molecule comprises (a) a heavy chain and a light chain, of an anti-CD3 antibody described herein and (b) a heavy chain and a light chain, of an anti-CDl 17 antibody described herein.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTNYW (SEQ ID NO: 21); (b) a VHCDR2 comprising the amino acid sequence of INPSNGRT (SEQ ID NO: 162); (c) a VHCDR3 comprising the amino acid sequence of ARSAIVTNWYFDV (SEQ ID NO: 163); (d) a VLCDR1 comprising the amino acid sequence of GNVHNY (SEQ ID NO: 166); (e) a VLCDR2 comprising the amino acid sequence of YVK (SEQ ID NO: 167); and/or (f) a VLCDR3 comprising the amino acid sequence of QHFWNTPYT (SEQ ID NO: 168), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTNYW (SEQ ID NO: 21); (b) a VHCDR2 comprising the amino acid sequence of INPSNGRT (SEQ ID NO: 162); (c) a VHCDR3 comprising the amino acid sequence of ARSAIVTNWYFDV (SEQ ID NO: 163); (d) a VLCDR1 comprising the amino acid sequence of GNVHNY (SEQ ID NO: 166); (e) a VLCDR2 comprising the amino acid sequence of YVK (SEQ ID NO: 167); and/or (f) a VLCDR3 comprising the amino acid sequence of QHFWNTPYT (SEQ ID NO: 168), such as possessed by the anti-CDl 17 antibody Antibody 25.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 164 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 169.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 138
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody encoded by the nucleic acid sequence of SEQ ID NO: 165 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 170.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 25, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GFTFSGYG (SEQ ID NO: 171); (b) a VHCDR2 comprising the amino acid sequence of ISSSSGNI (SEQ ID NO: 172); (c) a VHCDR3 comprising the amino acid sequence of VRSYSLDY (SEQ ID NO: 173); (d) a VLCDR1 comprising the amino acid sequence of SSVSSTY (SEQ ID NO: 176); (e) a VLCDR2 comprising the amino acid sequence of STS (SEQ ID NO: 177); and/or (f) a VLCDR3 comprising the amino acid sequence of HQYHRSPPT (SEQ ID NO: 178), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GFTFSGYG (SEQ ID NO: 171); (b) a VHCDR2 comprising the amino acid sequence of ISSSSGNI (SEQ ID NO: 172); (c) a VHCDR3 comprising the amino acid sequence of VRSYSLDY (SEQ ID NO: 173); (d) a VLCDR1 comprising the amino acid sequence of SSVSSTY (SEQ ID NO: 176); (e) a VLCDR2 comprising the amino acid sequence of STS (SEQ ID NO: 177); and/or (f) a VLCDR3 comprising the amino acid sequence of HQYHRSPPT (SEQ ID NO: 178), such as possessed by the anti-CDl 17 antibody Antibody 26.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 174 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 179.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 139
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody encoded by the nucleic acid sequence of SEQ ID NO: 175 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 180.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 26, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYIFTSYG (SEQ ID NO: 181); (b) a VHCDR2 comprising the amino acid sequence of IYPRSGKT (SEQ ID NO: 182); (c) a VHCDR3 comprising the amino acid sequence of ARNSGYDEGFAY (SEQ ID NO: 183); (d) a VLCDR1 comprising the amino acid sequence of VNIYSY (SEQ ID NO: 186); (e) a VLCDR2 comprising the amino acid sequence of NTK (SEQ ID NO: 187); and/or (f) a VLCDR3 comprising the amino acid sequence of QSHYDTPRT (SEQ ID NO: 188), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYIFTSYG (SEQ ID NO: 181); (b) a VHCDR2 comprising the amino acid sequence of IYPRSGKT (SEQ ID NO: 182); (c) a VHCDR3 comprising the amino acid sequence of ARNSGYDEGFAY (SEQ ID NO: 183); (d) a VLCDR1 comprising the amino acid sequence of VNIYSY (SEQ ID NO: 186); (e) a VLCDR2 comprising the amino acid sequence of NTK (SEQ ID NO: 187); and/or (f) a VLCDR3 comprising the amino acid sequence of QSHYDTPRT (SEQ ID NO: 188), such as possessed by the anti-CDl 17 antibody Antibody 27.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 184 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 189.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 140
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody encoded by the nucleic acid sequence of SEQ ID NO: 185 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 190.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 27, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTNYW (SEQ ID NO: 21); (b) a VHCDR2 comprising the amino acid sequence of IDPSDSYT (SEQ ID NO: 191); (c) a VHCDR3 comprising the amino acid sequence of ARSRYTMDY (SEQ ID NO: 192); (d) a VLCDR1 comprising the amino acid sequence of SSVSY (SEQ ID NO: 195); (e) a VLCDR2 comprising the amino acid sequence of STS (SEQ ID NO: 177); and/or (f) a VLCDR3 comprising the amino acid sequence of HQGSSYPLT (SEQ ID NO: 196), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTNYW (SEQ ID NO: 21); (b) a VHCDR2 comprising the amino acid sequence of IDPSDSYT (SEQ ID NO: 191); (c) a VHCDR3 comprising the amino acid sequence of ARSRYTMDY (SEQ ID NO: 192); (d) a VLCDR1 comprising the amino acid sequence of SSVSY (SEQ ID NO: 195); (e) a VLCDR2 comprising the amino acid sequence of STS (SEQ ID NO: 177); and/or (f) a VLCDR3 comprising the amino acid sequence of HQGSSYPLT (SEQ ID NO: 196), such as possessed by the anti-CDl 17 antibody Antibody 28.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 193 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 197.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising the amino acid sequence of SEQ ID NO: 193 and a VL domain of an anti- CD117 antibody comprising the amino acid sequence of SEQ ID NO: 197.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody encoded by the nucleic acid sequence of SEQ ID NO: 194 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 198.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 28, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GFSLTSYN (SEQ ID NO: 199); (b) a VHCDR2 comprising the amino acid sequence of IWAGGNT (SEQ ID NO: 200); (c) a VHCDR3 comprising the amino acid sequence of ANSKGAWFAY (SEQ ID NO: 201); (d) a VLCDR1 comprising the amino acid sequence of QDVGTA (SEQ ID NO: 204); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of QQYGSYPLP (SEQ ID NO: 205), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GFSLTSYN (SEQ ID NO: 199); (b) a VHCDR2 comprising the amino acid sequence of IWAGGNT (SEQ ID NO: 200); (c) a VHCDR3 comprising the amino acid sequence of ANSKGAWFAY (SEQ ID NO: 201); (d) a VLCDR1 comprising the amino acid sequence of QDVGTA (SEQ ID NO: 204); (e) a VLCDR2 comprising the amino acid sequence of WAS (SEQ ID NO: 15); and/or (f) a VLCDR3 comprising the amino acid sequence of QQYGSYPLP (SEQ ID NO: 205), such as possessed by the anti-CDl 17 antibody Antibody 29.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 202 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 142 4374195.1 Attorney Docket No.: T2110-7026WO
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising the amino acid sequence of SEQ ID NO: 202 and a VL domain of an antiCD 117 antibody comprising the amino acid sequence of SEQ ID NO: 206.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody encoded by the nucleic acid sequence of SEQ ID NO: 203 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 207.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 29, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTEYT (SEQ ID NO: 208); (b) a VHCDR2 comprising the amino acid sequence of FYPGRSTI (SEQ ID NO: 209); (c) a VHCDR3 comprising the amino acid sequence of ARHEEAGRGAFAY (SEQ ID NO: 210); (d) a VLCDR1 comprising the amino acid sequence of QNINVW (SEQ ID NO: 213); (e) a VLCDR2 comprising the amino acid sequence of KAS (SEQ ID NO: 214); and/or (f) a VLCDR3 comprising the amino acid sequence of QQGQSFPYT (SEQ ID NO: 215), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTEYT (SEQ ID NO: 208); (b) a VHCDR2 comprising the amino acid sequence of FYPGRSTI (SEQ ID NO: 209); (c) a VHCDR3 comprising the amino acid sequence of ARHEEAGRGAFAY (SEQ ID NO: 210); (d) a VLCDR1 comprising the amino acid sequence of QNINVW (SEQ ID NO: 213); (e) a VLCDR2 comprising the amino acid sequence of KAS (SEQ ID NO: 214); and/or (f) a VLCDR3 comprising the amino acid sequence of QQGQSFPYT (SEQ ID NO: 215), such as possessed by the anti-CDl 17 antibody Antibody 30.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 211 and/or a VL domain of an anti-CDl 17 antibody comprising an 143
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising the amino acid sequence of SEQ ID NO: 211 and a VL domain of an anti- CD117 antibody comprising the amino acid sequence of SEQ ID NO: 216.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody encoded by the nucleic acid sequence of SEQ ID NO: 212 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 217.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 30, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYSITSGYS (SEQ ID NO: 218); (b) a VHCDR2 comprising the amino acid sequence of IHYSGNT (SEQ ID NO: 219); (c) a VHCDR3 comprising the amino acid sequence of ARPHYISISYWYFDV (SEQ ID NO: 220); (d) a VLCDR1 comprising the amino acid sequence of KSVSTSGYSY (SEQ ID NO: 223); (e) a VLCDR2 comprising the amino acid sequence of LAS (SEQ ID NO: 224 and/or (f) a VLCDR3 comprising the amino acid sequence of QHSRELPPT (SEQ ID NO: 225), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5,
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYSITSGYS (SEQ ID NO: 218); (b) a VHCDR2 comprising the amino acid sequence of IHYSGNT (SEQ ID NO: 219); (c) a VHCDR3 comprising the amino acid sequence of ARPHYISISYWYFDV (SEQ ID NO: 220); (d) a VLCDR1 comprising the amino acid sequence of KSVSTSGYSY (SEQ ID NO: 223); (e) a VLCDR2 comprising the amino acid sequence of LAS (SEQ ID NO: 224 and/or (f) a VLCDR3 comprising the amino acid sequence of QHSRELPPT (SEQ ID NO: 225), such as possessed by the anti-CDl 17 antibody Antibody 31.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 221 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 226.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 222 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 227.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 31, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYW (SEQ ID NO: 228); (b) a VHCDR2 comprising the amino acid sequence of IDTSDSYT (SEQ ID NO: 229); (c) a VHCDR3 comprising the amino acid sequence of ARSGIYSNYGYFDV (SEQ ID NO: 230); (d) a VLCDR1 comprising the amino acid sequence of HDVSTD (SEQ ID NO: 233); (e) a VLCDR2 comprising the amino acid sequence of SAS (SEQ ID NO: 234); and/or (f) a VLCDR3 comprising the amino acid sequence of QQHYIRPRT (SEQ ID NO: 235), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYW (SEQ ID NO: 228); (b) a VHCDR2 comprising the amino acid sequence of IDTSDSYT (SEQ ID NO: 229); (c) a VHCDR3 comprising the amino acid sequence of ARSGIYSNYGYFDV (SEQ ID NO: 230); (d) a VLCDR1 comprising the amino acid sequence of HDVSTD (SEQ ID NO: 233); (e) a VLCDR2 comprising the amino acid sequence of SAS (SEQ ID NO: 234); and/or (f) a VLCDR3 comprising the amino acid sequence of QQHYIRPRT (SEQ ID NO: 235), such as possessed by the anti-CDl 17 antibody Antibody 32.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 145 4374195.1 Attorney Docket No.: T2110-7026WO
