WO2022159776A1 - Antibodies which bind human fibrin and methods of use - Google Patents

Antibodies which bind human fibrin and methods of use Download PDF

Info

Publication number
WO2022159776A1
WO2022159776A1 PCT/US2022/013423 US2022013423W WO2022159776A1 WO 2022159776 A1 WO2022159776 A1 WO 2022159776A1 US 2022013423 W US2022013423 W US 2022013423W WO 2022159776 A1 WO2022159776 A1 WO 2022159776A1
Authority
WO
WIPO (PCT)
Prior art keywords
seq
set forth
sequence set
cdr
antibody
Prior art date
Application number
PCT/US2022/013423
Other languages
French (fr)
Inventor
Jeffrey B. Stavenhagen
Olga GASIOROWSKA
Mathias Rickert
Aaron K. Sato
Pankaj Garg
Mary MATHIEU
Original Assignee
Therini Bio, 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 Therini Bio, Inc. filed Critical Therini Bio, Inc.
Publication of WO2022159776A1 publication Critical patent/WO2022159776A1/en

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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/55—Fab or Fab'
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • MS multiple sclerosis
  • perivascular microglia are believed to be necessary not only for the maintenance, but also for the onset of inflammatory demyelination in central nervous system (CNS) autoimmune disease.
  • CNS central nervous system
  • Activation of microglia contributes to both neuronal and oligodendrocyte death via release of cytokines and nitric oxide.
  • inflammatory processes are associated with destruction of myelin sheaths, and can also involve axonal damage that can lead to permanent functional deficits, such as paralysis and loss of vision.
  • Resident microglia are responsible for demyelination, via their ability to phagocytose myelin and secrete proinflammatory cytokines.
  • BBB disruption provokes the immediate and focal activation of microglia.
  • BBB disruption provokes the immediate and focal activation of microglia.
  • One of the earliest events coupled to BBB disruption in MS is leakage of the blood protein fibrinogen in the nervous system that results in perivascular deposition of fibrin. Fibrinogen is not present in the healthy CNS, but only leaks in the brain after BBB disruption, thus serving as an environmental "danger" signal.
  • CD1 lb/CD18 integrin receptor (also referred to as: Mac-1, aMfl 2, Complement Receptor 3) binds to the fibrin and induces microglial activation leading to inflammatory demyelination.
  • CD1 lb is the alpha chain of the receptor that regulates phagocytosis of myelin during inflammatory demyelination.
  • the y377-395 epitope of the fibrin or fibrinogen yC domain is the binding epitope of fibrin to CDIIb.
  • the fibrin y377-395 peptide functions as an inhibitor of microglia activation by blocking fibrin binding to Mac-1. Because fibrin mediates blood coagulation by binding via a distinct epitope to the platelet integrin allbp3 receptor, therapeutic agents (including antibodies), that block CD1 lb binding epitope to fibrin can reduce the damaging effects of fibrin in the nervous system without affecting its beneficial effects in blood coagulation. Therefore, safe, effective antibodies that inhibit fibrin induced microglial activation without affecting its beneficial effects in blood coagulation are needed as therapeutics for degenerative neuronal disorders that involve inflammatory demyelination.
  • isolated antibody that binds human fibrin, comprising a heavy chain comprising a variable heavy (VH) chain sequence comprising three heavy chain CDR sequences, CDR-H1, CDR-H2, and CDR-H3, and a light chain comprising a variable light (VL) chain sequence comprising three light chain CDR sequences, CDR-L1, CDR-L2, and CDR-L3, wherein: a.
  • CDR-H1 comprises the sequence set forth in SEQ ID NO: 1, 13, 25, 37, 49, 61, 73,
  • CDR-H2 comprises the sequence set forth in SEQ ID NO: 2, 14, 26, 38, 50, 62, 74,
  • CDR-H3 comprises the sequence set forth in SEQ ID NO: 3, 15, 27, 39, 51, 63, 75,
  • CDR-L1 comprises the sequence set forth in SEQ ID NO: 4, 16, 28, 40, 52, 64, 76,
  • CDR-L2 comprises the sequence set forth in SEQ ID NO: 5, 17, 29, 41, 53, 65, 77,
  • CDR-L3 comprises the sequence set forth in SEQ ID NO: 6, 18, 30, 42, 54, 66, 78,
  • the antibody comprises a VH sequence selected from a sequence set forth in one of SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562.
  • the antibody comprises a VL sequence selected from a sequence set forth in SEQ ID NO 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565.
  • the antibody comprises a VH sequence selected from a sequence set forth in one of SEQ ID Nos: 7, and the VL sequence set for in SEQ ID NO: 10.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 19 and a VL sequence set forth in SEQ ID NO: 22.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 31 and a VL sequence set forth in SEQ ID NO: 34.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 43 and a VL sequence set forth in SEQ ID NO: 46.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 55 and a VL sequence set forth in SEQ ID NO: 58. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 67 and a VL sequence set forth in SEQ ID NO: 70. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 67 and a VL sequence set forth in SEQ ID NO: 70. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 79 and a VL sequence set forth in SEQ ID NO: 82.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 79 and a VL sequence set forth in SEQ ID NO: 82. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 91 and a VL sequence set forth in SEQ ID NO: 94. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 103 and a VL sequence set forth in SEQ ID NO: 106. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 115 and a VL sequence set forth in SEQ ID NO: 118.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 127 and a VL sequence set forth in SEQ ID NO: 130. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 139 and a VL sequence set forth in SEQ ID NO: 142. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 151 and a VL sequence set forth in SEQ ID NO: 154. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 163 and a VL sequence set forth in SEQ ID NO: 166.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 175 and a VL sequence set forth in SEQ ID NO: 178. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 187 and a VL sequence set forth in SEQ ID NO: 190. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 199 and a VL sequence set forth in SEQ ID NO: 202. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 211 and a VL sequence set forth in SEQ ID NO: 214.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 223 and a VL sequence set forth in SEQ ID NO: 226. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 235 and a VL sequence set forth in SEQ ID NO: 238. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 250, and a VL sequence set forth in SEQ ID NO: 253. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 262 and a VL sequence set forth in SEQ ID NO: 265.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 274 and a VL sequence set forth in SEQ ID NO: 277. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 286 and a VL sequence set forth in SEQ ID NO: 289. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 298 and a VL sequence set forth in SEQ ID NO: 301. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 310, and a VL sequence set forth in SEQ ID NO: 313.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 322 and a VL sequence set forth in SEQ ID NO: 325. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 334 and a VL sequence set forth in SEQ ID NO: 337. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 346 and a VL sequence set forth in SEQ ID NO: 349. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 358 and a VL sequence set forth in SEQ ID NO: 361.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 370 and a VL sequence set forth in SEQ ID NO: 373. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 382 and a VL sequence set forth in SEQ ID NO: 385. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 394 and a VL sequence set forth in SEQ ID NO: 397. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 406 and a VL sequence set forth in SEQ ID NO: 409.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 418 and a VL sequence set forth in SEQ ID NO: 421. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 430 and a VL sequence set forth in SEQ ID NO: 433. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 442 and a VL sequence set forth in SEQ ID NO: 445. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 454 and a VL sequence set forth in SEQ ID NO: 457.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 466 and a VL sequence set forth in SEQ ID NO: 469. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 478 and a VL sequence set forth in SEQ ID NO: 481. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 490 and a VL sequence set forth in SEQ ID NO: 493. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 502 and a VL sequence set forth in SEQ ID NO: 505.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 514 and a VL sequence set forth in SEQ ID NO: 517. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 526 and a VL sequence set forth in SEQ ID NO: 529. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 538 and a VL sequence set forth in SEQ ID NO: 541. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 550 and a VL sequence set forth in SEQ ID NO: 553. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 562 and a VL sequence set forth in SEQ ID NO: 565.
  • the antibody is a humanized, human or chimeric antibody. In certain embodiments, the antibody is a humanized antibody. In certain embodiments, the antibody comprises a heavy chain human constant region of a class selected from IgG, IgA, IgD, IgE, and IgM. In certain embodiments, the human Fc region comprises a human heavy chain constant region of the class IgG and a subclass selected from IgGl, IgG2, IgG3, and IgG4. In certain embodiments, the human Fc region comprises wild-type, human IgGl Fc.
  • the human Fc domain comprises a sequence set forth in SEQ ID NO: 8, 20, 32, 44, 56, 68, 80, 92, 104, 116, 128, 140, 152, 164, 176, 188, 200, 212, 224, 236, 251, 263, 275, 287, 299, 311, 323, 335, 347, 359, 371, 383, 395, 407, 419, 431, 443, 455, 467, 479, 491, 503, 515, 527, 539, 551, or 563.
  • the heavy chain comprises a constant heavy chain sequence set forth by SEQ ID NO: 8, 20, 32, 44, 56, 68, 80, 92, 104, 116, 128, 140, 152, 164, 176, 188, 200, 212, 224, 236, 251, 263, 275, 287, 299, 311, 323, 335, 347, 359, 371, 383, 395, 407, 419, 431, 443, 455, 467, 479, 491, 503, 515, 527, 539, 551, or 563.
  • the light chain comprises a constant light chain sequence set forth by SEQ ID NO: 9, 21, 33, 45, 57, 69, 81, 93, 105, 117, 129, 141, 153, 165, 177, 189, 201, 213, 225, 237, 252, 264, 276, 288, 300, 312, 324, 336, 348, 360, 372, 384, 396, 408, 420, 432, 444, 456, 468, 480, 492, 504, 516, 528, 540, 552, or 564.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 7, and the VL sequence set forth in SEQ ID NO: 10; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 19, and the VL sequence set forth in SEQ ID NO: 22; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 31, and the VL sequence set forth in SEQ ID NO: 34; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 43, and the VL sequence set forth in SEQ ID NO: 46; and the human Fc region comprises wildtype, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 55, and the VL sequence set forth in SEQ ID NO: 58; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 67, and the VL sequence set forth in SEQ ID NO: 70; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 79, and the VL sequence set forth in SEQ ID NO: 82; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 91, and the VL sequence set forth in SEQ ID NO: 94; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 103, and the VL sequence set forth in SEQ ID NO: 106; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 115, and the VL sequence set forth in SEQ ID NO: 118; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 127, and the VL sequence set forth in SEQ ID NO: 130; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 139, and the VL sequence set forth in SEQ ID NO: 142; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 151, and the VL sequence set forth in SEQ ID NO: 154; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 163, and the VL sequence set forth in SEQ ID NO: 166; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 175, and the VL sequence set forth in SEQ ID NO: 178; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 187, and the VL sequence set forth in SEQ ID NO: 190; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 199, and the VL sequence set forth in SEQ ID NO: 202; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 211, and the VL sequence set forth in SEQ ID NO: 214; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 223, and the VL sequence set forth in SEQ ID NO: 226; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises the VH sequence set forth in SEQ ID NO: 235, and the VL sequence set forth in SEQ ID NO: 238; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 250, and a VL sequence set forth in SEQ ID NO: 253; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 262 and a VL sequence set forth in SEQ ID NO: 265; and the human Fc region comprises wildtype, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 274 and a VL sequence set forth in SEQ ID NO: 277; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 286 and a VL sequence set forth in SEQ ID NO: 289; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 298 and a VL sequence set forth in SEQ ID NO: 301; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 310, and a VL sequence set forth in SEQ ID NO: 313; and the human Fc region comprises wildtype, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 322 and a VL sequence set forth in SEQ ID NO: 325; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 334 and a VL sequence set forth in SEQ ID NO: 337; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 346 and a VL sequence set forth in SEQ ID NO: 349; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 358 and a VL sequence set forth in SEQ ID NO: 361; and the human Fc region comprises wildtype, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 370 and a VL sequence set forth in SEQ ID NO: 373; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 382 and a VL sequence set forth in SEQ ID NO: 385; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 394 and a VL sequence set forth in SEQ ID NO: 397; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 406 and a VL sequence set forth in SEQ ID NO: 409; and the human Fc region comprises wildtype, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 418 and a VL sequence set forth in SEQ ID NO: 421; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 430 and a VL sequence set forth in SEQ ID NO: 433; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 442 and a VL sequence set forth in SEQ ID NO: 445; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 454 and a VL sequence set forth in SEQ ID NO: 457; and the human Fc region comprises wildtype, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 466 and a VL sequence set forth in SEQ ID NO: 469; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 478 and a VL sequence set forth in SEQ ID NO: 481; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 490 and a VL sequence set forth in SEQ ID NO: 493; and the human Fc region comprises wild-type, human IgGl Fc.
  • wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 502 and a VL sequence set forth in SEQ ID NO: 505; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 514 and a VL sequence set forth in SEQ ID NO: 517; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 526 and a VL sequence set forth in SEQ ID NO: 529; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 538 and a VL sequence set forth in SEQ ID NO: 541; and the human Fc region comprises wildtype, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 550 and a VL sequence set forth in SEQ ID NO: 553; and the human Fc region comprises wild-type, human IgGl Fc.
  • the antibody comprises a VH sequence set forth in SEQ ID NO: 562 and a VL sequence set forth in SEQ ID NO: 565; and the human Fc region comprises wild-type, human IgGl Fc.
  • the Fc region comprises one or more amino acid substitutions, wherein the one or more substitutions result in increased antibody half-life, increased ADCC activity, increased ADCP activity, or increased CDC activity compared with the Fc without the one or more substitutions, the Fc region binds an Fey Receptor selected from the group consisting of: FcyRI, FcyRIIa, FcyRIIb, FcyRIIc, FcyRIIIa, and FcyRIIIb. the Fc region binds an Fey Receptor selected from the group consisting of: FcyRI, FcyRIIa, FcyRIIb, FcyRIIc, FcyRIIIa, and FcyRIIIb. In certain embodiments, the Fc region has reduced or is lacking Fc effector function compared to a wild-type Fc region.
  • the antibody binds Fibrin with a KD of less than or equal to about 1 xlO' 6 M, as measured by enzyme-linked immunosorbent assay (ELISA).
  • ELISA enzyme-linked immunosorbent assay
  • the antibody is a monoclonal antibody.
  • the antibody binds Fibrin Gamma Globular (FGG) domain. In certain embodiments, the antibody binds FGG with a KD of less than or equal to about 1 xl0-8 M, as measured by ELISA.
  • FGG Fibrin Gamma Globular
  • the antibody binds to a peptide comprising SEQ ID NO: 241 with a KD of less than or equal to about 1 xl0-8 M, as measured by ELISA.
  • the antibody binds to fibrinogen with a KD of greater than or equal to about 1 xl0-8 M, as measured by ELISA. In certain embodiments, the antibody inhibits Mac-1 binding to fibrin or fibrinogen yC domain. In certain embodiments, the antibody exhibits inhibition of microglial adhesion to the fibrin or fibrinogen yC domain.
  • the antibody exhibits activation of CD1 IB positive monocytic cells.
  • the antibodies described herein are used for the treatment of a degenerative disorder of the nervous system.
  • polynucleotide or set of polynucleotides encoding the antibody of any of the above claims, a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen-binding portion thereof, and optionally, wherein the polynucleotide or set of polynucleotides comprises cDNA.
  • polynucleotide or set of polynucleotides encoding the antibody of any of the above claims, a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen-binding portion thereof, and optionally, wherein the polynucleotide or set of polynucleotides comprises cDNA.
  • a vector or set of vectors comprising the polynucleotide or set of polynucleotides encoding the antibody of any of the above claims, a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen-binding portion thereof, and optionally, wherein the polynucleotide or set of polynucleotides comprises cDNA.
  • a host cell comprising the polynucleotide or set of polynucleotides, or a vector or set of vectors, encoding the antibody of any of the above claims, a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen- binding portion thereof, and optionally, wherein the polynucleotide or set of polynucleotides comprises cDNA.
  • a method of producing an antibody the method comprising expressing the antibody with the host cell and isolating the expressed antibody.
  • described herein is a pharmaceutical composition
  • a pharmaceutical composition comprising an antibody described herein and a pharmaceutically acceptable excipient.
  • kits comprising the antibody described herein and instructions for use.
  • described herein is a method for treating a degenerative disorder of the nervous system, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein.
  • the degenerative disorder of the nervous system is selected from the group consisting of: multiple sclerosis, spinal cord injury, stroke, and Alzheimer’s Disease.
  • described herein is a method of treating a pathology associated with Mac-1 binding to fibrin or Mac-1 binding with fibrinogen, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein.
  • described herein is a method of inhibiting microglia activation, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein.
  • described herein is a method of preventing a degenerative disorder of the nervous system, the method comprising administering to a mammalian subject a therapeutically effective amount of the antibody or pharmaceutical composition described herein.
  • described herein is a method of treating colitis in a subject in need thereof, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein.
  • described herein is a method of preventing colitis in a subject in need thereof, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein.
  • Figure l is a diagram illustrating the protocol used for screening antibodies using two phage libraries and three panning arms with five rounds of selection.
  • Figure 2 depicts a table and graphs showing results of phage display screen.
  • the table (top) shows increase in binding to FGG (Fibrinogen Gamma Globular domain) after each round of selection of an exemplary clone.
  • the three graphs depict binding of 33 exemplary clones by ELISA to FGG, Fibrinogen, and the ratio of FGG/Fibrinogen binding (bottom).
  • Figure 3 is a table depicting the results of the phage display screens from either the hyperimmune Fab phage library or the structural scFv phage library, showing the number of unique clones obtained wih a ratio of FGG/Fibrinogen binding greater than 2.
  • Figure 4 are graphs depicting binding of selected clones to P2 peptide (top-left), FGG (top-right), Fibrin (bottom-left), and Fibrinogen (bottom-right).
  • Figure 5 are graphs depicting binding of selected clones to Fibrin (top), and FGG (bottom).
  • Figure 6 are graphs depicting binding of selected clones to Fibrin (top-left), FGG (top-right), and P2 peptide (bottom).
  • Figure 7 are graphs depicting binding of selected clones to Fibrin (top), and FGG (bottom).
  • Figure 8 are graphs depicting binding of selected clones to Fibrin (top-left), FGG (top-right), and P2 peptide (bottom).
  • Figure 9 are graphs depicting binding of selected clones to Fibrin (top-left), FGG (top-right), and P2 peptide (bottom).
  • Figure 10 are graphs depicting binding of selected clones to Fibrin (top), and FGG (bottom).
  • Figure 11 is a graph depicting binding of select clones to recombinant fibrinogen gamma chain (biotinylated- histidine tagged-FGG-F168A/F303A).
  • Figure 12 is a graph depicting reduction of clinical score of a rodent experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis in mice treated with 15 mg per kg of the indicated antibody clone.
  • EAE experimental autoimmune encephalomyelitis
  • compositions described herein include “comprising,” “consisting,” and “consisting essentially of’ aspects and embodiments. [0047] For all compositions described herein, and all methods using a composition described herein, the compositions can either comprise the listed components or steps, or can “consist essentially of’ the listed components or steps.
  • composition when a composition is described as “consisting essentially of’ the listed components, the composition contains the components listed, and may contain other components which do not substantially affect the condition being treated, but do not contain any other components which substantially affect the condition being treated other than those components expressly listed; or, if the composition does contain extra components other than those listed which substantially affect the condition being treated, the composition does not contain a sufficient concentration or amount of the extra components to substantially affect the condition being treated.
  • a method is described as “consisting essentially of’ the listed steps, the method contains the steps listed, and may contain other steps that do not substantially affect the condition being treated, but the method does not contain any other steps which substantially affect the condition being treated other than those steps expressly listed.
  • composition when a composition is described as ‘consisting essentially of a component, the composition may additionally contain any amount of pharmaceutically acceptable carriers, vehicles, or diluents and other such components which do not substantially affect the condition being treated.
  • vector refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked.
  • the term includes the vector as a selfreplicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced.
  • Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as “expression vectors.”
  • host cell refers to cells into which an exogenous nucleic acid has been introduced, and the progeny of such cells.
  • Host cells include “transformants” (or “transformed cells”) and “transfectants” (or “transfected cells”), which each include the primary transformed or transfected cell and progeny derived therefrom.
  • Such progeny may not be completely identical in nucleic acid content to a parent cell, and may contain mutations.
  • a “recombinant host cell” or “host cell” refers to a cell that includes an exogenous polynucleotide, regardless of the method used for insertion, for example, direct uptake, transduction, f-mating, or other methods known in the art to create recombinant host cells.
  • the term “eukaryote” refers to organisms belonging to the phylogenetic domain Eucarya such as animals (including but not limited to, mammals, insects, reptiles, birds, etc.), ciliates, plants (including but not limited to, monocots, dicots, algae, etc.), fungi, yeasts, flagellates, microsporidia, protists, etc.
  • prokaryote refers to prokaryotic organisms.
  • a non-eukaryotic organism can belong to the Eubacteria (including but not limited to, Escherichia coli, Thermus thermophilus, Bacillus stearothermophilus, Pseudomonas fluorescens, Pseudomonas aeruginosa, Pseudomonas putida, etc.) phylogenetic domain, or the Archaea (including but not limited to, Methanococcus jannaschii, Methanobacterium thermoautotrophicum, Halobacterium such as Haloferax volcanii and Halobacterium species NRC-1, Archaeoglobus fulgidus, Pyrococcus furiosus, Pyrococcus horikoshii, Aeuropyrum pernix, etc.) phylogenetic domain.
  • Eubacteria including but not limited to, Escherichia coli, Thermus thermophilus,
  • an “effective amount” or “therapeutically effective amount” as used herein refers to an amount of therapeutic compound, such as an anti-FIBRIN antibody, administered to an individual, either as a single dose or as part of a series of doses, which is effective to produce or contribute to a desired therapeutic effect, either alone or in combination with another therapeutic modality. Examples of a desired therapeutic effect is enhancing an immune response, slowing or delaying tumor development; stabilization of disease; amelioration of one or more symptoms. An effective amount may be given in one or more dosages.
  • treating refers to clinical intervention in an attempt to alter the natural course of a disease or condition in a subject in need thereof. Treatment can be performed during the course of clinical pathology. Desirable effects of treatment include preventing recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.
  • sufficient amount means an amount sufficient to produce a desired effect, e.g., an amount sufficient to modulate an immune response in a subject.
  • the term “subject” or “individual” means a mammalian subject. Exemplary subjects include humans, monkeys, dogs, cats, mice, rats, cows, horses, camels, goats, rabbits, and sheep. In certain embodiments, the subject is a human. In some embodiments the subject has a disease or condition that can be treated with an antibody provided herein. In some aspects, the disease or condition is a cancer. In some aspects, the disease or condition is a viral infection.
  • in vitro refers to processes that occur in a living cell growing separate from a living organism, e.g., growing in tissue culture.
  • in vivo refers to processes that occur in a living organism.
  • kits therapeutic or diagnostic products
  • pharmaceutical composition refers to a preparation which is in such form as to permit the biological activity of an active ingredient contained therein to be effective in treating a subject, and which contains no additional components which are unacceptably toxic to the subject in the amounts provided in the pharmaceutical composition.
  • co-administration include the administration of two or more therapeutic agents either simultaneously, concurrently or sequentially within no specific time limits.
  • the agents are present in the cell or in the subject's body at the same time or exert their biological or therapeutic effect at the same time.
  • the therapeutic agents are in the same composition or unit dosage form. In other embodiments, the therapeutic agents are in separate compositions or unit dosage forms.
  • a first agent can be administered prior to the administration of a second therapeutic agent.
  • modulate and “modulation” refer to reducing or inhibiting or, alternatively, activating or increasing, a recited variable.
  • increase and activate refer to an increase of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or greater in a recited variable.
  • reduce and “inhibit” refer to a decrease of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50- fold, 100-fold, or greater in a recited variable.
  • the term “about” indicates and encompasses an indicated value and a range above and below that value. In certain embodiments, the term “about” indicates the designated value ⁇ 10%, ⁇ 5%, or ⁇ 1%. In certain embodiments, where applicable, the term “about” indicates the designated value(s) ⁇ one standard deviation of that value(s).
  • agonist refers to the activation of receptor signaling to induce a biological response associated with activation of the receptor.
  • agonist is an entity that binds to and agonizes a receptor.
  • antiagonize refers to the inhibition of receptor signaling to inhibit a biological response associated with activation of the receptor.
  • An “antagonist” is an entity that binds to and antagonizes a receptor.
  • amino acid refers to the twenty common naturally occurring amino acids.
  • Naturally occurring amino acids include alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine (Cys; C); glutamic acid (Glu; E), glutamine (Gin; Q), Glycine (Gly; G); histidine (His; H), isoleucine (He; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Vai; V).
  • affinity refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen or epitope). Unless indicated otherwise, as used herein, “affinity” refers to intrinsic binding affinity, which reflects a 1 : 1 interaction between members of a binding pair (e.g., antibody and antigen or epitope).
  • kd (sec-1), as used herein, refers to the dissociation rate constant of a particular antibody - antigen interaction. This value is also referred to as the koff value.
  • ka (M-l *sec-l), as used herein, refers to the association rate constant of a particular antibody -antigen interaction. This value is also referred to as the kon value.
  • KD kd/ka.
  • affinity of an antibody is described in terms of the KD for an interaction between such antibody and its antigen. For clarity, as known in the art, a smaller KD value indicates a higher affinity interaction, while a larger KD value indicates a lower affinity interaction.
  • antibody is used herein in its broadest sense and includes certain types of immunoglobulin molecules comprising one or more antigen-binding domains that specifically bind to an antigen or epitope.
  • An antibody specifically includes intact antibodies (e.g., intact immunoglobulins), antibody fragments, and multi-specific antibodies.
  • a “Fibrin antibody,” “anti-Fibrin antibody,” or “Fibrin -specific antibody” is an antibody, as provided herein, which specifically binds to the antigen Fibrin. In some embodiments, the antibody binds the extracellular domain of Fibrin. In certain embodiments, a Fibrin antibody provided herein binds to an epitope of Fibrin that is conserved between or among Fibrin proteins from different species.
  • epitope means a portion of an antigen that specifically binds to an antibody.
  • hypervariable region refers to each of the regions of an antibody variable domain which are hypervariable in sequence and/or form structurally defined loops (“hypervariable loops”).
  • antigen-binding domain means the portion of an antibody that is capable of specifically binding to an antigen or epitope.
  • chimeric antibody refers to an antibody in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.
  • human antibody refers to an antibody which possesses an amino acid sequence corresponding to that of an antibody produced by a human or a human cell, or derived from a non-human source that utilizes a human antibody repertoire or human antibody-encoding sequences (e.g., obtained from human sources or designed de novo). Human antibodies specifically exclude humanized antibodies.
  • humanized antibody refers to a protein having a sequence that differs from the sequence of an antibody derived from a non-human species by one or more amino acid substitutions, deletions, and/or additions, such that the humanized antibody is less likely to induce an immune response, and/or induces a less severe immune response, as compared to the non-human species antibody, when it is administered to a human subject.
  • multispecific antibody refers to an antibody that comprises two or more different antigen-binding domains that collectively specifically bind two or more different epitopes.
  • a “monospecific antibody” is an antibody that comprises one or more binding sites that specifically bind to a single epitope.
  • An example of a monospecific antibody is a naturally occurring IgG molecule which, while divalent (i.e., having two antigen-binding domains), recognizes the same epitope at each of the two antigen-binding domains.
  • the binding specificity may be present in any suitable valency.
  • the term “monoclonal antibody” refers to an antibody from a population of substantially homogeneous antibodies.
  • a population of substantially homogeneous antibodies comprises antibodies that are substantially similar and that bind the same epitope(s), except for variants that may normally arise during production of the monoclonal antibody. Such variants are generally present in only minor amounts.
  • a monoclonal antibody is typically obtained by a process that includes the selection of a single antibody from a plurality of antibodies.
  • the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, yeast clones, bacterial clones, or other recombinant DNA clones.
  • the selected antibody can be further altered, for example, to improve affinity for the target (“affinity maturation”), to humanize the antibody, to improve its production in cell culture, and/or to reduce its immunogenicity in a subject.
  • the term "single-chain” refers to a molecule comprising amino acid monomers linearly linked by peptide bonds.
  • the C-terminus of the Fab light chain is connected to the N-terminus of the Fab heavy chain in the single-chain Fab molecule.
  • an scFv has a variable domain of light chain (VL) connected from its C-terminus to the N-terminal end of a variable domain of heavy chain (VH) by a polypeptide chain.
  • VL variable domain of light chain
  • VH variable domain of heavy chain
  • the scFv comprises of polypeptide chain where in the C-terminal end of the VH is connected to the N-terminal end of VL by a polypeptide chain.
  • the “Fab fragment” (also referred to as fragment antigen-binding) contains the constant domain (CL) of the light chain and the first constant domain (CHI) of the heavy chain along with the variable domains VL and VH on the light and heavy chains respectively.
  • the variable domains comprise the complementarity determining loops (CDR, also referred to as hypervariable region) that are involved in antigen-binding.
  • CDR complementarity determining loops
  • Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CHI domain including one or more cysteines from the antibody hinge region.
  • F(ab’)2 fragments contain two Fab’ fragments joined, near the hinge region, by disulfide bonds.
  • F(ab’)2 fragments may be generated, for example, by recombinant methods or by pepsin digestion of an intact antibody.
  • the F(ab’) fragments can be dissociated, for example, by treatment with B-mercaptoethanol.
  • Fv fragments comprise a non-covalently-linked dimer of one heavy chain variable domain and one light chain variable domain.
  • Single-chain Fv or “sFv” or “scFv” includes the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain.
  • the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen-binding.
  • HER2 antibody scFv fragments are described in WO93/16185; U.S. Pat. No. 5,571,894; and U.S. Pat. No. 5,587,458.
  • scFv-Fc fragments comprise an scFv attached to an Fc domain.
  • an Fc domain may be attached to the C-terminal of the scFv.
  • the Fc domain may follow the VH or VL, depending on the orientation of the variable domains in the scFv (i.e., VH -VL or VL - VH ). Any suitable Fc domain known in the art or described herein may be used.
  • the Fc domain comprises an IgG4 Fc domain.
  • single domain antibody refers to a molecule in which one variable domain of an antibody specifically binds to an antigen without the presence of the other variable domain.
  • Single domain antibodies, and fragments thereof, are described in Arabi Ghahroudi et al., FEBS Letters, 1998, 414:521-526 and Muyldermans et al., Trends in Biochem. Sci., 2001, 26:230-245, each of which is incorporated by reference in its entirety.
  • Single domain antibodies are also known as sdAbs or nanobodies. Sdabs are fairly stable and easy to express as fusion partner with the Fc chain of an antibody (Harmsen MM, De Haard HJ (2007). "Properties, production, and applications of camelid single-domain antibody fragments". Appl. Microbiol Biotechnol. 77(1): 13-22).
  • full length antibody is an antibody having a structure substantially similar to a naturally occurring antibody structure and having heavy chains that comprise an Fc region.
  • a “full length antibody” is an antibody that comprises two heavy chains and two light chains.
  • antibody fragment refers to an antibody that comprises a portion of an intact antibody, such as the antigen-binding or variable region of an intact antibody.
  • Antibody fragments include, for example, Fv fragments, Fab fragments, F(ab’)2 fragments, Fab’ fragments, scFv (sFv) fragments, and scFv-Fc fragments.
  • Fc domain or “Fc region” herein is used to define a C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region.
  • the term includes native sequence Fc regions and variant Fc regions.
  • substantially purified refers to a construct described herein, or variant thereof that may be substantially or essentially free of components that normally accompany or interact with the protein as found in its naturally occurring environment, i.e. a native cell, or host cell in the case of recombinantly produced heteromultimer that in certain embodiments, is substantially free of cellular material includes preparations of protein having less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1% (by dry weight) of contaminating protein.
  • percent “identity,” in the context of two or more nucleic acid or polypeptide sequences, refer to two or more sequences or subsequences that have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., using publicly available computer software such as BLAST, BLASTP, BLASTN, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, CLUSTAL OMEGA, or MUSCLE software or other algorithms available to persons of skill) or by visual inspection.
  • sequence comparison algorithms e.g., using publicly available computer software such as BLAST, BLASTP, BLASTN, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, CLUSTAL OMEGA, or MUSCLE software or other algorithms available to persons of skill
  • BLAST analyses Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (ncbi.nlm.nih.gov). Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. Depending on the application, the percent "identity" can exist over a region of the sequence being compared, e.g., over a functional domain, or, alternatively, exist over the full length of the two sequences to be compared.
  • sequence comparison typically one sequence acts as a reference sequence to which test sequences are compared.
  • test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated.
  • sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters.
  • Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see generally Ausubel et al., infra).
  • Ranges recited herein are understood to be shorthand for all of the values within the range, inclusive of the recited endpoints.
  • a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50.
  • the present application provides antibodies and compositions comprising an antibody which binds a fibrin protein.
  • the recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as the myriad immunoglobulin variable region genes.
  • Light chains are classified as either kappa or lambda.
  • the "class" of an antibody or immunoglobulin refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2.
  • the heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, 6, a, y, and p, respectively.
  • An exemplary immunoglobulin (antibody) structural unit is composed of two pairs of polypeptide chains, each pair having one “light” (about 25 kD) and one “heavy” chain (about 50-70 kD).
  • the N-terminal domain of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition.
  • the terms variable light chain (VL) and variable heavy chain (VH) refer to these light and heavy chain domains respectively.
  • the IgGl heavy chain comprises of the VH, CHI, CH2 and CH3 domains respectively from the N to C-terminus.
  • the light chain comprises of the VL and CL domains from N to C terminus.
  • the IgGl heavy chain comprises a hinge between the CHI and CH2 domains.
  • the immunoglobulin constructs comprise at least one immunoglobulin domain from IgG, IgM, IgA, IgD, or IgE connected to a therapeutic polypeptide.
  • the immunoglobulin domain found in an antibody provided herein is from or derived from an immunoglobulin based construct such as a diabody, or a nanobody.
  • the immunoglobulin constructs described herein comprise at least one immunoglobulin domain from a heavy chain antibody such as a camelid antibody.
  • the immunoglobulin constructs provided herein comprise at least one immunoglobulin domain from a mammalian antibody such as a bovine antibody, a human antibody, a camelid antibody, a mouse antibody or any chimeric antibody.
  • the antibodies provided herein comprise a heavy chain.
  • the heavy chain is an IgA.
  • the heavy chain is an IgD.
  • the heavy chain is an IgE.
  • the heavy chain is an IgG.
  • the heavy chain is an IgM.
  • the heavy chain is an IgGl.
  • the heavy chain is an IgG2.
  • the heavy chain is an IgG3.
  • the heavy chain is an IgG4.
  • the heavy chain is an IgAl. In one embodiment, the heavy chain is an IgA2.
  • an antibody is an IgGl antibody. In some embodiments, an antibody is an IgG3 antibody. In some embodiments, an antibody is an IgG2 antibody. In some embodiments, an antibody is an IgG4 antibody.
  • native four-chain antibodies comprise six HVRs; three in the VH (Hl, H2, H3), and three in the VL (LI, L2, L3).
  • HVRs generally comprise amino acid residues from the hypervariable loops and/or from the complementarity determining regions (CDRs), the latter being of highest sequence variability and/or involved in antigen recognition. With the exception of CDR1 in VH, CDRs generally comprise the amino acid residues that form the hypervariable loops.