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising the amino acid sequence of SEQ ID NO: 231 and a VL domain of an antiCD 117 antibody comprising the amino acid sequence of SEQ ID NO: 236.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody encoded by the nucleic acid sequence of SEQ ID NO: 232 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 237.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 32, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYG (SEQ ID NO: 238); (b) a VHCDR2 comprising the amino acid sequence of INTSTGKP (SEQ ID NO: 239); (c) a VHCDR3 comprising the amino acid sequence of VRRTTVVDAY (SEQ ID NO: 240); (d) a VLCDR1 comprising the amino acid sequence of SSVTY (SEQ ID NO: 243); (e) a VLCDR2 comprising the amino acid sequence of STS (SEQ ID NO: 177); and/or (f) a VLCDR3 comprising the amino acid sequence of QQRNNYPPMYT (SEQ ID NO: 244), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYG (SEQ ID NO: 238); (b) a VHCDR2 comprising the amino acid sequence of INTSTGKP (SEQ ID NO: 239); (c) a VHCDR3 comprising the amino acid sequence of VRRTTVVDAY (SEQ ID NO: 240); (d) a VLCDR1 comprising the amino acid sequence of SSVTY (SEQ ID NO: 243); (e) a VLCDR2 comprising the amino acid sequence of STS (SEQ ID NO: 177); and/or (f) a VLCDR3 comprising the amino acid sequence of QQRNNYPPMYT (SEQ ID NO: 244), such as possessed by the anti-CDl 17 antibody Antibody 33.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 241 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 245.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 241 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 245.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 242 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 246.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 33, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTNYW (SEQ ID NO: 21); (b) a VHCDR2 comprising the amino acid sequence of IDPSDSYT (SEQ ID NO: 191); (c) a VHCDR3 comprising the amino acid sequence of TRSAFGFAY (SEQ ID NO: 247); (d) a VLCDR1 comprising the amino acid sequence of QDVTTA (SEQ ID NO: 250); (e) a VLCDR2 comprising the amino acid sequence of SAS (SEQ ID NO: 234); and/or (f) a VLCDR3 comprising the amino acid sequence of QQHYIIPWT (SEQ ID NO: 251), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from,
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 248 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 252.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 248 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 252.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 249 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 253.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 34, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTTYW (SEQ ID NO: 254); (b) a VHCDR2 comprising the amino acid sequence of IDPNGGDT (SEQ ID NO: 255); (c) a VHCDR3 comprising the amino acid sequence of VRNGWDYAMDS (SEQ ID NO: 256); (d) a VLCDR1 comprising the amino acid sequence of KSLLHSNGNTY (SEQ ID NO: 259); (e) a VLCDR2 comprising the amino acid sequence of RMS (SEQ ID NO: 260); and/or (f) a VLCDR3 comprising the amino acid sequence of MQNLEYPFT (SEQ ID NO: 261), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15,
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTTYW (SEQ ID NO: 254); (b) a VHCDR2 comprising the amino acid sequence of IDPNGGDT (SEQ ID NO: 255); (c) a VHCDR3 comprising the amino acid sequence of VRNGWDYAMDS (SEQ ID NO: 256); (d) a VLCDR1 comprising the amino acid sequence of KSLLHSNGNTY (SEQ ID NO: 259); (e) a VLCDR2 comprising the amino acid 148
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 257 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 262.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 257 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 262.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 258 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 263.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 35, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYSFTVYY (SEQ ID NO: 264); (b) a VHCDR2 comprising the amino acid sequence of IIPYTGGS (SEQ ID NO: 265); (c) a VHCDR3 comprising the amino acid sequence of ARGLGRRYFDV (SEQ ID NO: 266); (d) a VLCDR1 comprising the amino acid sequence of SSVNY (SEQ ID NO: 269); (e) a VLCDR2 comprising the amino acid sequence of DTS (SEQ ID NO: 270); and/or (f) a VLCDR3 comprising the amino acid sequence of QQWRSYPPT (SEQ ID NO: 271), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYSFTVYY (SEQ ID NO: 264); (b) a VHCDR2 comprising the amino acid sequence of IIPYTGGS (SEQ ID NO: 265); (c) a VHCDR3 comprising the amino acid sequence of ARGLGRRYFDV (SEQ ID NO: 266); (d) a VLCDR1 comprising the amino acid 149
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 267 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 272.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 267 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 272.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 268 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 273.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 36, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTNYW (SEQ ID NO: 21); (b) a VHCDR2 comprising the amino acid sequence of IDPNSGDT (SEQ ID NO: 112); (c) a VHCDR3 comprising the amino acid sequence of AKGNFYGSSPLDY (SEQ ID NO: 274); (d) a VLCDR1 comprising the amino acid sequence of EDIYNR (SEQ ID NO: 277); (e) a VLCDR2 comprising the amino acid sequence of GAT (SEQ ID NO: 278); and/or (f) a VLCDR3 comprising the amino acid sequence of QQYWNTPPT (SEQ ID NO: 279), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20
  • the multispecific antibody molecule comprises (a) ) a VHCDR1 comprising the amino acid sequence of GYTFTNYW (SEQ ID NO: 21); (b) a VHCDR2 comprising the amino acid sequence of IDPNSGDT (SEQ ID NO: 112); (c) a VHCDR3 comprising the amino acid sequence of AKGNFYGSSPLDY (SEQ ID NO: 274); (d) a VLCDR1 150
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 275 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 280.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 275 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 280.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 276 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 281.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 37, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTNSW (SEQ ID NO: 282); (b) a VHCDR2 comprising the amino acid sequence of INPSSGFT (SEQ ID NO: 283); (c) a VHCDR3 comprising the amino acid sequence of ARGGVVVTGRWYFDV (SEQ ID NO: 284); (d) a VLCDR1 comprising the amino acid sequence of QNVGPA (SEQ ID NO: 287); (e) a VLCDR2 comprising the amino acid sequence of SAS (SEQ ID NO: 234); and/or (f) a VLCDR3 comprising the amino acid sequence of QQYSSYPLT (SEQ ID NO: 288), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTNSW (SEQ ID NO: 282); (b) a VHCDR2 comprising the amino acid sequence of INPSSGFT (SEQ ID NO: 283); (c) a VHCDR3 151
  • T2110-7026WO comprising the amino acid sequence of ARGGVVVTGRWYFDV (SEQ ID NO: 284); (d) a VLCDR1 comprising the amino acid sequence of QNVGPA (SEQ ID NO: 287); (e) a VLCDR2 comprising the amino acid sequence of SAS (SEQ ID NO: 234); and/or (f) a VLCDR3 comprising the amino acid sequence of QQYSSYPLT (SEQ ID NO: 288), such as possessed by the anti-CD117 antibody Antibody 38.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 285 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 289.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 285 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 289.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 286 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 290.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 38, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GFSLTNYG (SEQ ID NO: 291); (b) a VHCDR2 comprising the amino acid sequence of IWAGGIT (SEQ ID NO: 292); (c) a VHCDR3 comprising the amino acid sequence of VRERGVFDY (SEQ ID NO: 293); (d) a VLCDR1 comprising the amino acid sequence of QNVRTS (SEQ ID NO: 296); (e) a VLCDR2 comprising the amino acid sequence of LAS (SEQ ID NO: 297); and/or (f) a VLCDR3 comprising the amino acid sequence of LQHWNYPLT (SEQ ID NO: 298), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 294 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 299.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 294 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 299.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 295 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 300.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 39, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYY (SEQ ID NO: 301); (b) a VHCDR2 comprising the amino acid sequence of IYPGSGNT (SEQ ID NO: 109); (c) a VHCDR3 comprising the amino acid sequence of ARGVYYFDF (SEQ ID NO: 302); (d) a VLCDR1 comprising the amino acid sequence of QDVRTA (SEQ ID NO: 305); (e) a VLCDR2 comprising the amino acid sequence of SAS (SEQ ID NO: 234); and/or (f) a VLCDR3 comprising the amino acid sequence of QQHSRIPRT (SEQ ID NO: 306), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from
  • T2110-7026WO comprising the amino acid sequence of IYPGSGNT (SEQ ID NO: 109); (c) a VHCDR3 comprising the amino acid sequence of ARGVYYFDF (SEQ ID NO: 302); (d) a VECDR1 comprising the amino acid sequence of QDVRTA (SEQ ID NO: 305); (e) a VLCDR2 comprising the amino acid sequence of SAS (SEQ ID NO: 234); and/or (f) a VLCDR3 comprising the amino acid sequence of QQHSRIPRT (SEQ ID NO: 306), such as possessed by the anti-CD117 antibody Antibody 40.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 303 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 307.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 303 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 307.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 304 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 308.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 40, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYSITSGYY (SEQ ID NO: 309); (b) a VHCDR2 comprising the amino acid sequence of ISYEGSN (SEQ ID NO: 310); (c) a VHCDR3 comprising the amino acid sequence of AREGDYPWFAY (SEQ ID NO: 311); (d) a VLCDR1 comprising the amino acid sequence of QDISNY (SEQ ID NO: 314); (e) a VLCDR2 comprising the amino acid sequence of YTS (SEQ ID NO: 5); and/or (f) a VLCDR3 comprising the amino acid sequence of QQGNTLPYT (SEQ ID NO: 315), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 312 and/or a VL domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 316.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising the amino acid sequence of SEQ ID NO: 312 and a VL domain of an anti- CD117 antibody comprising the amino acid sequence of SEQ ID NO: 316. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody encoded by the nucleic acid sequence of SEQ ID NO: 313 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 317. In an embodiment, the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 41, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTRYY (SEQ ID NO: 318); (b) a VHCDR2 comprising the amino acid sequence of IYPGNDNT (SEQ ID NO: 319); (c) a VHCDR3 comprising the amino acid sequence of ARRGDSYAMDY (SEQ ID NO: 320); (d) a VLCDR1 comprising the amino acid sequence of SSVSSSF (SEQ ID NO: 323); (e) a VLCDR2 comprising the amino acid sequence of STS (SEQ ID NO: 177); and/or (f) a VLCDR3 comprising the amino acid sequence of HQYHRSPYT (SEQ ID NO: 324), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTRYY (SEQ ID NO: 318); (b) a VHCDR2 comprising the amino acid sequence of IYPGNDNT (SEQ ID NO: 319); (c) a VHCDR3 comprising the amino acid sequence of ARRGDSYAMDY (SEQ ID NO: 320); (d) a VLCDR1 comprising the amino acid sequence of SSVSSSF (SEQ ID NO: 323); (e) a VLCDR2 comprising the amino acid sequence of STS (SEQ ID NO: 177); and/or (f) a VLCDR3 comprising the amino acid sequence of HQYHRSPYT (SEQ ID NO: 324), such as possessed by the anti-CD117 antibody Antibody 42.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 321 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 325.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 321 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 325. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 322 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 326. In an embodiment, the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 42, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYS (SEQ ID NO: 327); (b) a VHCDR2 comprising the amino acid sequence of INTETGEP (SEQ ID NO: 328); (c) a VHCDR3 comprising the amino acid sequence of GRGDYPYSFDY (SEQ ID NO: 329); (d) a VLCDR1 comprising the amino acid sequence of GNIHNY (SEQ ID NO: 332); (e) a VLCDR2 comprising the amino acid sequence of NAK (SEQ ID NO: 333); and/or (f) a VLCDR3 comprising the amino acid sequence of HHFWSTPWT (SEQ ID NO: 334), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino
  • the multispecific antibody molecule comprises (a) (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYS (SEQ ID NO: 327); (b) a VHCDR2 comprising the amino acid sequence of INTETGEP (SEQ ID NO: 328); (c) a VHCDR3 comprising the amino acid sequence of GRGDYPYSFDY (SEQ ID NO: 329); (d) a VLCDR1 comprising the amino acid sequence of GNIHNY (SEQ ID NO: 332); (e) a VLCDR2 comprising the amino acid sequence of NAK (SEQ ID NO: 333); and/or (f) a VLCDR3 comprising the amino acid sequence of HHFWSTPWT (SEQ ID NO: 334), such as possessed by the
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 330 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 335.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 330 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 335. In an embodiment, the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 331 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 336. In an embodiment, the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 43, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFLSYW (SEQ ID NO: 337); (b) a VHCDR2 comprising the amino acid sequence of IFPASGST (SEQ ID NO: 338); (c) a VHCDR3 comprising the amino acid sequence of ARSYGNLYYFDY (SEQ ID NO: 339); (d) a VLCDR1 comprising the amino acid sequence of KSLLHSDGITY (SEQ ID NO: 342); (e) a VLCDR2 comprising the amino acid sequence of RMS (SEQ ID NO: 260); and/or (f) a VLCDR3 comprising the amino acid sequence of AQMLEFPFT (SEQ ID NO: 343), or a combination of one or more of the above CDRs and 157
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFLSYW (SEQ ID NO: 337); (b) a VHCDR2 comprising the amino acid sequence of IFPASGST (SEQ ID NO: 338); (c) a VHCDR3 comprising the amino acid sequence of ARSYGNLYYFDY (SEQ ID NO: 339); (d) a VLCDR1 comprising the amino acid sequence of KSLLHSDGITY (SEQ ID NO: 342); (e) a VLCDR2 comprising the amino acid sequence of RMS (SEQ ID NO: 260); and/or (f) a VLCDR3 comprising the amino acid sequence of AQMLEFPFT (SEQ ID NO: 343), such as possessed by the anti-CD117 antibody Antibody 44.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 340 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 344.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 340 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 344.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 341 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 345.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 44, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYN (SEQ ID NO: 346); (b) a VHCDR2 comprising the amino acid sequence of IYPNNGDT (SEQ ID NO: 347); (c) a VHCDR3 comprising the amino acid sequence of AKANGGDGGWYFDF (SEQ ID NO: 348); (d) a VLCDR1 comprising the amino acid sequence of QDISNY (SEQ ID NO: 314); (e) a VLCDR2 comprising the amino acid sequence of YTS (SEQ ID NO: 5); and/or (f) a VLCDR3 comprising the amino acid sequence of 158
  • QQGNTLPWT (SEQ ID NO: 351), or a combination of one or more of the above CDRs and one or more variants thereof differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to any one of SEQ ID NOs: 346-348, 314, 5, or 351.
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYN (SEQ ID NO: 346); (b) a VHCDR2 comprising the amino acid sequence of IYPNNGDT (SEQ ID NO: 347); (c) a VHCDR3 comprising the amino acid sequence of AKANGGDGGWYFDF (SEQ ID NO: 348); (d) a VLCDR1 comprising the amino acid sequence of QDISNY (SEQ ID NO: 314); (e) a VLCDR2 comprising the amino acid sequence of YTS (SEQ ID NO: 5); and/or (f) a VLCDR3 comprising the amino acid sequence of QQGNTLPWT (SEQ ID NO: 351), such as possessed by the antiCD 117 antibody Antibody 45.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 349 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 352.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 349 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 352.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 350 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 353.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 45, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of VDPDSGDT (SEQ ID NO: 354); (c) a VHCDR3 comprising the amino acid sequence of TRDYYDDNYRGHRYYYAMYY (SEQ ID NO: 355); (d) a VLCDR1 comprising the amino acid sequence of KSLLHSNGNTY (SEQ ID NO: 259); (e) a VLCDR2 comprising the 159
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTSYW (SEQ ID NO: 31); (b) a VHCDR2 comprising the amino acid sequence of VDPDSGDT (SEQ ID NO: 354); (c) a VHCDR3 comprising the amino acid sequence of TRDYYDDNYRGHRYYYAMYY (SEQ ID NO: 355); (d) a VLCDR1 comprising the amino acid sequence of KSLLHSNGNTY (SEQ ID NO: 259); (e) a VLCDR2 comprising the amino acid sequence of RMS (SEQ ID NO: 260); and/or (f) a VLCDR3 comprising the amino acid sequence of MQHLEYPFT (SEQ ID NO: 358), such as possessed by the anti-CD117 antibody Antibody 46.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 356 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 359.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 356 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 359.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 357 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 360.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 46, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GFNIKNTY (SEQ ID NO: 361); (b) a VHCDR2 comprising the amino acid sequence of IDPANGNS (SEQ ID NO: 362); (c) a VHCDR3 comprising the amino acid sequence of ASPPYYDGSSYDYAMDN (SEQ ID NO: 363); (d) a VLCDR1 comprising the 160
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GFNIKNTY (SEQ ID NO: 361); (b) a VHCDR2 comprising the amino acid sequence of IDPANGNS (SEQ ID NO: 362); (c) a VHCDR3 comprising the amino acid sequence of ASPPYYDGSSYDYAMDN (SEQ ID NO: 363); (d) a VLCDR1 comprising the amino acid sequence of QDIRIY (SEQ ID NO: 366); (e) a VLCDR2 comprising the amino acid sequence of YTS (SEQ ID NO: 5); and/or (f) a VLCDR3 comprising the amino acid sequence of QQYSKFPWT (SEQ ID NO: 367), such as possessed by the antiCD 117 antibody Antibody 47.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 364 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 368.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 364 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 368.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 365 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 369.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 47, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises at least one, two, three, four, five, or six CDRs of an anti-CDl 17 antibody selected from (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYG (SEQ ID NO: 238); (b) a VHCDR2 comprising the amino acid sequence of INTSTGKP (SEQ ID NO: 239); (c) a VHCDR3 comprising the amino acid 161
  • the multispecific antibody molecule comprises (a) a VHCDR1 comprising the amino acid sequence of GYTFTDYG (SEQ ID NO: 238); (b) a VHCDR2 comprising the amino acid sequence of INTSTGKP (SEQ ID NO: 239); (c) a VHCDR3 comprising the amino acid sequence of VRRTTVVDAY (SEQ ID NO: 240); (d) a VLCDR1 comprising the amino acid sequence of SSVSY (SEQ ID NO: 195); (e) a VLCDR2 comprising the amino acid sequence of STS (SEQ ID NO: 177); and/or (f) a VLCDR3 comprising the amino acid sequence of QQRNNYPPMYT (SEQ ID NO: 244), such as possessed by the anti-CD117 antibody Antibody 48.
  • the multispecific antibody molecule comprises a VH domain of an anti-CD117 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 370 and/or a VL domain of an anti-CDl 17 antibody comprising an amino acid sequence differing by no more than 1, 2, 5, 10, 15, or 20 amino acids from, or at least 85%, 90%, 95%, 98%, or 99% identical to, or the sequence of, SEQ ID NO: 372.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 370 and a VL domain of an anti- CDl 17 antibody comprising the amino acid sequence of SEQ ID NO: 372.
  • the multispecific antibody molecule comprises a VH domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 371 and a VL domain of an anti-CDl 17 antibody encoded by the nucleic acid sequence of SEQ ID NO: 373.
  • the multispecific antibody molecule comprises the anti-CDl 17 antibody molecule Antibody 48, or a derivative or clonal relative thereof.
  • the multispecific antibody molecule comprises a moiety that binds to the same, or substantially the same, epitope of an anti-CD3 antibody described herein and a
  • the multispecific antibody molecule competes for binding to CD3 with an anti-CD3 antibody described herein and competes for binding to CD117 with an antiCD 117 antibody described herein.
  • the multispecific antibody molecules described herein can further include a linker, e.g., a linker between the CD3 -targeting moiety and the CD117-targeting moiety.
  • the linker is chosen from: a cleavable linker, a non-cleavable linker, a peptide linker, a flexible linker, a rigid linker, a helical linker, or a non-helical linker, or a combination thereof.
  • the multispecific molecule can include one, two, three or four linkers, e.g., a peptide linker.
  • the peptide linker includes Gly and Ser.
  • the peptide linker comprises Gly and Ser.
  • the peptide linker is selected from GGGGS; GGGGSGGGGS; GGGGSGGGGSGGGGS; and DVPSGPGGGGGSGGGGS.
  • the peptide linker is a A(EAAAK)nA family of linkers (e.g., as described in Protein Eng. (2001) 14 (8): 529-532). These are stiff helical linkers with n ranging from 2 - 5.
  • the peptide linker is selected from AEAAAKEAAAKAAA; AEAAAKEAAAKEAAAKAAA; AEAAAKEAAAKEAAAKEAAAKAAA; and AEAAAKEAAAKEAAAKEAAAKEAAKAAA.
  • Nucleic acids encoding the antibody molecules (e.g., multispecific antibody molecules) described herein are also disclosed.
  • the invention features one or more nucleic acids comprising nucleotide sequences that encode heavy and light chain variable regions and CDRs or hypervariable loops of the antibody molecules, as described herein.
  • the invention features a first and second nucleic acid encoding heavy and light chain variable regions, respectively, of an antibody molecule chosen from one or more of the antibody molecules disclosed herein.
  • the one or more nucleic acids can each comprise a nucleotide sequence of an 163
  • the one or more nucleic acids can each comprise one or more nucleotide sequences encoding at least one, two, or three CDRs or hypervariable loops from a heavy chain variable region having an amino acid sequence of an antibody as set forth in the tables herein, or a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, and/or having one or more substitutions, e.g., conserved substitutions).
  • the one or more nucleic acids can each comprise a nucleotide sequence encoding at least one, two, or three CDRs or hypervariable loops from a light chain variable region having an amino acid sequence of an antibody as set forth in the tables herein, or a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, and/or having one or more substitutions, e.g., conserved substitutions).
  • the one or more nucleic acids can each comprise a nucleotide sequence encoding at least one, two, three, four, five, or six CDRs or hypervariable loops from heavy and light chain variable regions having an amino acid sequence of an antibody as set forth in the tables herein, or a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, and/or having one or more substitutions, e.g., conserved substitutions).
  • the one or more nucleic acids can each comprise a nucleotide sequence encoding at least one, two, or three CDRs or hypervariable loops from a heavy chain variable region having the nucleotide sequence of an antibody as set forth in the tables herein, a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, and/or capable of hybridizing under the stringency conditions described herein).