  • Hypervariable regions (HVRs) are also referred to as “complementarity determining regions” (CDRs), and these terms are used herein interchangeably in reference to portions of the variable region that form the antigen-binding regions.
  • amino acid sequence boundaries of a CDR can be determined by one of skill in the art using any of a number of known numbering schemes, including those described by Kabat et al., supra (“Kabat” numbering scheme); Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 (“Chothia” numbering scheme); MacCallum et al., 1996, J. Mol. Biol. 262:732- 745 (“Contact” numbering scheme); Lefranc et al., Dev. Comp. Immunol., 2003, 27:55-77 (“IMGT” numbering scheme); and Honegge and Pliickthun, J. Mol. Biol., 2001, 309:657-70 (“AHo” numbering scheme); each of which is incorporated by reference in its entirety.
  • Table A provides the positions of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 as identified by the Kabat and Chothia schemes.
  • residue numbering is provided using both the Kabat and Chothia numbering schemes.
  • CDRs may be assigned, for example, using antibody numbering software, such as Abnum, available at www.bioinf.org.uk/abs/abnum/, and described in Abhinandan and Martin, Immunology, 2008, 45:3832-3839, incorporated by reference in its entirety.
  • EU numbering scheme is generally used when referring to a residue in an antibody heavy chain constant region (e.g., as reported in Kabat et al., supra). Unless stated otherwise, the EU numbering scheme is used to refer to residues in antibody heavy chain constant regions described herein.
  • an antigen-binding domain is an antigen-binding domain formed by a VH-VL dimer of an antibody.
  • Another example of an antigen-binding domain is an antigen-binding domain formed by diversification of certain loops from the tenth fibronectin type III domain of an Adnectin.
  • An antigen-binding domain can include CDRs 1, 2, and 3 from a heavy chain in that order; and CDRs 1, 2, and 3 from a light chain in that order.
  • Epitopes frequently consist of surface-accessible amino acid residues and/or sugar side chains and may have specific three-dimensional structural characteristics, as well as specific charge characteristics. Conformational and non-conformational epitopes are distinguished in that the binding to the former but not the latter may be lost in the presence of denaturing solvents.
  • An epitope may comprise amino acid residues that are directly involved in the binding, and other amino acid residues, which are not directly involved in the binding.
  • the epitope to which an antibody binds can be determined using known techniques for epitope determination such as, for example, testing for antibody binding to Fibrin variants with different point-mutations, or to chimeric Fibrin variants.
  • an epitope mapping can be performed by methods known in the art.
  • Chimeric antibodies are antibodies in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.
  • Human antibodies are antibodies which possesses an amino acid sequence corresponding to that of an antibody produced by a human or a human cell, or derived from a non-human source that utilizes a human antibody repertoire or human antibody-encoding sequences (e.g., obtained from human sources or designed de novo). Human antibodies specifically exclude humanized antibodies.
  • a humanized antibody has a sequence that differs from the sequence of an antibody derived from a non-human species by one or more amino acid substitutions, deletions, and/or additions, such that the humanized antibody is less likely to induce an immune response, and/or induces a less severe immune response, as compared to the non-human species antibody, when it is administered to a human subject.
  • certain amino acids in the framework and constant domains of the heavy and/or light chains of the non- human species antibody are mutated to produce the humanized antibody.
  • the constant domain(s) from a human antibody are fused to the variable domain(s) of a non-human species.
  • one or more amino acid residues in one or more CDR sequences of a non-human antibody are changed to reduce the likely immunogenicity of the non-human antibody when it is administered to a human subject, wherein the changed amino acid residues either are not critical for immunospecific binding of the antibody to its antigen, or the changes to the amino acid sequence that are made are conservative changes, such that the binding of the humanized antibody to the antigen is not significantly worse than the binding of the non-human antibody to the antigen. Examples of how to make humanized antibodies can be found in U.S. Pat. Nos. 6,054,297, 5,886,152 and 5,877,293.
  • the two or more different epitopes may be epitopes on the same antigen (e.g., a single Fibrin molecule expressed by a cell) or on different antigens (e.g., different Fibrin molecules expressed by the same cell, or a Fibrin molecule and a non- Fibrin molecule).
  • a multi-specific antibody binds two different epitopes (i.e., a “bispecific antibody”).
  • a multi-specific antibody binds three different epitopes (i.e., a “trispecific antibody”).
  • Anti-Fibrin antibodies can include those described herein such as the clones set forth in the drawings and/or tables.
  • the antibody comprises an alternative scaffold.
  • the antibody consists of an alternative scaffold.
  • the antibody consists essentially of an alternative scaffold.
  • the antibody comprises an antibody fragment.
  • the antibody consists of an antibody fragment.
  • the antibody consists essentially of an antibody fragment.
  • the antibodies are monoclonal antibodies.
  • the antibodies are polyclonal antibodies.
  • the antibodies are produced by hybridomas. In other embodiments, the antibodies are produced by recombinant cells engineered to express the desired variable and constant domains.
  • the antibodies may be single chain antibodies or other antibody derivatives retaining the antigen specificity and the lower hinge region or a variant thereof.
  • the antibodies may be polyfunctional antibodies, recombinant antibodies, human antibodies, humanized antibodies, fragments or variants thereof.
  • the antibody fragment or a derivative thereof is selected from a Fab fragment, a Fab '2 fragment, a CDR and ScFv.
  • the antibodies are capable of forming an immune complex.
  • an immune complex can be a tumor cell covered by antibodies.
  • sequence comparison typically one sequence acts as a reference sequence to which test sequences are compared.
  • test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated.
  • sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters.
  • Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nafl. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see generally Ausubel et al., infra).
  • an antibody provided herein comprises a VH sequence selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562.
  • an antibody provided herein comprises a VH sequence having at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% identity to an illustrative VH sequence provided in SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562.
  • an antibody provided herein comprises a VH sequence provided in SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562, with up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acid substitutions.
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises a VL sequence selected from SEQ ID NOs: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397,
  • an antibody provided herein comprises a VL sequence having at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% identity to an illustrative VL sequence provided in SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373,
  • an antibody provided herein comprises a VL sequence provided in SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226,
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises a VH sequence selected from SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562; and a VL sequence selected from SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397,
  • 562 can be combined with any of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565.
  • an antibody provided herein comprises a VH sequence having at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% identity to an illustrative VH sequence provided in SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382,
  • VL sequence having at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% identity to an illustrative VL sequence provided in SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118,
  • an antibody provided herein comprises a VH sequence provided in SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562, with up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acid substitutions, and a VL sequence provided in SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises one to three CDRs of a VH domain selected from SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562.
  • an antibody provided herein comprises two to three CDRs of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562.
  • an antibody provided herein comprises three CDRs of a VH domain selected from SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562.
  • the CDRs are Exemplary CDRs.
  • the CDRs are Kabat CDRs.
  • the CDRs are Chothia CDRs.
  • the CDRs are AbM CDRs.
  • the CDRs are Contact CDRs.
  • the CDRs are IMGT CDRs.
  • the CDRs are CDRs having at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1, CDR-H2, or CDR-H3 of SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490,
  • the CDR-H1 is a CDR-H1 of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370,
  • the CDR-H2 is a CDR-H2 of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions.
  • the CDR-H3 is a CDR-H3 of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions.
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises one to three CDRs of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385,
  • an antibody provided herein comprises two to three CDRs of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433,
  • an antibody provided herein comprises three CDRs of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565.
  • the CDRs are Exemplary CDRs. In some aspects, the CDRs are Kabat CDRs. In some aspects, the CDRs are Chothia CDRs. In some aspects, the CDRs are AbM CDRs. In some aspects, the CDRs are Contact CDRs. In some aspects, the CDRs are IMGT CDRs.
  • the CDRs are CDRs having at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1, CDR-L2, or CDR-L3 of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565.
  • the CDR-L1 is a CDR-L1 of a VL domain of SEQ ID NO: 21, with up to 1, 2, 3, 4, or 5 amino acid substitutions.
  • the CDR-L2 is a CDR-L2 of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions.
  • the CDR-L3 is a CDR-L3 of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions.
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises one to three CDRs of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358,
  • an antibody provided herein comprises two to three CDRs of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358,
  • an antibody provided herein comprises three CDRs of a VH domain selected from SEQ ID NOs: , 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382,
  • the CDRs are Exemplary CDRs. In some aspects, the CDRs are Kabat CDRs. In some aspects, the CDRs are Chothia CDRs. In some aspects, the CDRs are AbM CDRs. In some aspects, the CDRs are Contact CDRs. In some aspects, the CDRs are IMGT CDRs.
  • an antibody provided herein comprises a CDR-H3 selected of SEQ ID NOs: 3, 15, 27, 39, 51, 63, 75, 87, 99, 111, 123, 135, 147, 159, 171, 183, 195, 207, 219, or 231, 246, 258, 270, 282, 294, 306, 318, 330, 342, 354, 366, 379, 390, 402, 414, 426, 438, 450, 462, 474, 486, 498, 510, 522, 534, 548, or 558.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID
  • the CDR-H3 is a
  • CDR-H3 selected of SEQ ID NO:3, 15, 27, 39, 51, 63, 75, 87, 99, 111, 123, 135, 147, 159, 171, 183, 195, 207, 219, or 231, 246, 258, 270, 282, 294, 306, 318, 330, 342, 354, 366, 379, 390, 402, 414, 426, 438, 450, 462, 474, 486, 498, 510, 522, 534, 548, or 558, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions.
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises a CDR-H1 of SEQ ID NO: 1, 13, 25, 37, 49, 61, 73, 85, 97, 109, 121, 133, 145, 157, 169, 181, 193, 205, 217, 229, 244, 256, 268, 280, 292, 304, 316, 328, 340, 352, 364, 376, 388, 400, 412, 424, 436,
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 1, 13, 25, 37, 49, 61, 73, 85, 97, 109, 121, 133, 145, 157, 169, 181, 193, 205, 217, 229, 244,
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 1, 13, 25, 37, 49, 61, 73, 85, 97, 109, 121, 133, 145, 157, 169, 181, 193, 205, 217, 229, 244, 256, 268, 280, 292, 304, 316, 328, 340, 352, 364, 376, 388, 400, 412, 424, 436, 448, 460, 472, 484, 496, 508, 520, 532, 546, or 556, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions.
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises a CDR-H2 selected of SEQ ID NO: 2, 14, 26, 38, 50, 62, 74, 86, 98, 110, 122, 134, 146, 158, 170, 182, 194, 206, 218, 230, 245, 257, 269, 281, 293, 305, 317, 329, 341, 353, 365, 378, 389, 401, 413, 425,
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 2, 14, 26, 38, 50, 62, 74, 86, 98, 110, 122, 134, 146, 158, 170, 182, 194, 206, 218, 230, 245,
  • the CDR-H2 is a CDR-H2 selected of SEQ ID NO: 2, 14, 26, 38, 50, 62, 74, 86, 98, 110, 122, 134, 146, 158, 170, 182, 194, 206, 218, 230, 245, 257, 269, 281, 293, 305, 317, 329, 341, 353, 365, 378, 389, 401, 413, 425, 437, 449, 461, 473, 485, 497, 509, 521, 533, 547, or 557, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions.
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises a CDR-L3 of SEQ ID NO: 6, 18, 30, 42, 54, 66, 78, 90, 102, 114, 126, 138, 150, 162, 174, 186, 198, 210, 222, 234, 249, 261, 273, 285, 297, 309, 321, 333, 345, 357, 369, 382, 393, 405, 417, 429, 441, 453,
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 6, 18, 30, 42, 54, 66, 78, 90, 102, 114, 126, 138, 150, 162, 174, 186, 198, 210, 222, 234, 249, 261, 273, 285, 297, 309, 321, 333, 345, 357, 369, 382, 393, 405, 417, 429, 441, 453, 465, 477, 489,
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 6, 18, 30, 42, 54, 66, 78, 90, 102, 114, 126, 138, 150, 162, 174, 186, 198, 210, 222, 234, 249, 261, 273, 285, 297, 309, 321, 333, 345, 357, 369, 382, 393, 405, 417, 429, 441, 453, 465, 477, 489, 501, 513, 525, 537, 551, or 561, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions.
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises a CDR-L2 of SEQ ID NO: 5, 17, 29, 41, 53, 65, 77, 89, 101, 113, 125, 137, 149, 161, 173, 185, 197, 209, 221, 233, 248, 260, 272, 284, 296, 308, 320, 332, 344, 356, 368, 381, 392, 404, 416, 428, 440, 452, 464, 476, 488, 500, 512, 524, 536, 550, or 560.
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 5, 17, 29, 41, 53, 65, 77, 89, 101, 113, 125, 137, 149, 161, 173, 185, 197, 209, 221, 233, 248, 260, 272, 284, 296, 308, 320, 332, 344, 356, 368, 381, 392, 404, 416, 428, 440, 452, 464, 476, 488,
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 1
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises a CDR-L1 of SEQ ID NO: 4, 16, 28, 40, 52, 64, 76, 88, 100, 112, 124, 136, 148, 160, 172, 184, 196, 208, 220, 232, 247, 259, 271, 283, 295, 307, 319, 331, 343, 355, 367, 380, 391, 403, 415, 427, 439, 451, 463, 475, 487, 499, 511, 523, 535, 549, or 559.
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 4, 16, 28, 40, 52, 64, 76, 88, 100, 112, 124, 136, 148, 160, 172, 184, 196, 208, 220, 232, 247, 259, 271, 283, 295, 307, 319, 331, 343, 355, 367, 380, 391, 403, 415, 427, 439, 451, 463, 475, 487, 499, 511, 523, 535, 549, or 559.
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 4, 16, 28, 40, 52, 64, 76, 88, 100, 112, 124, 136, 148, 160, 172, 184, 196, 208, 220, 232, 247, 259, 271, 283, 295, 307, 319, 331, 343, 355, 367, 380, 391, 403, 415, 427, 439, 451, 463, 475, 487, 499, 511, 523, 535, 549, or 559, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions.
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this paragraph are referred to herein as “variants.”