  • the one or more nucleic acids can each comprise a nucleotide sequence encoding at least one, two, or three CDRs or hypervariable loops from a light chain variable region having the nucleotide sequence of an antibody as set forth in the tables herein, or a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, and/or capable of hybridizing under the stringency conditions described herein).
  • the one or more nucleic 164 can each comprise a nucleotide sequence encoding at least one, two, or three CDRs or hypervariable loops from a light chain variable region having the nucleotide sequence of an antibody as set forth in the tables herein, or a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, and/or capable of hybridizing under the stringency conditions described herein).
  • T2110-7026WO acids can each comprise a nucleotide sequence encoding at least one, two, three, four, five, or six CDRs or hypervariable loops from heavy and light chain variable regions having the nucleotide sequence of an antibody as set forth in the tables herein, or a sequence substantially homologous thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, and/or capable of hybridizing under the stringency conditions described herein).
  • the one or more nucleic acids can each comprise a nucleotide sequence encoding a cytokine molecule (or a modulator of a cytokine molecule), an immune cell engager, or a stromal modifying moiety disclosed herein.
  • the application features one or more host cells and one or more vectors containing the one or more nucleic acids described herein.
  • the nucleic acids may be present in a single vector or separate vectors present in the same host cell or separate host cell, as described in more detail herein, below.
  • one or more vectors comprising the one or more nucleotide sequences encoding an antibody molecule (e.g., a multispecific antibody molecule) described herein.
  • the one or more vectors comprise one or more nucleotides encoding an antibody molecule (e.g., a multispecific antibody molecule) described herein.
  • the one or more vectors comprise the one or more nucleotide sequences described herein.
  • the one or more vectors include, but are not limited to, a virus, plasmid, cosmid, lambda phage, or a yeast artificial chromosome (YAC).
  • vectors utilize DNA elements which are derived from animal viruses such as, for example, bovine papilloma virus, polyoma virus, adenovirus, vaccinia virus, baculovirus, retroviruses (Rous Sarcoma Virus, MMTV or MOMLV) or SV40 virus.
  • DNA elements which are derived from animal viruses such as, for example, bovine papilloma virus, polyoma virus, adenovirus, vaccinia virus, baculovirus, retroviruses (Rous Sarcoma Virus, MMTV or MOMLV) or SV40 virus.
  • RNA elements derived from RNA viruses such as Semliki Forest virus, Eastern Equine Encephalitis virus and Flaviviruses.
  • cells which have stably integrated the DNA into their chromosomes may be selected by introducing one or more markers which allow for the selection of transfected host cells.
  • the marker may provide, for example, prototropy to an auxotrophic host, biocide resistance e.g., antibiotics), or resistance to heavy metals such as copper, or the like.
  • T2110-7026WO selectable marker gene can be either directly linked to the DNA sequences to be expressed or introduced into the same cell by cotransformation. Additional elements may also be needed for optimal synthesis of mRNA. These elements may include splice signals, as well as transcriptional promoters, enhancers, and termination signals.
  • the one or more expression vectors may be transfected or introduced into one or more appropriate host cells.
  • Various techniques may be employed to achieve this, such as, for example, protoplast fusion, calcium phosphate precipitation, electroporation, retroviral transduction, viral transfection, gene gun, lipid-based transfection or other conventional techniques.
  • protoplast fusion the cells are grown in media and screened for the appropriate activity. Methods and conditions for culturing the resulting transfected cells and for recovering the antibody molecule produced are known to those skilled in the art and may be varied or optimized depending upon the specific expression vector and mammalian host cell employed, based upon the present description.
  • the application features one or more host cells and one or more vectors containing the one or more nucleic acids described herein.
  • the nucleic acids may be present in a single vector or separate vectors present in the same host cell or separate host cell.
  • the one or more host cells can be one or more eukaryotic host cells, e.g., one or more mammalian cells, one or more insect cells, one or more yeast cells, or one or more prokaryotic cells, e.g., E. coli.
  • the one or more mammalian cells can be a cultured cell or a cell line.
  • Exemplary mammalian cells include lymphocytic cell lines (e.g., NSO), Chinese hamster ovary cells (CHO), COS cells, oocyte cells, and cells from a transgenic animal, e.g., mammary epithelial cells.
  • lymphocytic cell lines e.g., NSO
  • CHO Chinese hamster ovary cells
  • COS cells e.g., COS cells
  • oocyte cells e.g., oocyte cells
  • cells from a transgenic animal e.g., mammary epithelial cells.
  • the invention also provides one or more host cells comprising one or more nucleic acids encoding an antibody molecule as described herein.
  • the one or more host cells are genetically engineered to comprise one or more nucleic acids encoding the antibody molecule.
  • the one or more host cells are genetically engineered by using an expression cassette.
  • expression cassette refers to nucleotide sequences, which are capable of affecting expression of a gene in hosts compatible with such sequences.
  • T2110-7026WO cassettes may include a promoter, an open reading frame with or without introns, and a termination signal. Additional factors necessary or helpful in effecting expression may also be used, such as, for example, an inducible promoter.
  • the invention also provides one or more host cells comprising the one or more vectors described herein.
  • the one or more host cells can be, but are not limited to, one or more eukaryotic cells, one or more bacterial cells, one or more insect cells, or one or more human cells.
  • Suitable eukaryotic cells include, but are not limited to, Vero cells, HeLa cells, COS cells, CHO cells, HEK293 cells, BHK cells and MDCKII cells.
  • Suitable insect cells include, but are not limited to, Sf9 cells.
  • the antibody molecules (e.g., multispecific antibody molecules) described herein can be used in a method of treating a subject or a disorder.
  • Methods described herein include, for example, preparing a subject for hematopoietic stem cell transplantation or organ transplantation (/'. ⁇ ?., conditioning therapy) and treating a cancer in a subject by using a multispecific antibody molecule described herein, e.g., using a pharmaceutical composition described herein.
  • Hematopoietic stem cells are clonogenic, self-renewing pluripotent cells capable of ultimately differentiating into all cell types of the hematopoietic system, including B cells, T cells, NK cells, lymphoid dendritic cells, myeloid dendritic cells, granulocytes, macrophages, megakaryocytes, and erythroid cells.
  • the methods described herein reduce the likelihood of serious short-term and long-term consequences from standard of care pre-transplant conditioning therapy (e.g., myeloablative chemotherapy or radiation therapy).
  • the methods described herein (a) reduce anemia and thrombocytopenia; (b) reduce organ damage (e.g., veno-occlusive disease (VOD), renal failure, cardiac damage, or lung damage); (c) reduce sterility dysfunction; (d) reduce endocrine dysfunction; (e) reduce cognitive decline; (f) reduce secondary malignancies; (g) reduce mortality; (h) reduce infections; (i) reduce neutropenia;
  • organ damage e.g., veno-occlusive disease (VOD), renal failure, cardiac damage, or lung damage
  • the methods described herein reduce the likelihood of rejection of autologous stem cell transplantation (ASCT) or allogenic stem cell transplantation (allo-SCT) in a subject administered with a multispecific antibody molecule described herein, or a pharmaceutical composition described herein as compared to a subject not receiving the multispecific antibody molecule e.g., a subject receiving standard of care pre-transplantation conditioning therapy).
  • ASCT autologous stem cell transplantation
  • allo-SCT allogenic stem cell transplantation
  • allogenic refers to cells that are derived from, originate in, or are members of the same species, where the members are genetically related or genetically unrelated but genetically similar.
  • An “allogeneic transplant” refers to the transfer of cells from a donor to a recipient, where the recipient is the same species as the donor.
  • autologous refers to cells that are derived from, or originate in, the same subject or patient.
  • An “autologous transplant” refers to the harvesting and reinfusion or transplant of a subject’s own cells. Exclusive or supplemental use of autologous cells can eliminate or reduce many adverse effects of administration of the cells back to the host (e.g., graft versus host rejection).
  • the methods described herein reduce the likelihood of rejection of solid organ transplantation in a subject administered with a multispecific antibody molecule described herein, or a pharmaceutical composition described herein as compared to a subject not receiving the multispecific antibody molecule (e.g., a subject receiving standard of care pre-transplantation conditioning therapy).
  • a subject not receiving the multispecific antibody molecule e.g., a subject receiving standard of care pre-transplantation conditioning therapy.
  • the replacement of standard of care pre-transplantation toxic conditioning with a conditioning regimen comprising a multispecific molecule described herein followed by HSCT can, in some embodiments, induce host tolerance to solid organ transplants.
  • such a therapy may enable discontinuation of lifelong immunosuppressive agent regimens (e.g., rapamycin or cyclosporin) resulting in, for example, reduced rates of post transplantation infection and malignancy.
  • the subject has a primary, non-malignant hematological disorder.
  • non-malignant hematological disorder and “non-malignant hematological disease” refer to conditions affecting the hematopoietic and lymphoid tissues that are non-cancerous in 168
  • Hematopoietic and lymphoid tissues include the blood, bone marrow, lymph, and lymphatic system.
  • exemplary non-malignant hematological disorders include, but are not limited to, sickle cell disorder, thalassemia (e.g., beta thalassemia major), Fanconi anemia, amyloidosis, severe aplastic anemia, paroxysmal nocturnal hemoglobinuria, Hurler’s syndrome/mucopolysaccharidosis Type IH (MPS-IH), familial erythrophagocytic lymphohistiocytosis, and other histiocytic disorders,
  • the subject has an autoimmune disorder.
  • autoimmune disorders and “autoimmune diseases” refer to a variety of disorders in which a subject’s immune system produces antibodies directed against normal body tissues. Autoimmune diseases are characterized by the presence of self-antigens, immune system overactivity, and inflammation. Autoimmune diseases affect all major body systems including connective tissue, nerves, muscles, the endocrine system, skin, blood, and the respiratory and gastrointestinal systems.
  • autoimmune disorders include, but are not limited to, rheumatoid arthritis (RA), multiple sclerosis (MS), systemic lupus erythematosus (SLE), type I diabetes, Crohn’s disease, human immunodeficiency virus (HIV), autoimmune cytopenias, systemic sclerosis, chronic inflammatory demyelinating polyradiculoneuropathy, adrenoleukodystrophy (ALD), metachromatic leukodystrophy, severe combined immunodeficiency (SCID), and Wiskott- Aldrich syndrome.
  • RA rheumatoid arthritis
  • MS multiple sclerosis
  • SLE systemic lupus erythematosus
  • HAV human immunodeficiency virus
  • autoimmune cytopenias systemic sclerosis
  • chronic inflammatory demyelinating polyradiculoneuropathy adrenoleukodystrophy (ALD), metachromatic leukodystrophy, severe combined immunodeficiency
  • treating patients with severe autoimmune disorders e.g., MS, systemic sclerosis, or SLE
  • a conditioning regimen comprising a multispecific antibody molecule described herein and, for example, HSCT may serve to “reset the immune system” and ameliorate one or more symptoms of the disorder.
  • the methods described herein decrease the size of a tumor and/or decrease the number of cancer cells in a subject administered with a multispecific antibody molecule described herein, or a pharmaceutical composition described herein.
  • the methods described herein decrease tumor-based immunosuppression in a subject administered with a multispecific antibody molecule described herein, or a pharmaceutical composition described herein.
  • the cancer is a hematological cancer.
  • the hematological cancer is a leukemia or a lymphoma.
  • a “hematologic cancer” refers to a tumor of the hematopoietic or lymphoid tissues, e.g., a tumor that affects blood, bone marrow, or lymph nodes.
  • Exemplary hematologic malignancies include, but are not limited to, leukemia e.g., acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia (CML), hairy cell leukemia, acute monocytic leukemia (AMoL), chronic myelomonocytic leukemia (CMML), juvenile myelomonocytic leukemia (JMML), or large granular lymphocytic leukemia), lymphoma (e.g., AIDS-related lymphoma, cutaneous T-cell lymphoma, Hodgkin lymphoma (e.g., classical Hodgkin lymphoma or nodular lymphocyte-predominant Hodgkin lymphoma), mycosis fungoides, non-Hodgkin lymphoma (e.g., B-cell non-Hodgkin lymphoma (e
  • the cancer is a myeloproliferative neoplasm, e.g., primary or idiopathic myelofibrosis (MF), essential thrombocytosis (ET), polycythemia vera (PV), or chronic myelogenous leukemia (CML).
  • the cancer is myelofibrosis.
  • the subject has myelofibrosis.
  • the subject does not have the JAK2-V617F mutation.
  • the subject has the JAK2-V617F mutation.
  • the cancer is a solid cancer.
  • exemplary solid cancers include, but are not limited to, gastrointestinal cancers (e.g., gastrointestinal stromal tumor (GIST), ovarian cancer, rectal cancer, stomach cancer, testicular cancer, cancer of the anal region, uterine cancer, colon cancer, rectal cancer, renal-cell carcinoma, liver cancer, non- small cell carcinoma of the lung, cancer of the small intestine, cancer of the esophagus, melanoma, Kaposi’s sarcoma, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, 170
  • GIST gastrointestinal stromal tumor
  • ovarian cancer rectal cancer
  • rectal cancer stomach cancer
  • testicular cancer cancer of the anal region
  • colon cancer rectal cancer
  • renal-cell carcinoma liver cancer
  • non- small cell carcinoma of the lung cancer of the small intestine
  • Exemplary cancers characterized by immune suppression that may be treated according to the methods herein include, but are not limited to, GIST and AML. Further exemplary cancers that may be treated according to the methods herein include, but are not limited to, seminomas, malignant melanomas, adenoid-cystic sarcomas, and large-cell carcinomas of the lung.
  • the cancer has a mutation in CD117. In some embodiments, the mutation is a gain of function mutation.
  • the multispecific antibody molecules are administered in a manner appropriate to the disease to be treated or prevented.
  • the quantity and frequency of administration will be determined by such factors as the condition of the patient, and the type and severity of the patient’s disease. Appropriate dosages may be determined by clinical trials. For example, when “an effective amount” or “a therapeutic amount” is indicated, the precise amount of the pharmaceutical composition (or multispecific antibody molecules) to be administered can be determined by a physician with consideration of individual differences in tumor size, extent of infection or metastasis, age, weight, and condition of the subject.
  • the pharmaceutical composition described herein can be administered at a dosage of about 0.01 mg/kg to about 100 mg/kg of patient body weight, including all integer values within that range.
  • the pharmaceutical compositions described herein may also be administered in a flat dosage of between about 100 mg to about 2000 mg, including all integer values within that range.
  • the pharmaceutical compositions described herein can be administered multiple times at these dosages.
  • the multispecific antibody molecules or pharmaceutical composition is administered to the subject parenterally. In some embodiments, the multispecific 171
  • T2110-7026WO antibody molecules or pharmaceutical compositions are administered to the subject intravenously, subcutaneously, intratumorally, intranodally, intramuscularly, intradermally, or intraperitoneally.
  • the multispecific antibody are administered, e.g., injected, directly into a tumor or lymph node.
  • the multispecific antibodycells are administered as an infusion (e.g., as described in Rosenberg et al., New Eng. J. of Med. 319:1676, 1988) or an intravenous push.
  • the multispecific antibody molecules or pharmaceutical compositions are administered as an injectable depot formulation.
  • the subject is a mammal. In some embodiments, the subject is a human, monkey, pig, dog, cat, cow, sheep, goat, rabbit, rat, or mouse. In an embodiment, the subject is a human. In some embodiments, the subject is a pediatric subject, e.g., less than 18 years of age, e.g., less than 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or less years of age. In some embodiments, the subject is an adult, e.g., at least 18 years of age, e.g., at least 19, 20, 21, 22, 23, 24, 25, 25-30, 30-35, 35-40, 40-50, 50-60, 60-70, 70-80, or 80-90 years of age.
  • the subject is a mammal. In some embodiments, the subject is a human, monkey, pig, dog, cat, cow, sheep, goat, rabbit, rat, or mouse. In an embodiment, the subject is a human. In some embodiments, the subject is a pediatric subject, e
  • the multispecific antibody molecules disclosed herein can be used in combination with a second therapeutic agent or procedure.
  • the multispecific antibody molecule described herein is administered in combination one or more additional therapeutic agents to increase tolerability of the antibody molecule.
  • the one or more additional therapeutic agents may decrease the frequency or duration of, or lessen the severity of hypersensitivity reactions and/or cytokine release syndrome (CRS).
  • cytokine release syndrome CRS
  • Exemplary therapeutic agents that may be used to improve tolerability of a multispecific molecule disclosed herein include, but are not limited to, antihistamines, acetaminophen, steroids (e.g., corticosteroids), and tocilizumab.
  • the second therapeutic agent is administered prior to, concurrently with, or after the multispecific antibody molecule described herein.
  • the multispecific antibody molecule and the second therapeutic agent or procedure are administered/performed after a subject has been diagnosed with a non- malignant hematological disorder or an autoimmune disorder. In some embodiments, the multispecific antibody molecule and the second therapeutic agent or procedure are administered/performed sequentially. For example, the delivery of one treatment ceases before the delivery of the other treatment begins.
  • combination therapy can lead to more effective treatment than monotherapy with either agent alone.
  • the combination of the first and second treatment is more effective (e.g., leads to a greater reduction in symptoms or complications (e.g., transplant rejection)) than the first or second treatment alone.
  • the combination therapy permits use of a lower dose of the first or the second treatment compared to the dose of the first or second treatment normally required to achieve similar effects when administered as a monotherapy.
  • the combination therapy has a partially additive effect, wholly additive effect, or greater than additive effect.
  • the multispecific antibody molecule is administered prior to (e.g., as a conditioning therapy prior to stem cell transplantation) administration of the second therapeutic agent or procedure.
  • the second therapeutic agent includes one or more of (a) radiation; (b) low dose busulfan; and (c) mobilization of HSC (e.g., using G-CSF, plerixafor (e.g., AMD3100), stem cell factor, or GROBT).
  • the second therapeutic procedure is HSCT.
  • the HSCT is allogenic HSCT.
  • the HSCT is autologous HSCT.
  • Transplantation of hematopoietic stem cells into an appropriate host may be accomplished by any method known to those of skill in the art.
  • intravenous infusion may be used.
  • the pharmaceutical composition described herein can be administered using infusion techniques described in immunotherapy (see, e.g., Rosenberg et al., New Eng. J. of Med. 319:1676, 1988).
  • the cells administered to the subject parenterally.
  • the cells are administered to the subject intravenously, subcutaneously, intratumorally, intranodally, intramuscularly, intradermally, or intraperitoneally.
  • the cells are administered, e.g., injected, directly into a tumor or lymph node.
  • the cells are administered as an infusion (e.g., as 173
  • the cells are administered as an injectable depot formulation.
  • the number of cells transfused will be dependent on certain factors including sex, age, weight, the types of disease or disorder, stage of the disorder, the percentage of the desired cells in the cell population (e.g., purity of cell population), and the cell number needed to produce a therapeutic benefit.
  • the numbers of expanded cells infused may be from about lxl0 4 to about lxl0 5 cells/kg, from about lxl0 5 to about 10xl0 6 cells/kg, preferably about IxlO 6 cells to about 5xl0 6 cells/kg of body weight, or more as necessary.
  • the cells are in a pharmaceutically acceptable carrier at about IxlO 9 to about 5xl0 9 cells.
  • Cells may be administered in one infusion, or through successive infusions over a defined time period sufficient to generate a therapeutic effect. Different populations of cells may be infused when treatment involves successive infusions.
  • a pharmaceutically acceptable carrier may be used for infusion of the cells into the patient.
  • the multispecific antibody molecule and the second therapeutic agent or procedure are administered/performed after a subject has been diagnosed with an autoimmune disorder or one who is in need of a solid organ transplantation. In some embodiments, the multispecific antibody molecule and the second therapeutic agent or procedure are administered/performed sequentially. For example, the delivery of one treatment ceases before the delivery of the other treatment begins.
  • combination therapy can lead to more effective treatment than monotherapy with either agent alone.
  • the combination of the first and second treatment is more effective (e.g., leads to a greater reduction in symptoms or complications (e.g., transplant rejection)) than the first or second treatment alone.
  • the combination therapy permits use of a lower dose of the first or the second treatment compared to the dose of the first or second treatment normally required to achieve similar effects when administered as a monotherapy.
  • the combination therapy has a partially additive effect, wholly additive effect, or greater than additive effect.
  • the multispecific antibody molecule is administered prior to administration of the second therapeutic agent or procedure.
  • the second 174 is administered prior to administration of the second therapeutic agent or procedure.