  • such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 3, a CDR-H2 of SEQ ID NO: , a CDR-H1 of SEQ ID NO: 1, a CDR-L3 of SEQ ID NO: 6, a CDR-L2 of SEQ ID NO: 5, and a CDR-L1 of SEQ ID NO: 4.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 3
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 2
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 1
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 6
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 5
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 3, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 2, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 1, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 6, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 5, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 4, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 15, a CDR-H2 of SEQ ID NO: 14, a CDR-H1 of SEQ ID NO: 13, a CDR-L3 of SEQ ID NO: 18, a CDR-L2 of SEQ ID NO: 17, and a CDR-L1 of SEQ ID NO: 16.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 15
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 14
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 13
  • the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 18
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 17
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 15, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 14, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 13, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 18, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 17, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 16, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 27, a CDR-H2 of SEQ ID NO: 26, a CDR-H1 of SEQ ID NO: 25, a CDR-L3 of SEQ ID NO: 30, a CDR-L2 of SEQ ID NO: 29, and a CDR-L1 of SEQ ID NO: 28.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 27
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 26
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 25
  • the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 30
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 29
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 27, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 26, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 25, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 30, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 29, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 28, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 39, a CDR-H2 of SEQ ID NO: 38, a CDR-H1 of SEQ ID NO: 37, a CDR-L3 of SEQ ID NO: 42, a CDR-L2 of SEQ ID NO: 41, and a CDR-L1 of SEQ ID NO: 40.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 39
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 38
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 37
  • the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 42
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 41
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 39, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 38, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 37, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 42, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 41, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 40, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 51, a CDR-H2 of SEQ ID NO: 50, a CDR-H1 of SEQ ID NO: 49, a CDR-L3 of SEQ ID NO: 53, a CDR-L2 of SEQ ID NO: 52, and a CDR-L1 of SEQ ID NO: 51.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 51
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 50
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 49
  • the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 53
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 52
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 51, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 50, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 49, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 53, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 52, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 51, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 63, a CDR-H2 of SEQ ID NO: 62, a CDR-H1 of SEQ ID NO: 61, a CDR-L3 of SEQ ID NO: 66, a CDR-L2 of SEQ ID NO: 65, and a CDR-L1 of SEQ ID NO: 64.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 63
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 62
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 61
  • the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 66
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 65
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 61, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 62, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 61, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 66, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 65, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 64, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 75, a CDR-H2 of SEQ ID NO: 74, a CDR-H1 of SEQ ID NO: 73, a CDR-L3 of SEQ ID NO: 78, a CDR-L2 of SEQ ID NO: 77, and a CDR-L1 of SEQ ID NO: 76.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 75
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 74
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 73
  • the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 78
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 77
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 75, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 74, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 73, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 78, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 77, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 76, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 87, a CDR-H2 of SEQ ID NO: 86, a CDR-H1 of SEQ ID NO: 85, a CDR-L3 of SEQ ID NO: 90, a CDR-L2 of SEQ ID NO: 89, and a CDR-L1 of SEQ ID NO: 88.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 87
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 86
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 85
  • the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 90
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 89
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a C
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 87, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 86, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 85, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 90, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 89, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 88, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 99, a CDR-H2 of SEQ ID NO: 98, a CDR-H1 of SEQ ID NO: 97, a CDR-L3 of SEQ ID NO: 102, a CDR-L2 of SEQ ID NO: 101, and a CDR-L1 of SEQ ID NO: 100.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 99
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 98
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 97
  • the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 102
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 101
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a C
  • the CORED is a CDR-H3 of SEQ ID NO: 99, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 98, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 97, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 102, with up to 1, 2,
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 101, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 100, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 111, a CDR-H2 of SEQ ID NO: 110, a CDR-H1 of SEQ ID NO: 109, a CDR-L3 of SEQ ID NO: 114, a CDR-L2 of SEQ ID NO: 113, and a CDR-L1 of SEQ ID NO: 112.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 111
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 110
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 109
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 114
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 113
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 111, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 110, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 109, with up to 1, 2, 3,
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 114, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 113, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 112, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 123, a CDR-H2 of SEQ ID NO: 122, a CDR-H1 of SEQ ID NO: 121, a CDR-L3 of SEQ ID NO: 126, a CDR-L2 of SEQ ID NO: 125, and a CDR-L1 of SEQ ID NO: 124.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 123
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 122
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 121
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 126
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 125
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 123, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 122, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 121, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 126, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 125, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 124, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 135, a CDR-H2 of SEQ ID NO: 134, a CDR-H1 of SEQ ID NO: 133, a CDR-L3 of SEQ ID NO: 138, a CDR-L2 of SEQ ID NO: 137, and a CDR-L1 of SEQ ID NO: 136.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 135, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 134, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 133, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 136, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 135, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 135, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 134, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 133, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 136, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 135, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 134, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 148, a CDR-H2 of SEQ ID NO: 147, a CDR-H1 of SEQ ID NO: 146, a CDR-L3 of SEQ ID NO: 151, a CDR-L2 of SEQ ID NO: 150, and a CDR-L1 of SEQ ID NO: 149.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 148
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 147
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 146
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 151
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 150
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 148, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 147, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 146, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 151, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 150, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 149, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 159, a CDR-H2 of SEQ ID NO: 158, a CDR-H1 of SEQ ID NO: 157, a CDR-L3 of SEQ ID NO: 162, a CDR-L2 of SEQ ID NO: 161, and a CDR-L1 of SEQ ID NO: 160.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 159
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 158
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 157
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 162
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 161
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 159, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 158, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 157, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 162, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 161, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 160, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 171, a CDR-H2 of SEQ ID NO: 170, a CDR-H1 of SEQ ID NO: 169, a CDR-L3 of SEQ ID NO: 174, a CDR-L2 of SEQ ID NO: 173, and a CDR-L1 of SEQ ID NO: 172.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 171
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 170
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 169
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 174
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 173
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 171, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 170, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 169, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 174, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 173, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 172, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 183, a CDR-H2 of SEQ ID NO: 182, a CDR-H1 of SEQ ID NO: 181, a CDR-L3 of SEQ ID NO: 186, a CDR-L2 of SEQ ID NO: 185, and a CDR-L1 of SEQ ID NO: 184.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 183
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 182
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 181
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 186
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 185
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 183, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 182, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 181, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 186, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 185, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 184, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 195, a CDR-H2 of SEQ ID NO: 194, a CDR-H1 of SEQ ID NO: 193, a CDR-L3 of SEQ ID NO: 198, a CDR-L2 of SEQ ID NO: 197, and a CDR-L1 of SEQ ID NO: 196.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 195
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 194
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 193
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 198
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 197
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 195, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 194, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 193, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 198, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 197, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 196, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 207, a CDR-H2 of SEQ ID NO: 206, a CDR-H1 of SEQ ID NO: 205, a CDR-L3 of SEQ ID NO: 210, a CDR-L2 of SEQ ID NO: 209, and a CDR-L1 of SEQ ID NO: 208.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 207
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 206
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 205
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 210
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 209
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 207, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 206, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 205, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 210, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 209, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 208, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 219, a CDR-H2 of SEQ ID NO: 218, a CDR-H1 of SEQ ID NO: 217, a CDR-L3 of SEQ ID NO: 222, a CDR-L2 of SEQ ID NO: 221, and a CDR-L1 of SEQ ID NO: 220.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 219
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 21
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 217
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 222
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 221
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 219, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 218, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 217, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 222, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 221, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 220, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 231, a CDR-H2 of SEQ ID NO: 230, a CDR-H1 of SEQ ID NO: 229, a CDR-L3 of SEQ ID NO: 234, a CDR-L2 of SEQ ID NO: 233, and a CDR-L1 of SEQ ID NO: 232.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 231
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 230
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 229
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 234
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 233
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 231, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 230, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 229, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 234, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 233, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 232, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 246, a CDR-H2 of SEQ ID NO: 245, a CDR-H1 of SEQ ID NO: 244, a CDR-L3 of SEQ ID NO: 249, a CDR-L2 of SEQ ID NO: 248, and a CDR-L1 of SEQ ID NO: 247.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 246, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 245, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 244, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 249, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 248, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 246, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 245, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 244, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 249, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 248, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 247, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 258, a CDR-H2 of SEQ ID NO: 257, a CDR-H1 of SEQ ID NO: 256, a CDR-L3 of SEQ ID NO: 261, a CDR-L2 of SEQ ID NO: 260, and a CDR-L1 of SEQ ID NO: 259.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 258, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 257, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 256, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 261, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 260, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 258, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 257, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 256, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 261, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 260, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 259, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 270, a CDR-H2 of SEQ ID NO: 269, a CDR-H1 of SEQ ID NO: 268, a CDR-L3 of SEQ ID NO: 273, a CDR-L2 of SEQ ID NO: 272, and a CDR-L1 of SEQ ID NO: 271.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 270
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 269
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 268
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 273
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 272
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 270, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 269, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 268, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 273, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 272, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 271, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 282, a CDR-H2 of SEQ ID NO: 281, a CDR-H1 of SEQ ID NO: 280, a CDR-L3 of SEQ ID NO: 285, a CDR-L2 of SEQ ID NO: 284, and a CDR-L1 of SEQ ID NO: 283.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 282
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 281
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 280
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 285
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 284
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 282, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 281, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 280, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 285, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 284, with up to 1, 2, 3, or 4 amino acid substitutions; and
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 283, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 294, a CDR-H2 of SEQ ID NO: 293, a CDR-H1 of SEQ ID NO: 292, a CDR-L3 of SEQ ID NO: 297, a CDR-L2 of SEQ ID NO: 296, and a CDR-L1 of SEQ ID NO: 295.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 294, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 293, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 292, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 297, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 296, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 294, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 293, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 292, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 297, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 296, with up to 1, 2, 3, or 4 amino acid substitutions; and
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 295, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 306, a CDR-H2 of SEQ ID NO: 305, a CDR-H1 of SEQ ID NO: 304, a CDR-L3 of SEQ ID NO: 309, a CDR-L2 of SEQ ID NO: 308, and a CDR-L1 of SEQ ID NO: 307.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 306
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 305
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 304
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 309
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 308
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 306, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 305, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 304, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 309, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 308, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 307, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 318, a CDR-H2 of SEQ ID NO: 317, a CDR-H1 of SEQ ID NO: 316, a CDR-L3 of SEQ ID NO: 321, a CDR-L2 of SEQ ID NO: 320, and a CDR-L1 of SEQ ID NO: 319.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 318
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 317
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 316
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 321
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 320
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 318, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 317, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 316, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 321, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 320, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 319, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 330, a CDR-H2 of SEQ ID NO: 329, a CDR-H1 of SEQ ID NO: 328, a CDR-L3 of SEQ ID NO: 333, a CDR-L2 of SEQ ID NO: 332, and a CDR-Ll of SEQ ID NO: 331.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 330
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 329
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 330
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 333
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 332
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 330, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 329, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 328, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 333, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 332, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 331, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 342, a CDR-H2 of SEQ ID NO: 341, a CDR-H1 of SEQ ID NO: 340, a CDR-L3 of SEQ ID NO: 345, a CDR-L2 of SEQ ID NO: 344, and a CDR-L1 of SEQ ID NO: 343.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 342
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 341
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 340
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 345
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 344
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 342, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 341, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 340, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 345, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 344, with up to 1, 2, 3, or 4 amino acid substitutions;
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 343, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 354, a CDR-H2 of SEQ ID NO: 353, a CDR-H1 of SEQ ID NO: 352, a CDR-L3 of SEQ ID NO: 357, a CDR-L2 of SEQ ID NO: 356, and a CDR-L1 of SEQ ID NO: 355.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 354, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 353, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 352, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 357, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 356, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 354, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 353, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 352, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 357, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 356, with up to 1, 2, 3, or 4 amino acid substitutions; and
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 355, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 366, a CDR-H2 of SEQ ID NO: 365, a CDR-H1 of SEQ ID NO: 364, a CDR-L3 of SEQ ID NO: 369, a CDR-L2 of SEQ ID NO: 368, and a CDR-L1 of SEQ ID NO: 367.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 366
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 365
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 364
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 369
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 368
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 366, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 365, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 364, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 369, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 368, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 367, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 378, a CDR-H2 of SEQ ID NO: 377, a CDR-H1 of SEQ ID NO: 376, a CDR-L3 of SEQ ID NO: 381, a CDR-L2 of SEQ ID NO: 380, and a CDR-L1 of SEQ ID NO: 379.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 378
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 377
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 376
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 381
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 380
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 378, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 377, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 376, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 381, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 380, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 379, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 390, a CDR-H2 of SEQ ID NO: 389, a CDR-H1 of SEQ ID NO: 388, a CDR-L3 of SEQ ID NO: 393, a CDR-L2 of SEQ ID NO: 392, and a CDR-L1 of SEQ ID NO: 391.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 390
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 389
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 388
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 393
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 392
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 390, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 389, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 388, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 393, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 392, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 391, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 402, a CDR-H2 of SEQ ID NO: 401, a CDR-H1 of SEQ ID NO: 400, a CDR-L3 of SEQ ID NO: 405, a CDR-L2 of SEQ ID NO: 404, and a CDR-L1 of SEQ ID NO: 403.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 402
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 401
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 400
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 405
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 404
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 402, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 401, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 400, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 405, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 404, with up to 1, 2, 3, or 4 amino acid substitutions; and
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 403, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 414, a CDR-H2 of SEQ ID NO: 413, a CDR-H1 of SEQ ID NO: 412, a CDR-L3 of SEQ ID NO: 417, a CDR-L2 of SEQ ID NO: 416, and a CDR-L1 of SEQ ID NO: 415.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 414
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 413
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 412
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 417
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 416
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 414, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 413, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 412, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 417, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 416, with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 415, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 426, a CDR-H2 of SEQ ID NO: 425, a CDR-H1 of SEQ ID NO: 424, a CDR-L3 of SEQ ID NO: 429, a CDR-L2 of SEQ ID NO: 428, and a CDR-L1 of SEQ ID NO: 427.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 426
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 425
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 424
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 429
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 428
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 426, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 425, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 424, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 429, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 428 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 427, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 438, a CDR-H2 of SEQ ID NO: 437, a CDR-H1 of SEQ ID NO: 436, a CDR-L3 of SEQ ID NO: 441, a CDR-L2 of SEQ ID NO: 440, and a CDR-L1 of SEQ ID NO: 439.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 438
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 437
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 436
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 441
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 440
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 438, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 437, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 436, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 441, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 440 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 439, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 450, a CDR-H2 of SEQ ID NO: 449, a CDR-H1 of SEQ ID NO: 448 a CDR-L3 of SEQ ID NO: 453, a CDR-L2 of SEQ ID NO: 452, and a CDR-L1 of SEQ ID NO: 451.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 450
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 449
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 448
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 453
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 452
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 450, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 449, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 448, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 453, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 452 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 451, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 462, a CDR-H2 of SEQ ID NO: 461, a CDR-H1 of SEQ ID NO: 460 a CDR-L3 of SEQ ID NO: 465, a CDR-L2 of SEQ ID NO: 464, and a CDR-L1 of SEQ ID NO: 463.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 462
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 461
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 460
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 465
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 464
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 462, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 461, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions;
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 460, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 465, with up to 1, 2, 3, 4, or 5 amino acid substitutions;
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 464 with up to 1, 2, 3, or 4 amino acid substitutions; and
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 463, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 474, a CDR-H2 of SEQ ID NO: 473, a CDR-H1 of SEQ ID NO: 472 a CDR-L3 of SEQ ID NO: 477, a CDR-L2 of SEQ ID NO: 476, and a CDR-L1 of SEQ ID NO: 475.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 474
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 473
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 472
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 477
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 476
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 474, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 473, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 472, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 477, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 476 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 475, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 486, a CDR-H2 of SEQ ID NO: 485, a CDR-H1 of SEQ ID NO: 484 a CDR-L3 of SEQ ID NO: 489, a CDR-L2 of SEQ ID NO: 488, and a CDR-L1 of SEQ ID NO: 487.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 486, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 485, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 484, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 489, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 488, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 486, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 485, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 484, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 489, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 488 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 487, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 498, a CDR-H2 of SEQ ID NO: 497, a CDR-H1 of SEQ ID NO: 496 a CDR-L3 of SEQ ID NO: 501, a CDR-L2 of SEQ ID NO: 500, and a CDR-L1 of SEQ ID NO: 499.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 498
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 497
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 496
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 501
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 500
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 498, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 497, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 496, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 501, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 500 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 499, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 510, a CDR-H2 of SEQ ID NO: 509, a CDR-H1 of SEQ ID NO: 508 a CDR-L3 of SEQ ID NO: 513, a CDR-L2 of SEQ ID NO: 512, and a CDR-L1 of SEQ ID NO: 511.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 510
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 509
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 508
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 513
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 512
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 510, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 509, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 508, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 513, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 512 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 511, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 522, a CDR-H2 of SEQ ID NO: 521, a CDR-H1 of SEQ ID NO: 520 a CDR-L3 of SEQ ID NO: 525, a CDR-L2 of SEQ ID NO: 524, and a CDR-L1 of SEQ ID NO: 523.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 522
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 521
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 520
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 525
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 524
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 522, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 521, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 520, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 525, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 524 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 523, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 534, a CDR-H2 of SEQ ID NO: 533, a CDR-H1 of SEQ ID NO: 532 a CDR-L3 of SEQ ID NO: 537, a CDR-L2 of SEQ ID NO: 536, and a CDR-L1 of SEQ ID NO: 535.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 534
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 533
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 532
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 537
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 536
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 534, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 533, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 532, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 537, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 536 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 535, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 546, a CDR-H2 of SEQ ID NO: 545, a CDR-H1 of SEQ ID NO: 544 a CDR-L3 of SEQ ID NO: 549, a CDR-L2 of SEQ ID NO: 548, and a CDR-L1 of SEQ ID NO: 547.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 546
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 545
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 544
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 549
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 548
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 546, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 545, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 544, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 549, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 548 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 547, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 558, a CDR-H2 of SEQ ID NO: 557, a CDR-H1 of SEQ ID NO: 556 a CDR-L3 of SEQ ID NO: 561, a CDR-L2 of SEQ ID NO: 560, and a CDR-L1 of SEQ ID NO: 559.
  • the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 558
  • the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 557
  • the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 556
  • the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 561
  • the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 560
  • the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with
  • the CDR-H3 is a CDR-H3 of SEQ ID NO: 558, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H2 is a CDR-H2 of SEQ ID NO: 557, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions
  • the CDR-H1 is a CDR-H1 of SEQ ID NO: 556, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L3 is a CDR-L3 of SEQ ID NO: 561, with up to 1, 2, 3, 4, or 5 amino acid substitutions
  • the CDR-L2 is a CDR-L2 of SEQ ID NO: 560 with up to 1, 2, 3, or 4 amino acid substitutions
  • the CDR-L1 is a CDR-L1 of SEQ ID NO: 559, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
  • the amino acid substitutions are conservative amino acid substitutions.
  • the antibodies described in this disclosure are referred to herein as “variants” or “clones”.
  • such variants or clones are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein.
  • such variants or cones are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
  • Fc regions of various immunoglobulins are known in the art. See Schroeder and Cavacini, J. Allergy Clin. Immunol., 2010, 125:S41-52, incorporated by reference in its entirety.
  • the Fc region may be a naturally occurring Fc region, or an Fc region modified as described in the art or elsewhere in this disclosure.
  • Fc polypeptide of a dimeric Fc as used herein refers to one of the two polypeptides forming the dimeric Fc domain, i.e. a polypeptide comprising C-terminal constant regions of an immunoglobulin heavy chain, capable of stable self-association.
  • an Fc polypeptide of a dimeric IgG Fc comprises an IgG CH2 and an IgG CH3 constant domain sequence.
  • An Fc can be of the class IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2.
  • Fc receptor and “FcR” are used to describe a receptor that binds to the Fc region of an antibody.
  • an FcR can be a native sequence human FcR.
  • an FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the FcyRI, FcyRII, and FcyRIII subclasses, including allelic variants and alternatively spliced forms of these receptors.
  • FcyRII receptors include FcyRIIA (an “activating receptor”) and FcyRIIB (an “inhibiting receptor”), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof.
  • Immunoglobulins of other isotypes can also be bound by certain FcRs (see, e.g., Janeway et al., Immuno Biology: the immune system in health and disease, (Elsevier Science Ltd., NY) (4th ed., 1999)).
  • Activating receptor FcyRIIA contains an immunoreceptor tyrosine-based activation motif (IT AM) in its cytoplasmic domain.
  • Inhibiting receptor FcyRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITEM) in its cytoplasmic domain (reviewed in Daeron, Annu. Rev. Immunol. 15:203-234 (1997)).
  • FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995).
  • FcR FcR
  • the term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976); and Kim et al., J. Immunol. 24:249 (1994)).
  • FcRn neonatal receptor
  • Modifications in the CH2 domain can affect the binding of FcRs to the Fc.
  • a number of amino acid modifications in the Fc region are known in the art for selectively altering the affinity of the Fc for different Fcgamma receptors.
  • the Fc comprises one or more modifications to promote selective binding of Fc-gamma receptors.
  • F243L (Stewart R, Thom G, Levens M, et al. Protein Eng Des Sei. 2011 Sep;24(9):671-8.), S298A/E333A/K334A (Shields RL, Namenuk AK, Hong K, et al. J Biol Chem. 2001 Mar 2;276(9):6591-604);
  • an antibody described herein includes modifications to improve its ability to mediate effector function.