  • T2110-7026WO therapeutic agent includes one or more of cyclophosphamide, anti-thymocyte globulin (ATG), and alemtuzumab.
  • the multispecific antibody molecule and the second therapeutic agent or procedure are administered/performed after a subject has been diagnosed with a cancer, e.g., before the cancer has been eliminated from the subject.
  • the multispecific antibody molecule and the second therapeutic agent or procedure are administered/performed simultaneously or concurrently. For example, the delivery of one treatment is still occurring when the delivery of the second commences, e.g., there is an overlap in administration of the treatments.
  • the multispecific antibody molecule and the second therapeutic agent or procedure are administered/performed sequentially. For example, the delivery of one treatment ceases before the delivery of the other treatment begins.
  • combination therapy can lead to more effective treatment than monotherapy with either agent alone.
  • the combination of the first and second treatment is more effective (e.g., leads to a greater reduction in symptoms and/or cancer cells) than the first or second treatment alone.
  • the combination therapy permits use of a lower dose of the first or the second treatment compared to the dose of the first or second treatment normally required to achieve similar effects when administered as a monotherapy.
  • the combination therapy has a partially additive effect, wholly additive effect, or greater than additive effect.
  • the multispecific antibody molecule is administered in combination with a therapy, e.g., a cancer therapy (e.g., one or more of anti-cancer agents, immunotherapy, photodynamic therapy (PDT), surgery and/or radiation).
  • a cancer therapy e.g., one or more of anti-cancer agents, immunotherapy, photodynamic therapy (PDT), surgery and/or radiation.
  • chemotherapeutic chemotherapeutic agent
  • anti-cancer agent are used interchangeably herein.
  • the administration of the multispecific antibody molecule and the therapy e.g., the cancer therapy, can be sequential (with or without overlap) or simultaneous. Administration of the multispecific antibody molecule can be continuous or intermittent during the course of therapy (e.g., cancer therapy).
  • Certain therapies described herein can be used to treat cancers and non-cancerous diseases. For example, PDT efficacy can be enhanced in cancerous and non-cancerous
  • the multispecific antibody molecule is administered in combination with a low or small molecular weight chemotherapeutic agent.
  • chemotherapeutic agents include, but not limited to, 13-cis-retinoic acid (isotretinoin, ACCUTANE®), 2-CdA (2-chlorodeoxyadenosine, cladribine, LEUSTATINTM), 5- azacitidine (azacitidine, VID AZA®), 5 -fluorouracil (5-FU, fluorouracil, ADRUCIL®), 6- mercaptopurine (6-MP, mercaptopurine, PURINETHOL®), 6-TG (6-thioguanine, thioguanine, THIOGUANINE TABLOID®), abraxane (paclitaxel protein-bound), actinomycin-D (dactinomycin, COSMEGEN®), alitretinoin (PANRETIN®), all-transretinoic acid
  • the multispecific antibody molecule is administered in conjunction with a biologic.
  • Biologies useful in the treatment of cancers are known in the art and a binding molecule of the invention may be administered, for example, in conjunction with such known biologies.
  • the FDA has approved the following biologies for the treatment of breast cancer: HERCEPTIN® (trastuzumab, Genentech Inc., South San Francisco, Calif.; a humanized monoclonal antibody that has anti-tumor activity in HER2 -positive breast cancer); FASLODEX® (fulvestrant, AstraZeneca Pharmaceuticals, LP, Wilmington, Del.; an estrogen- receptor antagonist used to treat breast cancer); ARIMIDEX® (anastrozole, AstraZeneca Pharmaceuticals, LP; a nonsteroidal aromatase inhibitor which blocks aromatase, an enzyme needed to make estrogen); Aromasin® (exemestane, Pfizer Inc., New York, N.Y.; an irreversible, steroidal aromat
  • AVASTIN® bevacizumab, Genentech Inc.; the first FDA-approved therapy designed to inhibit angiogenesis
  • ZEVALIN® ibritumomab tiuxetan, Biogen Stahl, Cambridge, Mass.; a radiolabeled monoclonal antibody currently approved for the treatment of B-cell lymphomas
  • AVASTIN® AVASTIN®
  • ERBITUX® cetuximab, ImClone Systems Inc., New York, N.Y., and Bristol-Myers Squibb, New York, N.Y.
  • GLEEVEC® imatinib mesylate; a protein kinase inhibitor
  • exemplary biologies include TARCEVA® (erlotinib HCL, OSI Pharmaceuticals Inc., Melville, N.Y.; a small molecule designed to target the human epidermal growth factor receptor 1 (HER1) pathway).
  • TARCEVA® erlotinib HCL, OSI Pharmaceuticals Inc., Melville, N.Y.
  • HER1 human epidermal growth factor receptor 1
  • exemplary biologies include VELCADE® Velcade (bortezomib, Millennium Pharmaceuticals, Cambridge Mass.; a proteasome inhibitor). Additional biologies include THALIDOMID® (thalidomide, Clegene Corporation, Warren, N.J.; an immunomodulatory agent and appears to have multiple actions, including the ability to inhibit the growth and survival of myeloma cells and anti-angiogenesis).
  • Additional exemplary cancer therapeutic antibodies include, but are not limited to, 3F8, abagovomab, adecatumumab, afutuzumab, alacizumab pegol, alemtuzumab (CAMPATH®, MABCAMPATH®), altumomab pentetate (HYBRI-CEAKER®), anatumomab mafenatox, anrukinzumab (IMA-638), apolizumab, arcitumomab (CEA-SCAN®), bavituximab, bectumomab (LYMPHOSCAN®), belimumab (BENLYSTA®, LYMPHOSTAT-B®), besilesomab (SCINTIMUN®), bevacizumab (AVASTIN®), bivatuzumab mertansine, blinatumomab, brentuximab vedotin, cantuzumab mer
  • VECTIBIX® pemtumomab
  • THERAGYN® pertuzumab
  • OPTG® pintumomab
  • pritumumab pritumumab
  • ramucirumab ranibizumab
  • MABTHERA® rituximab
  • RITUXAN® ranibizumab
  • satumomab pendetide sibrotuzumab, siltuximab
  • AFP-CIDE® ticilimumab
  • tigatuzumab TNX-650
  • tositumomab HERCEPTIN®
  • tremelimumab tigatuzumab
  • TNX-650 TNX-650
  • tositumomab BEXXAR®
  • trastuzumab HERCEPTIN®
  • tremelimumab ticilimumab
  • the multispecific antibody molecule is administered in combination with a viral cancer therapeutic agent.
  • viral cancer therapeutic agents include, but not limited to, vaccinia virus (vvDD-CDSR), carcinoembryonic antigen-expressing measles virus, recombinant vaccinia virus (TK-deletion plus GM-CSF), Seneca Valley virus- 001, Newcastle virus, coxsackie virus A21, GL-ONC1, EBNA1 C-terminal/LMP2 chimeric protein-expressing recombinant modified vaccinia Ankara vaccine, carcinoembryonic antigenexpressing measles virus, G207 oncolytic virus, modified vaccinia virus Ankara vaccine expressing p53, OncoVEX GM-CSF modified herpes-simplex 1 virus, fowlpox virus vaccine vector, recombinant vaccinia pro state- specific antigen vaccine, human papillomavirus 16/18 LI virus-like particle/AS04 vaccine, MVA
  • L1/AS04 fowlpox virus vaccine vector, vaccinia-tyrosinase vaccine, MEDI-517 HPV-16/18 VLP AS04 vaccine, adenoviral vector containing the thymidine kinase of herpes simplex virus TK99UN, HspE7, FP253/Fludarabine, ALVAC(2) melanoma multi-antigen therapeutic vaccine, ALVAC-hB7.1, canarypox-hIL-12 melanoma vaccine, Ad-REIC/Dkk-3, rAd-IFN SCH 721015, TIL-Ad-INFg, Ad-ISF35, and coxsackievirus A21 (CVA21, CAVATAK®).
  • the multispecific antibody molecule is administered in combination with a nanopharmaceutical.
  • exemplary cancer nanopharmaceuticals include, but not limited to, ABRAXANE® (paclitaxel bound albumin nanoparticles), CRLX101 (CPT conjugated to a linear cyclodextrin-based polymer), CRLX288 (conjugating docetaxel to the biodegradable polymer poly (lactic-co-glycolic acid)), cytarabine liposomal (liposomal Ara-C, DEPOCYTTM), daunorubicin liposomal (DAUNOXOME®), doxorubicin liposomal (DOXIL®, CAELYX®), encapsulated-daunorubicin citrate liposome (DAUNOXOME®), and PEG anti- VEGF aptamer (MACUGEN®).
  • ABRAXANE® paclitaxel bound albumin nanoparticles
  • CRLX101 CPT
  • the multispecific antibody molecule is administered in combination with paclitaxel or a paclitaxel formulation, e.g., TAXOL®, protein-bound paclitaxel (e.g., ABRAXANE®).
  • a paclitaxel formulation e.g., TAXOL®, protein-bound paclitaxel (e.g., ABRAXANE®).
  • Exemplary paclitaxel formulations include, but are not limited to, nanoparticle albumin-bound paclitaxel (ABRAXANE®, marketed by Abraxis Bioscience), docosahexaenoic acid bound-paclitaxel (DHA-paclitaxel, Taxoprexin, marketed by Protarga), polyglutamate bound-paclitaxel (PG-paclitaxel, paclitaxel poliglumex, CT-2103, XYOTAX, marketed by Cell Therapeutic), the tumor-activated prodrug (TAP), ANG105 (Angiopep-2 bound to three molecules of paclitaxel, marketed by ImmunoGen), paclitaxel-EC-1 (paclitaxel bound to the erbB2-recognizing peptide EC-1; see Li et al., Biopolymers (2007) 87:225-230), and glucose-conjugated paclitaxel (e.g., 2’ -paclitaxel methyl 2-glucopyrano
  • RNAi and antisense RNA agents for treating cancer include, but not limited to, CALAA-01, siG12D LODER (Local Drug EluteR), and ALN-VSP02.
  • cancer therapeutic agents include, but not limited to, cytokines (e.g., aldesleukin (IL-2, Interleukin-2, PROLEUKIN®), alpha Interferon (IFN-alpha, Interferon alfa, INTRON® A (Interferon alfa- 2b), ROFERON-A® (Interferon alfa-2a)), Epoetin alfa (PROCRIT®), filgrastim (G-CSF, Granulocyte - Colony Stimulating Factor, NEUPOGEN®), GM-CSF (Granulocyte Macrophage Colony Stimulating Factor, sargramostim, LEUKINETM), IL- 11 (Interleukin-11, 180
  • cytokines e.g., aldesleukin (IL-2, Interleukin-2, PROLEUKIN®
  • IFN-alpha Interferon alfa
  • INTRON® A Interferon alfa- 2b
  • the multispecific antibody molecule is used in combination with a tyrosine kinase inhibitor (e.g., a receptor tyrosine kinase (RTK) inhibitor).
  • a tyrosine kinase inhibitor e.g., a receptor tyrosine kinase (RTK) inhibitor.
  • exemplary tyrosine kinase inhibitor include, but are not limited to, an epidermal growth factor (EGF) pathway inhibitor (e.g., an epidermal growth factor receptor (EGFR) inhibitor), a vascular endothelial growth factor (VEGF) pathway inhibitor (e.g., an antibody against VEGF, a VEGF trap, a vascular endothelial growth factor receptor (VEGFR) inhibitor (e.g., a VEGFR-1 inhibitor, a 181
  • EGF epidermal growth factor
  • VEGF vascular endothelial growth factor
  • VEGFR-1 inhibitor e.g., a
  • VEGFR-2 inhibitor a VEGFR-3 inhibitor
  • PDGF platelet derived growth factor pathway inhibitor
  • PDGFR platelet derived growth factor receptor
  • RAF-1 inhibitor a KIT inhibitor and a RET inhibitor.
  • the anti-cancer agent used in combination with the AHCM agent is selected from the group consisting of: axitinib (AG013736), bosutinib (SKI-606), cediranib (RECENTINTM, AZD2171), dasatinib (SPRYCEL®, BMS-354825), erlotinib (TARCEVA®), gefitinib (IRESSA®), imatinib (Gleevec®, CGP57148B, STI-571), lapatinib (TYKERB®, TYVERB®), lestaurtinib (CEP-701), neratinib (HKI-272), nilotinib (TASIGNA®), semaxanib (semaxinib, SU5416), sunitinib (SUTENT®, SU11248), toceranib (PALLADIA®), vandetanib (ZACTIMA®, ZD6474), vatalanib (AG01
  • tyrosine kinase inhibitors are chosen from sunitinib, erlotinib, gefitinib, or sorafenib. In one embodiment, the tyrosine kinase inhibitor is sunitinib.
  • the multispecific antibody molecule is administered in combination with one of more of: an anti- angiogenic agent, or a vascular targeting agent or a vascular disrupting agent.
  • anti- angiogenic agents include, but are not limited to, VEGF inhibitors (e.g., anti- VEGF antibodies (e.g., bevacizumab); VEGF receptor inhibitors (e.g., itraconazole); inhibitors of cell proliferatin and/or migration of endothelial cells (e.g., carboxyamidotriazole, TNP-470); inhibitors of angiogenesis stimulators (e.g., suramin), among 182
  • VEGF inhibitors e.g., anti- VEGF antibodies (e.g., bevacizumab); VEGF receptor inhibitors (e.g., itraconazole); inhibitors of cell proliferatin and/or migration of endothelial cells (e.g., carboxyamidotriazole, TNP-470
  • VTA vascular-targeting agent
  • VDA vascular disrupting agent
  • a vascular-targeting agent or VDA is designed to damage the vasculature (blood vessels) of cancer tumors causing central necrosis (reviewed in, e.g., Thorpe, P.E. (2004) Clin. Cancer Res. Vol. 10:415-427).
  • VTAs can be small-molecule.
  • Exemplary small-molecule VTAs include, but are not limited to, microtubule destabilizing drugs (e.g., combretastatin A-4 disodium phosphate (CA4P), ZD6126, AVE8062, Oxi 4503); and vadimezan (ASA404).
  • the second therapeutic agent comprises cytarabine and/or daunorubicin.
  • the second therapeutic agent comprises azacytidine and/or decitabine.
  • methods described herein comprise use of an immune checkpoint inhibitor in combination with the multispecific antibody molecule.
  • the methods can be used in a therapeutic protocol in vivo.
  • an immune checkpoint inhibitor inhibits a checkpoint molecule.
  • checkpoint molecules include but are not limited to CTLA4, PD1, PD-L1, PD-L2, TIM3, LAG3, CD160, 2B4, CD80, CD86, B7-H3 (CD276), B7-H4 (VTCN1), HVEM (TNFRSF14 or CD270), BTLA, KIR, MHC class I, MHC class II, GAL9, VISTA, BTLA, TIGIT, LAIR1, and A2aR. See, e.g., Pardoll. Nat. Rev. Cancer 12.4(2012):252-64, incorporated herein by reference.
  • the immune checkpoint inhibitor is a PD-1 inhibitor, e.g., an anti- PD-1 antibody such as Nivolumab, Pembrolizumab or Pidilizumab.
  • Nivolumab also called MDX- 1106, MDX-1106-04, ONO-4538, or BMS-936558
  • Pembrolizumab (also called Lambrolizumab, MK-3475, MK03475, SCH-900475 or KEYTRUDA®; Merck) is a humanized IgG4 monoclonal antibody that binds to PD-1. See, e.g., Hamid, O. et al. (2013) New England. Journal of Medicine 369 (2): 134-44, US 8,354,509 and W02009/114335.
  • Pidilizumab (also called CT-011 or Cure Tech) is a humanized IgGlk monoclonal antibody that binds to PD1. See, e.g., W02009/101611.
  • the inhibitor of PD-1 is an antibody molecule having a sequence substantially identical or similar thereto, e.g., a sequence at least 85%, 90%, 95% identical or higher to the sequence of
  • Nivolumab Nivolumab, Pembrolizumab or Pidilizumab. Additional anti-PDl antibodies, e.g., AMP 514 (Amplimmune), are described, e.g., in US 8,609,089, US 2010028330, and/or US 20120114649.
  • AMP 514 Amplimmune
  • the PD-1 inhibitor is an immunoadhesin, e.g., an immunoadhesin comprising an extracellular/PD- 1 binding portion of a PD-1 ligand (e.g., PD-L1 or PD-L2) that is fused to a constant region (e.g., an Fc region of an immunoglobulin).
  • a PD-1 ligand e.g., PD-L1 or PD-L2
  • a constant region e.g., an Fc region of an immunoglobulin.
  • the PD-1 inhibitor is AMP-224 (B7-DCIg, e.g., described in WO2011/066342and W02010/027827), a PD-L2 Fc fusion soluble receptor that blocks the interaction between B7-H1 and PD-1.
  • the immune checkpoint inhibitor is a PD-L1 inhibitor, e.g., an antibody molecule.
  • the PD-L1 inhibitor is YW243.55.S70, MPDL3280A, MEDI-4736, MSB-0010718C, or MDX-1105.
  • the anti- PD-L1 antibody is MSB0010718C (also called A09-246-2; Merck Serono), which is a monoclonal antibody that binds to PD-L1.
  • Exemplary humanized anti-PD-Ll antibodies are described, e.g., in WO2013/079174.
  • the PD-L1 inhibitor is an anti-PD-Ll antibody, e.g., YW243.55.S70.
  • the YW243.55.S70 antibody is described, e.g., in WO 2010/077634.
  • the PD-L1 inhibitor is MDX-1105 (also called BMS- 936559), which is described, e.g., in W02007/005874.
  • the PD-L1 inhibitor is MDPL3280A (Genentech / Roche), which is a human Fc-optimized IgGl monoclonal antibody against PD-L1. See, e.g., U.S.
  • the inhibitor of PD-L1 is an antibody molecule having a sequence substantially identical or similar thereto, e.g., a sequence at least 85%, 90%, 95% identical or higher to the sequence of YW243.55.S70, MPDL3280A, MEDI-4736, MSB- 0010718C, or MDX-1105.
  • the immune checkpoint inhibitor is a PD-L2 inhibitor, e.g., AMP- 224 (which is a PD-L2 Fc fusion soluble receptor that blocks the interaction between PD1 and B7-H1. See, e.g., W02010/027827 and WO2011/066342.
  • AMP- 224 which is a PD-L2 Fc fusion soluble receptor that blocks the interaction between PD1 and B7-H1. See, e.g., W02010/027827 and WO2011/066342.
  • the immune checkpoint inhibitor is a LAG-3 inhibitor, e.g., an anti LAG-3 antibody molecule.
  • the anti-LAG-3 antibody is BMS-986016 (also called BMS986016; Bristol-Myers Squibb). BMS-986016 and other humanized anti-LAG- 3 antibodies are described, e.g., in US 2011/0150892, W02010/019570, and W02014/008218.
  • the immune checkpoint inhibitor is a TIM-3 inhibitor, e.g., anti- TIM3 antibody molecule, e.g., described in U.S. Patent No.: 8,552,156, WO 2011/155607, EP 2581113 and U.S Publication No.: 2014/044728.
  • a TIM-3 inhibitor e.g., anti- TIM3 antibody molecule, e.g., described in U.S. Patent No.: 8,552,156, WO 2011/155607, EP 2581113 and U.S Publication No.: 2014/044728.
  • the immune checkpoint inhibitor is a CTLA-4 inhibitor, e.g., anti- CTLA-4 antibody molecule.
  • CTLA-4 inhibitor e.g., anti- CTLA-4 antibody molecule.
  • Exemplary anti-CTLA4 antibodies include Tremelimumab (IgG2 monoclonal antibody from Pfizer, formerly known as ticilimumab, CP-675,206); and Ipilimumab (also called MDX-010, CAS No. 477202-00-9).
  • Tremelimumab IgG2 monoclonal antibody from Pfizer, formerly known as ticilimumab, CP-675,206
  • Ipilimumab also called MDX-010, CAS No. 477202-00-9
  • Other exemplary anti-CTLA-4 antibodies are described, e.g., in U.S. Pat. No. 5,811,097.
  • a multispecific antibody molecule comprising a first moiety that binds to CD3; and a second moiety that binds to CD117.
  • the multispecific antibody molecule of embodiment 1 or 2, wherein the first moiety that binds to CD3 comprises:
  • HCDR1, HCDR2, or HCDR3 of an anti-CD3 antibody described herein or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions;
  • VL of an anti-CD3 antibody described herein or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions; or
  • CD117 is a hematopoietic stem cell (HSC), a multipotent progenitor (MPP), a common myeloid progenitor (CMP), or a cancer cell (e.g., a GIST cancer cell or an AML cancer cell).
  • HSC hematopoietic stem cell
  • MPP multipotent progenitor
  • CMP common myeloid progenitor
  • cancer cell e.g., a GIST cancer cell or an AML cancer cell.
  • an immunoglobulin chain constant region e.g., Fc region
  • the mutation comprises one or more substitution mutations at one or more of amino acid residues selected from the group consisting of Ile253, Ser254, His435, Tyr436, Glu233-Gly236, Arg255, Lys288, Ser415, His310, and His433.
  • the dimerization module comprises an immunoglobulin constant domain, e.g., a heavy chain constant domain (e.g., a homodimeric or heterodimeric heavy chain constant region, e.g., an Fc region), or a constant domain of an immunoglobulin variable region (e.g., a Fab region).
  • a heavy chain constant domain e.g., a homodimeric or heterodimeric heavy chain constant region, e.g., an Fc region
  • a constant domain of an immunoglobulin variable region e.g., a Fab region
  • the dimerization module comprises one or more immunoglobulin chain constant regions (e.g., Fc regions) comprising one or more of: a paired cavity-protuberance (“knob-in-a hole”), an electrostatic interaction, or a strand-exchange.
  • immunoglobulin chain constant regions e.g., Fc regions
  • the one or more immunoglobulin chain constant regions comprise an amino acid substitution at a position chosen from one or more of 347, 349, 350, 351, 366, 368, 370, 392, 394, 395, 397, 398, 399, 405, 407, or 409, e.g., of the Fc region of human IgGl, optionally wherein the one or more immunoglobulin chain constant regions (e.g., Fc regions) comprise an amino acid substitution chosen from: T366S, L368A, or Y407V (e.g., corresponding to a cavity or hole), or T366W (e.g., corresponding to a protuberance or knob), or a combination thereof.
  • the one or more immunoglobulin chain constant regions comprise an amino acid substitution at a position chosen from one or more of 347, 349, 350, 351, 366, 368, 370, 392, 394, 395, 397, 398, 399, 405, 407, or 409,
  • a linker e.g., a linker between the first moiety that binds to CD3 and the second moiety that binds to CD117.
  • linker is chosen from: a cleavable linker, a non-cleavable linker, a peptide linker, a flexible linker, a rigid linker, a helical linker, or a non-helical linker.
  • HCDR1, HCDR2, or HCDR3 of an anti-CD3 antibody is selected from a sequence described in Table 2C herein;
  • HCDR1, HCDR2, or HCDR3 of an anti-CDl 17 antibody is selected from a sequence described in Table 3C herein;
  • a vector e.g., an expression vector, comprising a nucleic acid of embodiment 33.
  • a cell e.g., a host cell, comprising the nucleic acid of embodiment 33 or the vector of claim 34.
  • a composition e.g., a pharmaceutical composition, comprising the multispecific antibody molecule of any of embodiments 1-32, optionally further comprising a pharmaceutically acceptable carrier, excipient, or stabilizer.
  • kits comprising the multispecific antibody molecule of any of embodiments 1-32, and instructions to use the multispecific antibody molecule.
  • a method of preparing a subject for a hematopoietic stem cell transplantation comprising administering to a subject in need thereof an effective amount of the multispecific antibody molecule of any of embodiments 1-33 or the composition of claim 37, thereby preparing the subject.
  • HSCT hematopoietic stem cell transplantation
  • autoimmune disorder is lupus, rheumatoid, arthritis (RA), multiple sclerosis (MS), type I diabetes, or Crohn’s disease.
  • a method of treating a disorder in a subject comprising administering to the subject an effective amount of the multispecific antibody molecule of any of embodiments 1-32 or the composition of embodiment 37, wherein the subject is in need of receiving an HSCT, thereby treating the disorder.
  • a method of treating a disorder comprising administering to the subject in need thereof an effective amount of the multispecific antibody molecule of any of embodiments 1-32 or the pharmaceutical composition of embodiment 37, and an HSCT, thereby treating the disorder.
  • any one of embodiments 39-50 wherein the subject exhibits one or more of (a) reduced anemia and thrombocytopenia; (b) reduced organ damage; (c) reduced sterility; (d) reduced endocrine dysfunction; (e) reduced cognitive decline; (f) reduced secondary malignancies; (g) reduced mortality; (h) reduced infections; (i) reduced neutropenia; and/or (j) reduced reactivation of latent viruses (e.g., EBV and CMV) as compared to a subject receiving myeloablative chemotherapy or radiation therapy.
  • latent viruses e.g., EBV and CMV
  • a method of reducing immunosuppression comprising administering to a subject in need thereof an effective amount of the multispecific antibody molecule of any of claims 1-32 or the pharmaceutical composition of embodiment 37, thereby reducing immunosuppression.
  • a method of treating a cancer comprising administering to a subject in need thereof an effective amount of the multispecific antibody molecule of any one of claims 1-32 or the pharmaceutical composition of embodiment 37, thereby treating the cancer.
  • the cancer is a solid tumor cancer.
  • the cancer is an adenoid-cystic sarcoma, a seminoma, a malignant melanoma, an adenoid-cystic carcinoma, or a large-cell carcinoma of the lung.
  • the second therapy comprises a chemotherapy, a radiation therapy, a surgery, a targeted therapy, or an immunotherapy.
  • anti-KIT hybridoma supernatants were harvested and used to stain different KIT expressing cell lines and analyzed by flow cytometry. Briefly, the cells were incubated in serially diluted supernatant for 45 minutes, washed, and subsequently incubated in FITC-labeled goat anti-mouse secondary, washed again, and then measured using a Fortessa flow cytometer.
  • Table A shows the mean fluorescence intensity of a 4-fold dilution series on human KIT- expressing 293 cells.
  • Table B shows the mean fluorescence intensity of a 4 or 5-fold dilution series on cyno KIT- expressing 293 cells.