  • modifications are known in the art and include afucosylation, or engineering of the affinity of the Fc towards an activating receptor, mainly FCGR3a for ADCC, and towards Clq for CDC.
  • FCGR3a for ADCC
  • Clq for CDC.
  • Table B summarizes various designs reported in the literature for effector function engineering.
  • the GlymaX® technology (ProBioGen AG) is based on the introduction of a gene for an enzyme which deflects the cellular pathway of fucose biosynthesis into cells used for antibody production. This prevents the addition of the sugar “fucose” to the N-linked antibody carbohydrate part by antibody-producing cells, (von Horsten et al. (2010) Glycobiology. 2010 Dec; 20 (12): 1607-18. Another approach to obtaining antibodies with lowered levels of fucosylation can be found in U.S.
  • patent 8,409,572 which teaches selecting cell lines for antibody production for their ability to yield lower levels of fucosylation on antibodies can be fully afucosylated (meaning they contain no detectable fucose) or they can be partially afucosylated, meaning that the isolated antibody contains less than 95%, less than 85%, less than 75%, less than 65%, less than 55%, less than 45%, less than 35%, less than 25%, less than 15% or less than 5% of the amount of fucose normally detected for a similar antibody produced by a mammalian expression system.
  • an antibody described herein can include a dimeric Fc that comprises one or more amino acid modifications as noted in Table B that confer improved effector function.
  • the antibody can be afucosylated to improve effector function.
  • Examples of cell lines capable of producing defucosylated antibody include CHO-DG44 with stable overexpression of the bacterial oxidoreductase GDP-6-deoxy-D-lyxo-4-hexylose reductase (RMD) (see Henning von Horsten et al., Glycobiol 2010, 20: 1607-1618) or Lecl3 CHO cells, which are deficient in protein fucosylation (see Ripka et al., Arch. Biochem. Biophys., 1986, 249:533-545; U.S. Pat. Pub. No.
  • RMD bacterial oxidoreductase GDP-6-deoxy-D-lyxo-4-hexylose reductase
  • Examples of cell lines capable of producing defucosylated antibody include CHO- DG44 with stable overexpression of the bacterial oxidoreductase GDP-6-deoxy-D-lyxo-4- hexylose reductase (RMD) (see Henning von Horsten et al., Glycobiol 2010, 20: 1607-1618) or Lecl3 CHO cells, which are deficient in protein fucosylation (see Ripka et al., Arch. Biochem. Biophys., 1986, 249:533-545; U.S. Pat. Pub. No.
  • RMD bacterial oxidoreductase GDP-6-deoxy-D-lyxo-4- hexylose reductase
  • knockout cell lines such as alpha- 1,6-fucosyltransf erase gene or FUT8 knockout CHO cells (see Yamane- Ohnuki et al., Biotech. Bioeng., 2004, 87: 614-622; Kanda et al., Biotechnol. Bioeng., 2006, 94:680-688; and WO 2003/085107; each of which is incorporated by reference in its entirety).
  • Antibodies can be fully afucosylated (meaning they contain no detectable fucose) or they can be partially afucosylated, meaning that the isolated antibody contains less than 95%, less than 85%, less than 75%, less than 65%, less than 55%, less than 45%, less than 35%, less than 25%, less than 15% or less than 5% of the amount of fucose normally detected for a similar antibody produced by a mammalian expression system.
  • an antibody provided herein comprises an IgGl domain with reduced fucose content at position Asn 297 compared to a naturally occurring IgGl domain.
  • Fc domains are known to have improved ADCC. See Shields et al., J. Biol. Chem., 2002, 277:26733-26740, incorporated by reference in its entirety.
  • such antibodies do not comprise any fucose at position Asn 297. The amount of fucose may be determined using any suitable method, for example as described in WO 2008/077546, incorporated by reference in its entirety.
  • an antibody provided herein comprises an Fc region with one or more amino acid substitutions which improve ADCC, such as a substitution at one or more of positions 298, 333, and 334 of the Fc region.
  • an antibody provided herein comprises an Fc region with one or more amino acid substitutions at positions 239, 332, and 330, as described in Lazar et al., Proc. Natl. Acad. Sci. USA, 2006,103:4005-4010, incorporated by reference in its entirety.
  • an antibody provided herein comprises an Fc region with at least one galactose residue in the oligosaccharide attached to the Fc region.
  • Such antibody variants may have improved CDC function. Examples of such antibody variants are described, for example, in WO 1997/30087; WO 1998/58964; and WO 1999/22764; each of which his incorporated by reference in its entirety.
  • an antibody provided herein comprises one or more alterations that improves or diminishes Clq binding and/or CDC. See U.S. Pat. No. 6,194,551; WO 99/51642; and Idusogie et al., J. Immunol., 2000, 164:4178-4184; each of which is incorporated by reference in its entirety.
  • the affinity of a molecule X for its partner Y can be represented by the dissociation equilibrium constant (KD).
  • KD dissociation equilibrium constant
  • the kinetic components that contribute to the dissociation equilibrium constant are described in more detail below.
  • Affinity can be measured by common methods known in the art, including those described herein, such as surface plasmon resonance (SPR) technology (e.g., BIACORE®) or biolayer interferometry (e.g., FORTEBIO®).
  • SPR surface plasmon resonance
  • BIACORE® BIACORE®
  • FORTEBIO® biolayer interferometry
  • the terms “bind,” “specific binding,” “specifically binds to,” “specific for,” “selectively binds,” and “selective for” a particular antigen (e.g., a polypeptide target) or an epitope on a particular antigen mean binding that is measurably different from a non-specific or non-selective interaction (e.g., with a non-target molecule).
  • Specific binding can be measured, for example, by measuring binding to a target molecule and comparing it to binding to a non-target molecule.
  • Specific binding can also be determined by competition with a control molecule that mimics the epitope recognized on the target molecule.
  • the affinity of a fibrin antibody for a non-target molecule is less than about 50% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 40% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 30% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 20% of the affinity for fibrin.
  • the affinity of a fibrin antibody for a non-target molecule is less than about 10% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 1% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 0.1% of the affinity for fibrin.
  • the term “competes with” or “cross-competes with” indicates that the two or more antibodies compete for binding to an antigen (e.g., fibrin).
  • fibrin is coated on a surface and contacted with a first fibrin antibody, after which a second fibrin antibody is added.
  • a first fibrin antibody is coated on a surface and contacted with fibrin, and then a second fibrin antibody is added. If the presence of the first fibrin antibody reduces binding of the second fibrin antibody, in either assay, then the antibodies compete with each other.
  • the term “competes with” also includes combinations of antibodies where one antibody reduces binding of another antibody, but where no competition is observed when the antibodies are added in the reverse order.
  • the first and second antibodies inhibit binding of each other, regardless of the order in which they are added.
  • one antibody reduces binding of another antibody to its antigen by at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% as measured in a competitive binding assay.
  • concentrations of the antibodies used in the competition assays based on the affinities of the antibodies for fibrin and the valency of the antibodies.
  • the assays described in this definition are illustrative, and a skilled artisan can utilize any suitable assay to determine if antibodies compete with each other. Suitable assays are described, for example, in Cox et al., “Immunoassay Methods,” in Assay Guidance Manual [Internet], Updated December 24, 2014 (ncbi.nlm.nih.gov/books/NBK92434/; accessed September 29, 2015); Silman et al., Cytometry, 2001, 44:30-37; and Finco et al., J. Pharm. Biomed. Anal., 2011, 54:351-358; each of which is incorporated by reference in its entirety.
  • a test antibody competes with a reference antibody if an excess of a test antibody (e.g., at least 2x, 5x, lOx, 20x, or lOOx) inhibits or blocks binding of the reference antibody by, e.g., at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% as measured in a competitive binding assay.
  • Antibodies identified by competition assay include antibodies binding to the same epitope as the reference antibody and antibodies binding to an adjacent epitope sufficiently proximal to the epitope bound by the reference antibody for steric hindrance to occur.
  • a second, competing antibody can be identified that competes for binding to fibrin with a first antibody described herein.
  • the second antibody can block or inhibit binding of the first antibody by, e.g., at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% as measured in a competitive binding assay.
  • the second antibody can displace the first antibody by greater than 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%.
  • an anti- fibrin antibody does not substantially bind myeloid cells present outside of cancer tissue.
  • an anti- fibrin antibody does not substantially bind stimulatory myeloid cells present in cancer tissue.
  • an anti- fibrin antibody binds to residues y377-395 of the fibrin or fibrinogen yC domain (SEQ ID NO: 31) of human fibrin.
  • the binding epitope includes the residues within the numerical range (e.g., residues 377-395 of fibrin), the beginning residue of each range (e.g., residues 377-394 of human fibrin) and the end residue of each range (e.g., residues 378-395 of human fibrin), or any combination thereof.
  • an antibody provided herein binds human Fibrin with a KD of less than or equal to about 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.090, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 1.95, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, or lO x IO' 6 M, as measured by ELISA or any other suitable method known in the art.
  • the KD of the antibody provided herein for the binding of human fibrin is between about 0.001-0.01, 0.01- 0.1, 0.01-0.05, 0.05-0.1, 0.1-0.5, 0.5-1, 0.25-0.75, 0.25-0.5, 0.5-0.75, 0.75-1, 0.75-2, 1.1-1.2, 1.2-1.3, 1.3-1.4, 1.4-1.5, 1.5-1.6, 1.6-1.7, 1.7-1.8, 1.8-1.9, 1.9-2, 1-2, 1-5, 2-7, 3-8, 3-5, 4-6, 5-7, 6-8, 7-9, 7-10, or 5-10xl0' 6 M, as measured by ELISA or any other suitable method known in the art .
  • an antibody provided herein binds human Fibrin with a KD of less than or equal to about 1 x 10-8 M, or less than or equal to above 1 x 10' 9 M as measured by ELISA or any other suitable method known in the art.
  • the antibody provided herein binds human fibrin with a KD of less than or equal to about 10, 9, 8, 7, 6, 5, 4.5, 4, 3.5, 3, 2.5, 2, 1.98, 1.95, 1.9, 1.85, 1.8, 1.75, 1.7, 1.65, 1.6, 1.55, 1.50, 1.45, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.85, 0.8, 0.75, 0.7, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15, 0.1, 0.05, 0.01, 0.005, 0.001, 0.0005, or 0.0001 x 10' 8 M, or less, as measured by ELISA or any other suitable method known in the art .
  • the antibody provided herein binds human fibrin with a KD between 5-3, 4-2, 3-1, 1.9-1.8, 1.8-1.7, 1.7-1.6, 1.6-1.5, 1.9-1.5, 1.5-1, 1-0.8, 1-0.5, 0.9-0.6, 0.7-0.4, 0.6- 0.2, 0.5-0.3, 0.3-0.2, 0.2-0.1, 0.1-0.01, 0.01-0.001, or 0.001-0.0001 x IO' 8 M as measured by ELISA or any other suitable method known in the art.
  • an antibody provided herein binds human FGG with a KD of less than or equal to about 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, 0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 1.95, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, or 10 x 10' 6 M, as measured by ELISA or any other suitable method known in the art.
  • the KD of the antibody provided herein for the binding of human FGG is between about 0.0001-0.001, 0.0001-0.0005, 0.0005-0.001, 0.001-0.01, 0.01-0.1, 0.01-0.05, 0.05-0.1, 0.1-0.5, 0.5-1, 0.25-0.75, 0.25-0.5, 0.5-0.75, 0.75-1, 0.75-2, 1.1-1.2, 1.2-1.3, 1.3- 1.4, 1.4-1.5, 1.5-1.6, 1.6-1.7, 1.7-1.8, 1.8-1.9, 1.9-2, 1-2, 1-5, 2-7, 3-8, 3-5, 4-6, 5-7, 6-8, 7-9, 7-10, or 5-10 x 10' 8 M, as measured by ELISA or any other suitable method known in the art .
  • an antibody provided herein binds human FGG with a KD of less than or equal to about 1 x 10-8 M, or less than or equal to above 1 x 10-9 M as measured by ELISA or any other suitable method known in the art.
  • an antibody provided herein binds P2 peptide with a KD of less than or equal to about 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, 0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 1.95, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, or 10 x 10' 6 M, as measured by ELISA or any other suitable method known in the art.
  • the KD of the antibody provided herein for the binding of P2 peptide is between about 0.0001-0.001, 0.0001-0.0005, 0.001-0.01, 0.01-0.1, 0.01-0.05, 0.05-0.1, 0.1- 0.5, 0.5-1, 0.25-0.75, 0.25-0.5, 0.5-0.75, 0.75-1, 0.75-2, 1.1-1.2, 1.2-1.3, 1.3-1.4, 1.4-1.5, 1.5- 1.6, 1.6-1.7, 1.7-1.8, 1.8-1.9, 1.9-2, 1-2, 1-5, 2-7, 3-8, 3-5, 4-6, 5-7, 6-8, 7-9, 7-10, or 5-10 x 10' 6 M, as measured by ELISA or any other suitable method known in the art .
  • an antibody provided herein binds P2 peptide with a KD of less than or equal to about 1 x 10' 8 M, or less than or equal to above 1 x 10' 9 M as measured by ELISA or any other suitable method known in the art.
  • an antibody provided herein binds fibrinogen with a KD of greater than or equal to about 1 x 10' 6 M, greater than or equal to above 1 x 10' 5 M, greater than or equal to above 1 x 10' 4 M, or greater than or equal to above 1 x 10' 3 M as measured by ELISA or any other suitable method known in the art.
  • the ratio of KD of the binding affinity of the antibody described herein to fibrin compared to fibrinogen is equal to or greater than 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 15, 20, 25,30, 35, 40, 45,50, 60, 70, 80, 90, 100, 150, 200, 300, 400, 500, or 1000.
  • “Effector functions” refer to those biological activities mediated by the Fc region of an antibody, which activities may vary depending on the antibody isotype.
  • Examples of antibody effector functions include receptor ligand blocking, agonism, or antagonism, Clq binding to activate complement dependent cytotoxicity (CDC), Fc receptor binding to activate antibody-dependent cellular cytotoxicity (ADCC), and antibody dependent cellular phagocytosis (ADCP).
  • the effector function of the fibrin antibody described herein is antagonism and blocks Mac-1 receptor binding to fibrin.
  • compositions comprising the antibodies including pharmaceutical compositions comprising any one or more of the antibodies described herein with one or more pharmaceutically acceptable excipients.
  • the composition is sterile.
  • the pharmaceutical compositions generally comprise an effective amount of an antibody.
  • compositions can comprise, in addition to one or more of the antibodies disclosed herein, a pharmaceutically acceptable excipient, carrier, buffer, stabilizer or other materials well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient.
  • a pharmaceutically acceptable excipient e.g. oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, intraperitoneal routes.
  • compositions for oral administration can be in tablet, capsule, powder or liquid form.
  • a tablet can include a solid carrier such as gelatin or an adjuvant.
  • Liquid pharmaceutical compositions generally include a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil. Physiological saline solution, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol can be included.
  • the active ingredient will be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability.
  • a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability.
  • isotonic vehicles such as Sodium Chloride Injection, Ringer's Injection, Lactated Ringer's Injection.
  • Preservatives, stabilisers, buffers, antioxidants and/or other additives can be included, as required.
  • the anti-fibrin antibody that is to be given to an individual, administration is preferably in a “therapeutically effective amount” or “prophylactically effective amount” (as the case can be, although prophylaxis can be considered therapy), this being sufficient to show benefit to the individual.
  • a “therapeutically effective amount” or “prophylactically effective amount” as the case can be, although prophylaxis can be considered therapy
  • the actual amount administered, and rate and time-course of administration will depend on the nature and severity of protein aggregation disease being treated. Prescription of treatment, e.g. decisions on dosage etc., is within the responsibility of general practitioners and other medical doctors, and typically takes account of the disorder to be treated, the condition of the individual patient, the site of delivery, the method of administration and other factors known to practitioners. Examples of the techniques and protocols mentioned above can be found in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. (ed), 1980.
  • a composition can be administered alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated.
  • Antibodies described herein can be produced using recombinant methods and compositions, e.g., as described in U.S. Pat. No. 4,816,567.
  • isolated nucleic acid encoding an antibody described herein is provided.
  • Such nucleic acid may encode an amino acid sequence comprising the VL and/or an amino acid sequence comprising the VH of the antibody (e.g., the light and/or heavy chains of the antibody) or an amino acid sequence comprising the VHH of a single domain antibody.
  • one or more vectors e.g., expression vectors
  • the nucleic acid is provided in a multi ci stronic vector.
  • a host cell comprising such nucleic acid.
  • a host cell comprises (e.g., has been transformed with): (1) a vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antibody and an amino acid sequence comprising the VH of the antigen-binding polypeptide construct, or (2) a first vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antigen-binding polypeptide construct and a second vector comprising a nucleic acid that encodes an amino acid sequence comprising the VH of the antigen-binding polypeptide construct.
  • the host cell is eukaryotic, e.g.
  • a method of making an antibody comprises culturing a host cell comprising nucleic acid encoding the antibody, as provided above, under conditions suitable for expression of the antibody, and optionally recovering the antibody from the host cell (or host cell culture medium).
  • nucleic acid encoding an antibody is isolated and inserted into one or more vectors for further cloning and/or expression in a host cell.
  • nucleic acid may be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the antibody).
  • the protein in certain embodiments is present at about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% or less of the dry weight of the cells.
  • the protein in certain embodiments, is present in the culture medium at about 5 g/L, about 4 g/L, about 3 g/L, about 2 g/L, about 1 g/L, about 750 mg/L, about 500 mg/L, about 250 mg/L, about 100 mg/L, about 50 mg/L, about 10 mg/L, or about 1 mg/L or less of the dry weight of the cells.
  • “substantially purified” heteromultimer produced by the methods described herein has a purity level of at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, specifically, a purity level of at least about 75%, 80%, 85%, and more specifically, a purity level of at least about 90%, a purity level of at least about 95%, a purity level of at least about 99% or greater as determined by appropriate methods such as SDS/PAGE analysis, RP-HPLC, SEC, and capillary electrophoresis.
  • Suitable host cells for cloning or expression of antibody-encoding vectors include prokaryotic or eukaryotic cells described herein.
  • Recombinant host cells or host cells are cells that include an exogenous polynucleotide, regardless of the method used for insertion, for example, direct uptake, transduction, f-mating, or other methods known in the art to create recombinant host cells.
  • the exogenous polynucleotide may be maintained as a nonintegrated vector, for example, a plasmid, or alternatively, may be integrated into the host genome.
  • Host cells can include CHO, derivatives of CHO, NSO, Sp2O, CV-1, VERO-76, HeLa, HepG2, Per.C6, or BHK.
  • antibody may be produced in bacteria, in particular when glycosylation and Fc effector function are not needed.
  • expression of antibody fragments and polypeptides in bacteria see, e.g., U.S. Pat. Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, N.J., 2003), pp. 245-254, describing expression of antibody fragments in E. coli.)
  • the antibody may be isolated from the bacterial cell paste in a soluble fraction and can be further purified.
  • eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for antibody-encoding vectors, including fungi and yeast strains whose glycosylation pathways have been “humanized,” resulting in the production of an antibody with a partially or fully human glycosylation pattern. See Gerngross, Nat. Biotech. 22: 1409-1414 (2004), and Li et al., Nat. Biotech. 24:210-215 (2006).
  • Suitable host cells for the expression of glycosylated antibodies are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains have been identified which may be used in conjunction with insect cells, particularly for transfection of Spodoptera frugiperda cells.
  • Plant cell cultures can also be utilized as hosts. See, e.g., U.S. Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing PLANTIBODIESTM technology for producing antibodies in transgenic plants).
  • Vertebrate cells may also be used as hosts.
  • mammalian cell lines that are adapted to grow in suspension may be useful.
  • useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, e.g., in Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells as described, e.g., in Mather, Biol. Reprod.
  • monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK; buffalo rat liver cells (BRL 3 A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described, e.g., in Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells.
  • Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci.
  • the antibodies described herein are produced in stable mammalian cells, by a method comprising: transfecting at least one stable mammalian cell with: nucleic acid encoding the antibody, in a predetermined ratio; and expressing the nucleic acid in the at least one mammalian cell.
  • the predetermined ratio of nucleic acid is determined in transient transfection experiments to determine the relative ratio of input nucleic acids that results in the highest percentage of the antibody in the expressed product.
  • the method of producing a glycosylated antibody in stable mammalian cells described herein comprising identifying and purifying the desired glycosylated antibody.
  • the said identification is by one or both of liquid chromatography and mass spectrometry.
  • the antibodies can be purified or isolated after expression.
  • Proteins may be isolated or purified in a variety of ways known to those skilled in the art. Standard purification methods include chromatographic techniques, including ion exchange, hydrophobic interaction, affinity, sizing or gel filtration, and reversed-phase, carried out at atmospheric pressure or at high pressure using systems such as FPLC and HPLC. Purification methods also include electrophoretic, immunological, precipitation, dialysis, and chromatofocusing techniques. Ultrafiltration and diafiltration techniques, in conjunction with protein concentration, are also useful. As is well known in the art, a variety of natural proteins bind Fc and antibodies, and these proteins can find use in the present invention for purification of antibodies.
  • the bacterial proteins A and G bind to the Fc region.
  • the bacterial protein L binds to the Fab region of some antibodies.
  • Purification can often be enabled by a particular fusion partner.
  • antibodies may be purified using glutathione resin if a GST fusion is employed, Ni+2 affinity chromatography if a His-tag is employed or immobilized anti-flag antibody if a flag-tag is used.
  • suitable purification techniques see, e.g. incorporated entirely by reference Protein Purification: Principles and Practice, 3rd Ed., Scopes, Springer-Verlag, NY, 1994, incorporated entirely by reference. The degree of purification necessary will vary depending on the use of the antibodies. In some instances, no purification is necessary.
  • the antibodies are purified using Anion Exchange Chromatography including, but not limited to, chromatography on Q-sepharose, DEAE sepharose, poros HQ, poros DEAF, Toyopearl Q, Toyopearl QAE, Toyopearl DEAE, Resource/Source Q and DEAE, Fractogel Q and DEAE columns.
  • Anion Exchange Chromatography including, but not limited to, chromatography on Q-sepharose, DEAE sepharose, poros HQ, poros DEAF, Toyopearl Q, Toyopearl QAE, Toyopearl DEAE, Resource/Source Q and DEAE, Fractogel Q and DEAE columns.
  • the proteins described herein are purified using Cation Exchange Chromatography including, but not limited to, SP-sepharose, CM sepharose, poros HS, poros CM, Toyopearl SP, Toyopearl CM, Resource/Source S and CM, Fractogel S and CM columns and their equivalents and comparables.
  • antibodies described herein can be chemically synthesized using techniques known in the art (e.g., see Creighton, 1983, Proteins: Structures and Molecular Principles, W. H. Freeman & Co., N.Y and Hunkapiller et al., Nature, 310: 105-111 (1984)).
  • a polypeptide corresponding to a fragment of a polypeptide can be synthesized by use of a peptide synthesizer.
  • nonclassical amino acids or chemical amino acid analogs can be introduced as a substitution or addition into the polypeptide sequence.
  • Non-classical amino acids include, but are not limited to, to the D-isomers of the common amino acids, 2,4diaminobutyric acid, alpha-amino isobutyric acid, 4aminobutyric acid, Abu, 2-amino butyric acid, g-Abu, e-Ahx, 6amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, alanine, fluoro-amino acids, designer amino acids such as methyl amino acids, C-methyl amino acids, N-methyl amino acids, and amino acid analogs in general. Furthermore, the amino acid can be D (d
  • the present application provides methods of contacting fibrin with an anti-fibrin antibody, such as a human or humanized antibody, which results in inhibition of microglial adhesion to the fibrin or fibrinogen yC domain.
  • an anti-fibrin antibody such as a human or humanized antibody
  • the present application provides methods of using the isolated antifibrin antibodies described herein for treatment of a degenerative disorder of the nervous system.
  • described herein is a method for treating a degenerative disorder of the nervous system, the method comprising administering to a mammalian subject a therapeutically effective amount of an anti-fibrin antibody or pharmaceutical composition comprising an anti-fibrin antibody described herein.
  • the present application provides methods of treating a degenerative disorder of the nervous system selected from the group consisting of: multiple sclerosis, spinal cord injury, stroke, and Alzheimer’s Disease.
  • described herein are methods for treating a pathology associated with Mac-1 binding to fibrin or Mac-1 binding with fibrinogen, the method comprising administering to a mammalian subject a therapeutically effective amount an isolated anti- fibrin antibody or a pharmaceutical composition comprising an isolated anti-fibrin antibody described herein.
  • described herein are methods of inhibiting microglia activation, the method comprising administering to a mammalian subject a therapeutically effective amount an isolated anti-fibrin antibody or a pharmaceutical composition comprising an isolated antibody described herein.
  • a method of preventing a degenerative disorder of the nervous system comprising administering to a mammalian subject a therapeutically effective amount an isolated anti-fibrin antibody or a pharmaceutical composition comprising an isolated anti-fibrin antibody described herein.
  • the present application provides methods of preventing a degenerative disorder of the nervous system selected from the group consisting of: multiple sclerosis, spinal cord injury, stroke, and Alzheimer’s Disease.
  • the methods provided herein are useful for the treatment of a degenerative nervous system disorder in an individual.
  • the individual is a human and the antibody is a fibrin antibody described herein.
  • an antibody is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally.
  • An effective amount of an anti-fibrin antibody may be administered for the treatment of cancer.
  • the appropriate dosage of the anti-fibrin antibody may be determined based on the type of cancer to be treated, the type of the anti-fibrin antibody, the severity and course of the cancer, the clinical condition of the individual, the individual’s clinical history and response to the treatment, and the discretion of the attending physician.
  • an antibody provided herein is administered with at least one additional therapeutic agent.
  • Any suitable additional therapeutic or immunotherapeutic agent may be administered with an antibody provided herein.
  • Additional therapeutic agents include agents that are used to treat or prevent a degenerative disorder of the nervous system selected from the group consisting of: multiple sclerosis, spinal cord injury, stroke, and Alzheimer’s Disease.
  • the additional therapeutic agent can be administered by any suitable means.
  • an antibody provided herein and the additional therapeutic agent are included in the same pharmaceutical composition.
  • an antibody provided herein and the additional therapeutic agent are included in different pharmaceutical compositions.
  • administration of the antibody can occur prior to, simultaneously, and/or following, administration of the additional therapeutic agent.
  • administration of an antibody provided herein and the additional therapeutic agent occur within about one month of each other.
  • administration of an antibody provided herein and the additional therapeutic agent occur within about one week of each other.
  • administration of an antibody provided herein and the additional therapeutic agent occur within about one day of each other.
  • administration of an antibody provided herein and the additional therapeutic agent occur within about twelve hours of each other.
  • administration of an antibody provided herein and the additional therapeutic agent occur within about one hour of each other.
  • kits comprising any one or more of the antibody compositions described herein.
  • the kits further contain a component selected from any of secondary antibodies, reagents for immunohistochemistry analysis, pharmaceutically acceptable excipient and instruction manual and any combination thereof.
  • the kit comprises a pharmaceutical composition comprising any one or more of the antibody compositions described herein, with one or more pharmaceutically acceptable excipients.
  • the present application also provides articles of manufacture comprising any one of the antibody compositions or kits described herein.
  • articles of manufacture include vials (including sealed vials).
  • Example 1 Phage display library screen for antibodies that bind Fibrin
  • Antibody screening using two human phage display libraries was performed by the following protocol.
  • phage panning was performed, followed by phage enzyme-linked immunosorbent assays (ELISA) screening.
  • ELISA phage enzyme-linked immunosorbent assays
  • IgG reformatting and antibody production was performed followed by IgG charcterization by ELISA.
  • Each library had three panning arms and 5 rounds of selection as shown in Figure 1.
  • four reagents were used: Fibrin (thrombin cleaved fibrinogen), FGG (globular domain of Fibrinogen Gamma Chain), P2 peptide of FGG, and P13 peptide of Fibrinogen Beta chain (FGB).
  • IgG reformatted antibody clones obtained from the phage display screen as described in Example 1 were further characterized by ELISA for binding to Fibrin, FGG, P2 peptide, and Fibrinogen ( Figures 4-10). Reformatted clones obtained from either the hyperimmune phage display antibody or structural scFv phage library displayed strong binding to fibrin and negligible binding to fibrinogen.
  • a selected clone (TB241-03) was further examined for binding to recombinant biotinylated FGG having the F168A/F303A mutations ( Figure 11).
  • the protocol for expression and purification of FGG-F168A/F303A mutant was as performed as follows: On Day 0 - Cell preparation and transient transfection was performed. 293-BalanCD cells were seeded at 2.9 x 106 cells/ml in 293-BalanCD media in Erlenmeyer flasks. Incubator was set at: 6% CO2, 120 rpm, 37 deg. Transfection preparation was then preformed. The amounts were based on a 0.7 L transfection volume. 1 mg/ml cDNA concentration was used in 35 ml of OptiMEM media. 0.5 mg light chain and 0.5 mg heavy chain plasmid DNA was used. OptiMEM was transferred to two 50 ml Falcon tubes.
  • His tagged FGG protein was eluted with 5 CV of Elution buffer (20 mM Tris, 150 mM NaCl, 250 mM Imidazole, pH 8). The eluate was then dialyzed into lx PBS for storage at -80 deg. and protein analysis.
  • FIG. 11 depicts the results of selected clones binding to recombinant biotinylated - FGG F168A/F303A, demonstrating that clone TB239-02 binds strongly to the recombinant mutant FGG
  • Example 3 Anti-Fibrin antibody delays and reduces clinical symptoms of disease in rodent EAE model of Multiple Sclerosis
  • mice were induced with EAE disease by injection of PLP131-151 peptide 2 hours post-injection, mice were immunized with a single injection of 60 ng/doseof Pertussis toxin.
  • Antibody clone TB239-02 was dosed at 15 mg/Kg on days 0 and 1 after immunization.
  • the purified antibody clones described herein are formulated into a pharmaceutical composition to be administered to patients for the treatment of a neurodegenerative disease (e.g., multiple sclerosis or Alzheimer’s Disease).
  • a neurodegenerative disease e.g., multiple sclerosis or Alzheimer’s Disease.
  • the pharmaceutical composition comprising an antibody clonedescribed herein is administered at a dose sufficient to effectively reduce the symptoms of the neurodegenerative disease.
  • the pharmaceutical composition is well tolerated and does not induce significant harmful adverse effects in the patient.
  • the antibody clones described herein are investigated the ability to treat colitis in a mouse model of colitis.
  • DSS dextran sodium sulfate
  • 8- 10 week old female C57BL/6 mice are acclimated to the vivarium for at least 4 days, weighed, and randomized into treatment groups based on body weight. 2 types of studies are conducted: acute (7 days) and chronic (28 days).
  • the acute DSS study is performed by adding 2.5% DSS into drinking water for 7 days. Antibodies are administered IP every 2 days (Q2D) at 10 and 30mg/kg. Mice are euthanized on day 7 with isoflurane anesthesia, exsanguination, followed by cervical dislocation. The colon is removed and analyzed for histopathology.
  • the chronic DSS study is performed by adding 2.0% DSS into drinking water for 1 week followed by replacing with 1 week of normal drinking water, followed by another week of 2% DSS and ending with another week of normal drinking water.
  • Antibody clones described herein are administered IV prophylactically starting on day 0 twice a week at 30 and 5 mg/kg. Mice are euthanized after 28 days with isoflurane anesthesia, exsanguination, followed by cervical dislocation. The colon is removed and analyzed for histopathology.

Abstract

Described herein are novel and improved antibodies that bind human fibrin methods of use thereof. In certain aspects, described herein are methods of inhibiting microglial activation. In certain aspects, described herein are pharmaceutical compositions comprising the antibodies that bind fibrin. In certain aspects, the antibodies and methods described herein are used for treatment of degenerative neuronal disorders.

Description

ANTIBODIES WHICH BIND HUMAN FIBRIN AND METHODS OF USE
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional Application No. 63/140,138 filed January 21, 2021, which is hereby incorporated in its entirety by reference.
SEQUENCE LISTING
[0002] Not applicable.
BACKGROUND
[0003] Degenerative neuronal disorders such as multiple sclerosis (MS) can involve inflammatory demyelination and autoimmune responses. Microglia, in particular perivascular microglia, are believed to be necessary not only for the maintenance, but also for the onset of inflammatory demyelination in central nervous system (CNS) autoimmune disease. Activation of microglia contributes to both neuronal and oligodendrocyte death via release of cytokines and nitric oxide. In MS, inflammatory processes are associated with destruction of myelin sheaths, and can also involve axonal damage that can lead to permanent functional deficits, such as paralysis and loss of vision. Resident microglia are responsible for demyelination, via their ability to phagocytose myelin and secrete proinflammatory cytokines.
[0004] In MS lesions, perivascular activation of microglia colocalizes with areas of blood brain barrier (BBB) disruption, and in vivo imaging studies have shown that BBB disruption provokes the immediate and focal activation of microglia. One of the earliest events coupled to BBB disruption in MS is leakage of the blood protein fibrinogen in the nervous system that results in perivascular deposition of fibrin. Fibrinogen is not present in the healthy CNS, but only leaks in the brain after BBB disruption, thus serving as an environmental "danger" signal. Upon conversion of fibrinogen to fibrin, the CD1 lb/CD18 integrin receptor (also referred to as: Mac-1, aMfl 2, Complement Receptor 3) binds to the fibrin and induces microglial activation leading to inflammatory demyelination. CD1 lb is the alpha chain of the receptor that regulates phagocytosis of myelin during inflammatory demyelination.
Immobilized fibrinogen and insoluble fibrin, but not soluble fibrinogen, have been identified as physiological, high-affinity ligands for Mac-1. [0005] The y377-395 epitope of the fibrin or fibrinogen yC domain is the binding epitope of fibrin to CDIIb. The fibrin y377-395 peptide functions as an inhibitor of microglia activation by blocking fibrin binding to Mac-1. Because fibrin mediates blood coagulation by binding via a distinct epitope to the platelet integrin allbp3 receptor, therapeutic agents (including antibodies), that block CD1 lb binding epitope to fibrin can reduce the damaging effects of fibrin in the nervous system without affecting its beneficial effects in blood coagulation. Therefore, safe, effective antibodies that inhibit fibrin induced microglial activation without affecting its beneficial effects in blood coagulation are needed as therapeutics for degenerative neuronal disorders that involve inflammatory demyelination.