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La divulgation concerne des molécules d'anticorps multispécifiques qui comprennent une première fraction qui se lie à CD3 et une seconde fraction qui se lie à CD117.
PCT/US2021/063558 2020-12-16 2021-12-15 Molécules d'anticorps multispécifiques et leurs utilisations WO2022132929A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202063126311P 2020-12-16 2020-12-16
US63/126,311 2020-12-16

Publications (2)

Publication Number Publication Date
WO2022132929A2 true WO2022132929A2 (fr) 2022-06-23
WO2022132929A3 WO2022132929A3 (fr) 2022-09-01

Family

ID=82060095

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2021/063558 WO2022132929A2 (fr) 2020-12-16 2021-12-15 Molécules d'anticorps multispécifiques et leurs utilisations

Country Status (1)

Country Link
WO (1) WO2022132929A2 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12006366B2 (en) 2020-06-11 2024-06-11 Provention Bio, Inc. Methods and compositions for preventing type 1 diabetes

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008118324A2 (fr) * 2007-03-26 2008-10-02 Macrogenics, Inc. Composition et procédé de traitement du cancer avec un anticorps anti-uroplakine ib
EP3105252B1 (fr) * 2014-02-12 2019-07-24 Michael Uhlin Anticorps bispécifiques utilisables dans une transplantation de cellules souches
CA2949033A1 (fr) * 2015-11-30 2017-05-30 Pfizer Inc. Anticorps et fragments d'anticorps destines a la conjugaison specifique au site
SG10201913823VA (en) * 2016-10-07 2020-03-30 Novartis Ag Chimeric antigen receptors for the treatment of cancer
AU2017378451A1 (en) * 2016-12-15 2019-07-04 The National Institute for Biotechnology in the Negev Ltd. Anti-PCNA monoclonal antibodies and use thereof
WO2020219775A1 (fr) * 2019-04-24 2020-10-29 Magenta Therapeutics, Inc. Conjugués anticorps-médicament anti-cd117 et leurs utilisations

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12006366B2 (en) 2020-06-11 2024-06-11 Provention Bio, Inc. Methods and compositions for preventing type 1 diabetes

Also Published As

Publication number Publication date
WO2022132929A3 (fr) 2022-09-01

Similar Documents

Publication Publication Date Title
US20230272119A1 (en) Multispecific antibody molecules comprisinglambda and kappa light chains
JP7348899B2 (ja) 多重特異性分子及びその使用
AU2017238172B2 (en) Multispecific and multifunctional molecules and uses thereof
US20200071417A1 (en) Multispecific molecules and uses thereof
US20210246227A1 (en) Multispecific molecules that bind to myeloproliferative leukemia (mpl) protein and uses thereof
WO2019035938A1 (fr) Molécules multispécifiques se liant à bcma et leurs utilisations
AU2018260545B2 (en) Multispecific molecules comprising a non-immunoglobulin heterodimerization domain and uses thereof
WO2018151820A1 (fr) Molécules multifonctionnelles comprenant un ligand trimérique et leurs utilisations
WO2019178364A2 (fr) Molécules multifonctionnelles et utilisations associées
US20210238280A1 (en) Multifunctional molecules that bind to calreticulin and uses thereof
JP7465272B2 (ja) Ｃｓｆ１ｒ／ｃｃｒ２多特異性抗体
US20240002543A1 (en) Multifunctional molecules that bind to calreticulin and uses thereof
US20210380682A1 (en) Multifunctional molecules that bind to t cell related cancer cells and uses thereof
US20230102344A1 (en) Multifunctional molecules that bind to cd33 and uses thereof
WO2022132929A2 (fr) Molécules d'anticorps multispécifiques et leurs utilisations
JP2024084773A (ja) Ｃｓｆ１ｒ／ｃｃｒ２多特異性抗体

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21907714

Country of ref document: EP

Kind code of ref document: A2