SUMMARY
[0006] In certain aspects, described herein are isolated antibody that binds human fibrin, comprising a heavy chain comprising a variable heavy (VH) chain sequence comprising three heavy chain CDR sequences, CDR-H1, CDR-H2, and CDR-H3, and a light chain comprising a variable light (VL) chain sequence comprising three light chain CDR sequences, CDR-L1, CDR-L2, and CDR-L3, wherein: a. CDR-H1 comprises the sequence set forth in SEQ ID NO: 1, 13, 25, 37, 49, 61, 73,
85, 97, 109, 121, 133, 145, 157, 169, 181, 193, 205, 217, 229, 244, 256, 268, 280,
292, 304, 316, 328, 340, 352, 364, 376, 388, 400, 412, 424, 436, 448, 460, 472, 484,
496, 508, 520, 532, 546, or 556; b. CDR-H2 comprises the sequence set forth in SEQ ID NO: 2, 14, 26, 38, 50, 62, 74,
86, 98, 110, 122, 134, 146, 158, 170, 182, 194, 206, 218, 230, 245, 257, 269, 281,
293, 305, 317, 329, 341, 353, 365, 378, 389, 401, 413, 425, 437, 449, 461, 473, 485,
497, 509, 521, 533, 547, or 557; c. CDR-H3 comprises the sequence set forth in SEQ ID NO: 3, 15, 27, 39, 51, 63, 75,
87, 99, 111, 123, 135, 147, 159, 171, 183, 195, 207, 219, or 231, 246, 258, 270, 282,
294, 306, 318, 330, 342, 354, 366, 379, 390, 402, 414, 426, 438, 450, 462, 474, 486,
498, 510, 522, 534, 548, or 558; d. CDR-L1 comprises the sequence set forth in SEQ ID NO: 4, 16, 28, 40, 52, 64, 76,
88, 100, 112, 124, 136, 148, 160, 172, 184, 196, 208, 220, 232, 247, 259, 271, 283,
295, 307, 319, 331, 343, 355, 367, 380, 391, 403, 415, 427, 439, 451, 463, 475, 487,
499, 511, 523, 535, 549, or 559; e. CDR-L2 comprises the sequence set forth in SEQ ID NO: 5, 17, 29, 41, 53, 65, 77,
89, 101, 113, 125, 137, 149, 161, 173, 185, 197, 209, 221, 233, 248, 260, 272, 284,
296, 308, 320, 332, 344, 356, 368, 381, 392, 404, 416, 428, 440, 452, 464, 476, 488,
500, 512, 524, 536, 550, or 560; and f. CDR-L3 comprises the sequence set forth in SEQ ID NO: 6, 18, 30, 42, 54, 66, 78,
90, 102, 114, 126, 138, 150, 162, 174, 186, 198, 210, 222, 234, 249, 261, 273, 285,
297, 309, 321, 333, 345, 357, 369, 382, 393, 405, 417, 429, 441, 453, 465, 477, 489,
501, 513, 525, 537, 551, or 561.
[0007] In certain embodiments, the antibody comprises a VH sequence selected from a sequence set forth in one of SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562.
[0008] In certain embodiments, the antibody comprises a VL sequence selected from a sequence set forth in SEQ ID NO 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565.
[0009] In certain embodiments, the antibody comprises a VH sequence selected from a sequence set forth in one of SEQ ID Nos: 7, and the VL sequence set for in SEQ ID NO: 10. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 19 and a VL sequence set forth in SEQ ID NO: 22. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 31 and a VL sequence set forth in SEQ ID NO: 34. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 43 and a VL sequence set forth in SEQ ID NO: 46. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 55 and a VL sequence set forth in SEQ ID NO: 58. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 67 and a VL sequence set forth in SEQ ID NO: 70. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 67 and a VL sequence set forth in SEQ ID NO: 70. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 79 and a VL sequence set forth in SEQ ID NO: 82. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 79 and a VL sequence set forth in SEQ ID NO: 82. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 91 and a VL sequence set forth in SEQ ID NO: 94. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 103 and a VL sequence set forth in SEQ ID NO: 106. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 115 and a VL sequence set forth in SEQ ID NO: 118. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 127 and a VL sequence set forth in SEQ ID NO: 130. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 139 and a VL sequence set forth in SEQ ID NO: 142. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 151 and a VL sequence set forth in SEQ ID NO: 154. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 163 and a VL sequence set forth in SEQ ID NO: 166. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 175 and a VL sequence set forth in SEQ ID NO: 178. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 187 and a VL sequence set forth in SEQ ID NO: 190. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 199 and a VL sequence set forth in SEQ ID NO: 202. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 211 and a VL sequence set forth in SEQ ID NO: 214. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 223 and a VL sequence set forth in SEQ ID NO: 226. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 235 and a VL sequence set forth in SEQ ID NO: 238. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 250, and a VL sequence set forth in SEQ ID NO: 253. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 262 and a VL sequence set forth in SEQ ID NO: 265. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 274 and a VL sequence set forth in SEQ ID NO: 277. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 286 and a VL sequence set forth in SEQ ID NO: 289. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 298 and a VL sequence set forth in SEQ ID NO: 301. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 310, and a VL sequence set forth in SEQ ID NO: 313. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 322 and a VL sequence set forth in SEQ ID NO: 325. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 334 and a VL sequence set forth in SEQ ID NO: 337. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 346 and a VL sequence set forth in SEQ ID NO: 349. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 358 and a VL sequence set forth in SEQ ID NO: 361. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 370 and a VL sequence set forth in SEQ ID NO: 373. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 382 and a VL sequence set forth in SEQ ID NO: 385. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 394 and a VL sequence set forth in SEQ ID NO: 397. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 406 and a VL sequence set forth in SEQ ID NO: 409. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 418 and a VL sequence set forth in SEQ ID NO: 421. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 430 and a VL sequence set forth in SEQ ID NO: 433. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 442 and a VL sequence set forth in SEQ ID NO: 445. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 454 and a VL sequence set forth in SEQ ID NO: 457. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 466 and a VL sequence set forth in SEQ ID NO: 469. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 478 and a VL sequence set forth in SEQ ID NO: 481. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 490 and a VL sequence set forth in SEQ ID NO: 493. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 502 and a VL sequence set forth in SEQ ID NO: 505. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 514 and a VL sequence set forth in SEQ ID NO: 517. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 526 and a VL sequence set forth in SEQ ID NO: 529. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 538 and a VL sequence set forth in SEQ ID NO: 541. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 550 and a VL sequence set forth in SEQ ID NO: 553. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 562 and a VL sequence set forth in SEQ ID NO: 565.
[0010] In certain embodiments, the antibody is a humanized, human or chimeric antibody. In certain embodiments, the antibody is a humanized antibody. In certain embodiments, the antibody comprises a heavy chain human constant region of a class selected from IgG, IgA, IgD, IgE, and IgM. In certain embodiments, the human Fc region comprises a human heavy chain constant region of the class IgG and a subclass selected from IgGl, IgG2, IgG3, and IgG4. In certain embodiments, the human Fc region comprises wild-type, human IgGl Fc.
[0011] In certain embodiments, the human Fc domain comprises a sequence set forth in SEQ ID NO: 8, 20, 32, 44, 56, 68, 80, 92, 104, 116, 128, 140, 152, 164, 176, 188, 200, 212, 224, 236, 251, 263, 275, 287, 299, 311, 323, 335, 347, 359, 371, 383, 395, 407, 419, 431, 443, 455, 467, 479, 491, 503, 515, 527, 539, 551, or 563. In certain embodiments, the heavy chain comprises a constant heavy chain sequence set forth by SEQ ID NO: 8, 20, 32, 44, 56, 68, 80, 92, 104, 116, 128, 140, 152, 164, 176, 188, 200, 212, 224, 236, 251, 263, 275, 287, 299, 311, 323, 335, 347, 359, 371, 383, 395, 407, 419, 431, 443, 455, 467, 479, 491, 503, 515, 527, 539, 551, or 563. In certain embodiments, the light chain comprises a constant light chain sequence set forth by SEQ ID NO: 9, 21, 33, 45, 57, 69, 81, 93, 105, 117, 129, 141, 153, 165, 177, 189, 201, 213, 225, 237, 252, 264, 276, 288, 300, 312, 324, 336, 348, 360, 372, 384, 396, 408, 420, 432, 444, 456, 468, 480, 492, 504, 516, 528, 540, 552, or 564.
1. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 7, and the VL sequence set forth in SEQ ID NO: 10; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 19, and the VL sequence set forth in SEQ ID NO: 22; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 31, and the VL sequence set forth in SEQ ID NO: 34; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 43, and the VL sequence set forth in SEQ ID NO: 46; and the human Fc region comprises wildtype, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 55, and the VL sequence set forth in SEQ ID NO: 58; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 67, and the VL sequence set forth in SEQ ID NO: 70; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 79, and the VL sequence set forth in SEQ ID NO: 82; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 91, and the VL sequence set forth in SEQ ID NO: 94; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 103, and the VL sequence set forth in SEQ ID NO: 106; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 115, and the VL sequence set forth in SEQ ID NO: 118; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 127, and the VL sequence set forth in SEQ ID NO: 130; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 139, and the VL sequence set forth in SEQ ID NO: 142; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 151, and the VL sequence set forth in SEQ ID NO: 154; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 163, and the VL sequence set forth in SEQ ID NO: 166; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 175, and the VL sequence set forth in SEQ ID NO: 178; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 187, and the VL sequence set forth in SEQ ID NO: 190; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 199, and the VL sequence set forth in SEQ ID NO: 202; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 211, and the VL sequence set forth in SEQ ID NO: 214; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 223, and the VL sequence set forth in SEQ ID NO: 226; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises the VH sequence set forth in SEQ ID NO: 235, and the VL sequence set forth in SEQ ID NO: 238; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 250, and a VL sequence set forth in SEQ ID NO: 253; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 262 and a VL sequence set forth in SEQ ID NO: 265; and the human Fc region comprises wildtype, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 274 and a VL sequence set forth in SEQ ID NO: 277; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 286 and a VL sequence set forth in SEQ ID NO: 289; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 298 and a VL sequence set forth in SEQ ID NO: 301; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 310, and a VL sequence set forth in SEQ ID NO: 313; and the human Fc region comprises wildtype, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 322 and a VL sequence set forth in SEQ ID NO: 325; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 334 and a VL sequence set forth in SEQ ID NO: 337; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 346 and a VL sequence set forth in SEQ ID NO: 349; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 358 and a VL sequence set forth in SEQ ID NO: 361; and the human Fc region comprises wildtype, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 370 and a VL sequence set forth in SEQ ID NO: 373; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 382 and a VL sequence set forth in SEQ ID NO: 385; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 394 and a VL sequence set forth in SEQ ID NO: 397; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 406 and a VL sequence set forth in SEQ ID NO: 409; and the human Fc region comprises wildtype, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 418 and a VL sequence set forth in SEQ ID NO: 421; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 430 and a VL sequence set forth in SEQ ID NO: 433; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 442 and a VL sequence set forth in SEQ ID NO: 445; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 454 and a VL sequence set forth in SEQ ID NO: 457; and the human Fc region comprises wildtype, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 466 and a VL sequence set forth in SEQ ID NO: 469; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 478 and a VL sequence set forth in SEQ ID NO: 481; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 490 and a VL sequence set forth in SEQ ID NO: 493; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 502 and a VL sequence set forth in SEQ ID NO: 505; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 514 and a VL sequence set forth in SEQ ID NO: 517; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 526 and a VL sequence set forth in SEQ ID NO: 529; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 538 and a VL sequence set forth in SEQ ID NO: 541; and the human Fc region comprises wildtype, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 550 and a VL sequence set forth in SEQ ID NO: 553; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the antibody comprises a VH sequence set forth in SEQ ID NO: 562 and a VL sequence set forth in SEQ ID NO: 565; and the human Fc region comprises wild-type, human IgGl Fc. In certain embodiments, the Fc region comprises one or more amino acid substitutions, wherein the one or more substitutions result in increased antibody half-life, increased ADCC activity, increased ADCP activity, or increased CDC activity compared with the Fc without the one or more substitutions, the Fc region binds an Fey Receptor selected from the group consisting of: FcyRI, FcyRIIa, FcyRIIb, FcyRIIc, FcyRIIIa, and FcyRIIIb. the Fc region binds an Fey Receptor selected from the group consisting of: FcyRI, FcyRIIa, FcyRIIb, FcyRIIc, FcyRIIIa, and FcyRIIIb. In certain embodiments, the Fc region has reduced or is lacking Fc effector function compared to a wild-type Fc region.
[0012] In certain embodiments, the antibody binds Fibrin with a KD of less than or equal to about 1 xlO'6 M, as measured by enzyme-linked immunosorbent assay (ELISA). [0013] In certain embodiments, the antibody is a monoclonal antibody.
[0014] In certain embodiments, the antibody binds Fibrin Gamma Globular (FGG) domain. In certain embodiments, the antibody binds FGG with a KD of less than or equal to about 1 xl0-8 M, as measured by ELISA.
[0015] In certain embodiments, the antibody binds to a peptide comprising SEQ ID NO: 241 with a KD of less than or equal to about 1 xl0-8 M, as measured by ELISA.
[0016] In certain embodiments, the antibody binds to fibrinogen with a KD of greater than or equal to about 1 xl0-8 M, as measured by ELISA. In certain embodiments, the antibody inhibits Mac-1 binding to fibrin or fibrinogen yC domain. In certain embodiments, the antibody exhibits inhibition of microglial adhesion to the fibrin or fibrinogen yC domain.
[0017] In certain embodiments, the antibody exhibits activation of CD1 IB positive monocytic cells.
[0018] In certain aspects, the antibodies described herein are used for the treatment of a degenerative disorder of the nervous system.
[0019] In certain aspects, described herein is an isolated polynucleotide or set of polynucleotides encoding the antibody of any of the above claims, a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen-binding portion thereof, and optionally, wherein the polynucleotide or set of polynucleotides comprises cDNA.
[0020] In certain aspects, described herein is an isolated polynucleotide or set of polynucleotides encoding the antibody of any of the above claims, a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen-binding portion thereof, and optionally, wherein the polynucleotide or set of polynucleotides comprises cDNA.
[0021] In certain aspects, described herein is a vector or set of vectors comprising the polynucleotide or set of polynucleotides encoding the antibody of any of the above claims, a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen-binding portion thereof, and optionally, wherein the polynucleotide or set of polynucleotides comprises cDNA.
[0022] In certain aspects, described herein is a host cell comprising the polynucleotide or set of polynucleotides, or a vector or set of vectors, encoding the antibody of any of the above claims, a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen- binding portion thereof, and optionally, wherein the polynucleotide or set of polynucleotides comprises cDNA. In certain aspects, described herein is a method of producing an antibody, the method comprising expressing the antibody with the host cell and isolating the expressed antibody.
[0023] In certain aspects, described herein is a pharmaceutical composition comprising an antibody described herein and a pharmaceutically acceptable excipient.
[0024] In certain aspects, described herein is a kit comprising the antibody described herein and instructions for use.
[0025] In certain aspects, described herein is a method for treating a degenerative disorder of the nervous system, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein. In certain embodiments, the degenerative disorder of the nervous system is selected from the group consisting of: multiple sclerosis, spinal cord injury, stroke, and Alzheimer’s Disease.
[0026] In certain aspects, described herein is a method of treating a pathology associated with Mac-1 binding to fibrin or Mac-1 binding with fibrinogen, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein.
[0027] In certain aspects, described herein is a method of inhibiting microglia activation, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein.
[0028] In certain aspects, described herein is a method of preventing a degenerative disorder of the nervous system, the method comprising administering to a mammalian subject a therapeutically effective amount of the antibody or pharmaceutical composition described herein.
[0029] In certain aspects, described herein is a method of treating colitis in a subject in need thereof, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein.
[0030] In certain aspects, described herein is a method of preventing colitis in a subject in need thereof, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody or pharmaceutical composition described herein. BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS
[0031] These and other features, aspects, and advantages of the present invention will become better understood with regard to the following description, and accompanying drawings, where:
[0032] Figure l is a diagram illustrating the protocol used for screening antibodies using two phage libraries and three panning arms with five rounds of selection.
[0033] Figure 2 depicts a table and graphs showing results of phage display screen. The table (top) shows increase in binding to FGG (Fibrinogen Gamma Globular domain) after each round of selection of an exemplary clone. The three graphs depict binding of 33 exemplary clones by ELISA to FGG, Fibrinogen, and the ratio of FGG/Fibrinogen binding (bottom).
[0034] Figure 3 is a table depicting the results of the phage display screens from either the hyperimmune Fab phage library or the structural scFv phage library, showing the number of unique clones obtained wih a ratio of FGG/Fibrinogen binding greater than 2.
[0035] Figure 4 are graphs depicting binding of selected clones to P2 peptide (top-left), FGG (top-right), Fibrin (bottom-left), and Fibrinogen (bottom-right).
[0036] Figure 5 are graphs depicting binding of selected clones to Fibrin (top), and FGG (bottom).
[0037] Figure 6 are graphs depicting binding of selected clones to Fibrin (top-left), FGG (top-right), and P2 peptide (bottom).
[0038] Figure 7 are graphs depicting binding of selected clones to Fibrin (top), and FGG (bottom).
[0039] Figure 8 are graphs depicting binding of selected clones to Fibrin (top-left), FGG (top-right), and P2 peptide (bottom).
[0040] Figure 9 are graphs depicting binding of selected clones to Fibrin (top-left), FGG (top-right), and P2 peptide (bottom).
[0041] Figure 10 are graphs depicting binding of selected clones to Fibrin (top), and FGG (bottom). [0042] Figure 11 is a graph depicting binding of select clones to recombinant fibrinogen gamma chain (biotinylated- histidine tagged-FGG-F168A/F303A).
[0043] Figure 12 is a graph depicting reduction of clinical score of a rodent experimental autoimmune encephalomyelitis (EAE) model of multiple sclerosis in mice treated with 15 mg per kg of the indicated antibody clone.
DETAILED DESCRIPTION
Definitions
[0044] Unless otherwise defined, all terms of art, notations and other scientific terminology used herein are intended to have the meanings commonly understood by those of skill in the art. In some cases, terms with commonly understood meanings are defined herein for clarity and/or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a difference over what is generally understood in the art. The techniques and procedures described or referenced herein are generally well understood and commonly employed using conventional methodologies by those skilled in the art, such as, for example, the widely utilized molecular cloning methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual 4th ed. (2012) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. As appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer-defined protocols and conditions unless otherwise noted.
[0045] As used herein, the singular form “a”, “an”, and “the” includes plural references unless indicated otherwise.
[0046] It is understood that aspects and embodiments of the invention described herein include “comprising,” “consisting,” and “consisting essentially of’ aspects and embodiments. [0047] For all compositions described herein, and all methods using a composition described herein, the compositions can either comprise the listed components or steps, or can “consist essentially of’ the listed components or steps. When a composition is described as “consisting essentially of’ the listed components, the composition contains the components listed, and may contain other components which do not substantially affect the condition being treated, but do not contain any other components which substantially affect the condition being treated other than those components expressly listed; or, if the composition does contain extra components other than those listed which substantially affect the condition being treated, the composition does not contain a sufficient concentration or amount of the extra components to substantially affect the condition being treated. When a method is described as “consisting essentially of’ the listed steps, the method contains the steps listed, and may contain other steps that do not substantially affect the condition being treated, but the method does not contain any other steps which substantially affect the condition being treated other than those steps expressly listed. As a non-limiting specific example, when a composition is described as ‘consisting essentially of a component, the composition may additionally contain any amount of pharmaceutically acceptable carriers, vehicles, or diluents and other such components which do not substantially affect the condition being treated. [0048] The term “vector,” as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes the vector as a selfreplicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as “expression vectors.”
[0049] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells into which an exogenous nucleic acid has been introduced, and the progeny of such cells. Host cells include “transformants” (or “transformed cells”) and “transfectants” (or “transfected cells”), which each include the primary transformed or transfected cell and progeny derived therefrom. Such progeny may not be completely identical in nucleic acid content to a parent cell, and may contain mutations. A “recombinant host cell” or “host cell” refers to a cell that includes an exogenous polynucleotide, regardless of the method used for insertion, for example, direct uptake, transduction, f-mating, or other methods known in the art to create recombinant host cells.
[0050] As used herein, the term “eukaryote” refers to organisms belonging to the phylogenetic domain Eucarya such as animals (including but not limited to, mammals, insects, reptiles, birds, etc.), ciliates, plants (including but not limited to, monocots, dicots, algae, etc.), fungi, yeasts, flagellates, microsporidia, protists, etc.
[0051] As used herein, the term “prokaryote” refers to prokaryotic organisms. For example, a non-eukaryotic organism can belong to the Eubacteria (including but not limited to, Escherichia coli, Thermus thermophilus, Bacillus stearothermophilus, Pseudomonas fluorescens, Pseudomonas aeruginosa, Pseudomonas putida, etc.) phylogenetic domain, or the Archaea (including but not limited to, Methanococcus jannaschii, Methanobacterium thermoautotrophicum, Halobacterium such as Haloferax volcanii and Halobacterium species NRC-1, Archaeoglobus fulgidus, Pyrococcus furiosus, Pyrococcus horikoshii, Aeuropyrum pernix, etc.) phylogenetic domain.
[0052] An “effective amount” or “therapeutically effective amount” as used herein refers to an amount of therapeutic compound, such as an anti-FIBRIN antibody, administered to an individual, either as a single dose or as part of a series of doses, which is effective to produce or contribute to a desired therapeutic effect, either alone or in combination with another therapeutic modality. Examples of a desired therapeutic effect is enhancing an immune response, slowing or delaying tumor development; stabilization of disease; amelioration of one or more symptoms. An effective amount may be given in one or more dosages.
[0053] The term “treating” (and variations thereof such as “treat” or “treatment”) refers to clinical intervention in an attempt to alter the natural course of a disease or condition in a subject in need thereof. Treatment can be performed during the course of clinical pathology. Desirable effects of treatment include preventing recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.
[0054] The term “sufficient amount” means an amount sufficient to produce a desired effect, e.g., an amount sufficient to modulate an immune response in a subject.
[0055] As used herein, the term “subject” or “individual” means a mammalian subject. Exemplary subjects include humans, monkeys, dogs, cats, mice, rats, cows, horses, camels, goats, rabbits, and sheep. In certain embodiments, the subject is a human. In some embodiments the subject has a disease or condition that can be treated with an antibody provided herein. In some aspects, the disease or condition is a cancer. In some aspects, the disease or condition is a viral infection.
[0056] The term “in vitro” refers to processes that occur in a living cell growing separate from a living organism, e.g., growing in tissue culture.
[0057] The term “in vivo” refers to processes that occur in a living organism.
[0058] The term “package insert” is used to refer to instructions customarily included in commercial packages of therapeutic or diagnostic products (e.g., kits) that contain information about the indications, usage, dosage, administration, combination therapy, contraindications and/or warnings concerning the use of such therapeutic or diagnostic products. [0059] The term “pharmaceutical composition” refers to a preparation which is in such form as to permit the biological activity of an active ingredient contained therein to be effective in treating a subject, and which contains no additional components which are unacceptably toxic to the subject in the amounts provided in the pharmaceutical composition.
[0060] The terms “co-administration”, “co-administer”, and “in combination with” include the administration of two or more therapeutic agents either simultaneously, concurrently or sequentially within no specific time limits. In one embodiment, the agents are present in the cell or in the subject's body at the same time or exert their biological or therapeutic effect at the same time. In one embodiment, the therapeutic agents are in the same composition or unit dosage form. In other embodiments, the therapeutic agents are in separate compositions or unit dosage forms. In certain embodiments, a first agent can be administered prior to the administration of a second therapeutic agent.
[0061] The terms “modulate” and “modulation” refer to reducing or inhibiting or, alternatively, activating or increasing, a recited variable.
[0062] The terms “increase” and “activate” refer to an increase of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or greater in a recited variable.
[0063] The terms “reduce” and “inhibit” refer to a decrease of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50- fold, 100-fold, or greater in a recited variable.
[0064] The term “about” indicates and encompasses an indicated value and a range above and below that value. In certain embodiments, the term “about” indicates the designated value ± 10%, ± 5%, or ± 1%. In certain embodiments, where applicable, the term “about” indicates the designated value(s) ± one standard deviation of that value(s).
[0065] The term “agonize” refers to the activation of receptor signaling to induce a biological response associated with activation of the receptor. An “agonist” is an entity that binds to and agonizes a receptor.
[0066] The term “antagonize” refers to the inhibition of receptor signaling to inhibit a biological response associated with activation of the receptor. An “antagonist” is an entity that binds to and antagonizes a receptor. [0067] For any of the structural and functional characteristics described herein, methods of determining these characteristics are known in the art.
[0068] The term “optionally” is meant, when used sequentially, to include from one to all of the enumerated combinations and contemplates all sub-combinations.
[0069] The term “amino acid” refers to the twenty common naturally occurring amino acids. Naturally occurring amino acids include alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine (Cys; C); glutamic acid (Glu; E), glutamine (Gin; Q), Glycine (Gly; G); histidine (His; H), isoleucine (He; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; S), threonine (Thr; T), tryptophan (Trp; W), tyrosine (Tyr; Y), and valine (Vai; V).
[0070] The term “affinity” refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen or epitope). Unless indicated otherwise, as used herein, “affinity” refers to intrinsic binding affinity, which reflects a 1 : 1 interaction between members of a binding pair (e.g., antibody and antigen or epitope).
[0071] The term “kd” (sec-1), as used herein, refers to the dissociation rate constant of a particular antibody - antigen interaction. This value is also referred to as the koff value.
[0072] The term “ka” (M-l *sec-l), as used herein, refers to the association rate constant of a particular antibody -antigen interaction. This value is also referred to as the kon value.
[0073] The term “KD” (M), as used herein, refers to the dissociation equilibrium constant of a particular antibody -antigen interaction. KD = kd/ka. In some embodiments, the affinity of an antibody is described in terms of the KD for an interaction between such antibody and its antigen. For clarity, as known in the art, a smaller KD value indicates a higher affinity interaction, while a larger KD value indicates a lower affinity interaction.
[0074] The term “KA” (M-l), as used herein, refers to the association equilibrium constant of a particular antibody-antigen interaction. KA = ka/kd.
[0075] The term “antibody” is used herein in its broadest sense and includes certain types of immunoglobulin molecules comprising one or more antigen-binding domains that specifically bind to an antigen or epitope. An antibody specifically includes intact antibodies (e.g., intact immunoglobulins), antibody fragments, and multi-specific antibodies. [0076] A “Fibrin antibody,” “anti-Fibrin antibody,” or “Fibrin -specific antibody” is an antibody, as provided herein, which specifically binds to the antigen Fibrin. In some embodiments, the antibody binds the extracellular domain of Fibrin. In certain embodiments, a Fibrin antibody provided herein binds to an epitope of Fibrin that is conserved between or among Fibrin proteins from different species.
[0077] The term “epitope” means a portion of an antigen that specifically binds to an antibody.
[0078] The term “hypervariable region” or “HVR”, as used herein, refers to each of the regions of an antibody variable domain which are hypervariable in sequence and/or form structurally defined loops (“hypervariable loops”).
[0079] The term “antigen-binding domain” means the portion of an antibody that is capable of specifically binding to an antigen or epitope.
[0080] The term “chimeric antibody” refers to an antibody in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.
[0081] The term “human antibody” refers to an antibody which possesses an amino acid sequence corresponding to that of an antibody produced by a human or a human cell, or derived from a non-human source that utilizes a human antibody repertoire or human antibody-encoding sequences (e.g., obtained from human sources or designed de novo). Human antibodies specifically exclude humanized antibodies.
[0082] The term “humanized antibody” refers to a protein having a sequence that differs from the sequence of an antibody derived from a non-human species by one or more amino acid substitutions, deletions, and/or additions, such that the humanized antibody is less likely to induce an immune response, and/or induces a less severe immune response, as compared to the non-human species antibody, when it is administered to a human subject.
[0083] The term “multispecific antibody” refers to an antibody that comprises two or more different antigen-binding domains that collectively specifically bind two or more different epitopes.
[0084] A “monospecific antibody” is an antibody that comprises one or more binding sites that specifically bind to a single epitope. An example of a monospecific antibody is a naturally occurring IgG molecule which, while divalent (i.e., having two antigen-binding domains), recognizes the same epitope at each of the two antigen-binding domains. The binding specificity may be present in any suitable valency.
[0085] The term “monoclonal antibody” refers to an antibody from a population of substantially homogeneous antibodies. A population of substantially homogeneous antibodies comprises antibodies that are substantially similar and that bind the same epitope(s), except for variants that may normally arise during production of the monoclonal antibody. Such variants are generally present in only minor amounts. A monoclonal antibody is typically obtained by a process that includes the selection of a single antibody from a plurality of antibodies. For example, the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, yeast clones, bacterial clones, or other recombinant DNA clones. The selected antibody can be further altered, for example, to improve affinity for the target (“affinity maturation”), to humanize the antibody, to improve its production in cell culture, and/or to reduce its immunogenicity in a subject.
[0086] The term "single-chain" refers to a molecule comprising amino acid monomers linearly linked by peptide bonds. In a particular such embodiment, the C-terminus of the Fab light chain is connected to the N-terminus of the Fab heavy chain in the single-chain Fab molecule. As described in more detail herein, an scFv has a variable domain of light chain (VL) connected from its C-terminus to the N-terminal end of a variable domain of heavy chain (VH) by a polypeptide chain. Alternately the scFv comprises of polypeptide chain where in the C-terminal end of the VH is connected to the N-terminal end of VL by a polypeptide chain.
[0087] The “Fab fragment” (also referred to as fragment antigen-binding) contains the constant domain (CL) of the light chain and the first constant domain (CHI) of the heavy chain along with the variable domains VL and VH on the light and heavy chains respectively. The variable domains comprise the complementarity determining loops (CDR, also referred to as hypervariable region) that are involved in antigen-binding. Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CHI domain including one or more cysteines from the antibody hinge region.
[0088] “F(ab’)2” fragments contain two Fab’ fragments joined, near the hinge region, by disulfide bonds. F(ab’)2 fragments may be generated, for example, by recombinant methods or by pepsin digestion of an intact antibody. The F(ab’) fragments can be dissociated, for example, by treatment with B-mercaptoethanol. [0089] “Fv” fragments comprise a non-covalently-linked dimer of one heavy chain variable domain and one light chain variable domain.
[0090] “Single-chain Fv” or “sFv” or “scFv” includes the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. In one embodiment, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen-binding. For a review of scFv see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994). HER2 antibody scFv fragments are described in WO93/16185; U.S. Pat. No. 5,571,894; and U.S. Pat. No. 5,587,458.
[0091] “ scFv-Fc” fragments comprise an scFv attached to an Fc domain. For example, an Fc domain may be attached to the C-terminal of the scFv. The Fc domain may follow the VH or VL, depending on the orientation of the variable domains in the scFv (i.e., VH -VL or VL - VH ). Any suitable Fc domain known in the art or described herein may be used. In some cases, the Fc domain comprises an IgG4 Fc domain.
[0092] The term “single domain antibody” or “sdAb” refers to a molecule in which one variable domain of an antibody specifically binds to an antigen without the presence of the other variable domain. Single domain antibodies, and fragments thereof, are described in Arabi Ghahroudi et al., FEBS Letters, 1998, 414:521-526 and Muyldermans et al., Trends in Biochem. Sci., 2001, 26:230-245, each of which is incorporated by reference in its entirety. Single domain antibodies are also known as sdAbs or nanobodies. Sdabs are fairly stable and easy to express as fusion partner with the Fc chain of an antibody (Harmsen MM, De Haard HJ (2007). "Properties, production, and applications of camelid single-domain antibody fragments". Appl. Microbiol Biotechnol. 77(1): 13-22).
[0093] The terms “full length antibody,” “intact antibody,” and “whole antibody” are used herein interchangeably to refer to an antibody having a structure substantially similar to a naturally occurring antibody structure and having heavy chains that comprise an Fc region. For example, when used to refer to an IgG molecule, a “full length antibody” is an antibody that comprises two heavy chains and two light chains.
[0094] The term “antibody fragment” refers to an antibody that comprises a portion of an intact antibody, such as the antigen-binding or variable region of an intact antibody. Antibody fragments include, for example, Fv fragments, Fab fragments, F(ab’)2 fragments, Fab’ fragments, scFv (sFv) fragments, and scFv-Fc fragments.
[0095] The term “Fc domain” or “Fc region” herein is used to define a C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The term includes native sequence Fc regions and variant Fc regions.
[0096] The term “substantially purified” refers to a construct described herein, or variant thereof that may be substantially or essentially free of components that normally accompany or interact with the protein as found in its naturally occurring environment, i.e. a native cell, or host cell in the case of recombinantly produced heteromultimer that in certain embodiments, is substantially free of cellular material includes preparations of protein having less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1% (by dry weight) of contaminating protein.
[0097] The term percent “identity,” in the context of two or more nucleic acid or polypeptide sequences, refer to two or more sequences or subsequences that have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., using publicly available computer software such as BLAST, BLASTP, BLASTN, BLAST-2, ALIGN, MEGALIGN (DNASTAR), CLUSTALW, CLUSTAL OMEGA, or MUSCLE software or other algorithms available to persons of skill) or by visual inspection. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (ncbi.nlm.nih.gov). Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. Depending on the application, the percent "identity" can exist over a region of the sequence being compared, e.g., over a functional domain, or, alternatively, exist over the full length of the two sequences to be compared.
[0098] For sequence comparison, typically one sequence acts as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters.
[0099] Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see generally Ausubel et al., infra).
[00100] Ranges recited herein are understood to be shorthand for all of the values within the range, inclusive of the recited endpoints. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, and 50.
[00101] It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.
ANTI-FIBRIN ANTIBODIES
Antibody Structure
[00102] The present application provides antibodies and compositions comprising an antibody which binds a fibrin protein.
[00103] The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. The "class" of an antibody or immunoglobulin refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, 6, a, y, and p, respectively.
[00104] An exemplary immunoglobulin (antibody) structural unit is composed of two pairs of polypeptide chains, each pair having one “light” (about 25 kD) and one “heavy” chain (about 50-70 kD). The N-terminal domain of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these light and heavy chain domains respectively. The IgGl heavy chain comprises of the VH, CHI, CH2 and CH3 domains respectively from the N to C-terminus. The light chain comprises of the VL and CL domains from N to C terminus. The IgGl heavy chain comprises a hinge between the CHI and CH2 domains. In certain embodiments, the immunoglobulin constructs comprise at least one immunoglobulin domain from IgG, IgM, IgA, IgD, or IgE connected to a therapeutic polypeptide. In some embodiments, the immunoglobulin domain found in an antibody provided herein, is from or derived from an immunoglobulin based construct such as a diabody, or a nanobody. In certain embodiments, the immunoglobulin constructs described herein comprise at least one immunoglobulin domain from a heavy chain antibody such as a camelid antibody. In certain embodiments, the immunoglobulin constructs provided herein comprise at least one immunoglobulin domain from a mammalian antibody such as a bovine antibody, a human antibody, a camelid antibody, a mouse antibody or any chimeric antibody.
[00105] In some embodiments, the antibodies provided herein comprise a heavy chain. In one embodiment, the heavy chain is an IgA. In one embodiment, the heavy chain is an IgD. In one embodiment, the heavy chain is an IgE. In one embodiment, the heavy chain is an IgG. In one embodiment, the heavy chain is an IgM. In one embodiment, the heavy chain is an IgGl. In one embodiment, the heavy chain is an IgG2. In one embodiment, the heavy chain is an IgG3. In one embodiment, the heavy chain is an IgG4. In one embodiment, the heavy chain is an IgAl. In one embodiment, the heavy chain is an IgA2.
[00106] In some embodiments, an antibody is an IgGl antibody. In some embodiments, an antibody is an IgG3 antibody. In some embodiments, an antibody is an IgG2 antibody. In some embodiments, an antibody is an IgG4 antibody.
[00107] Generally, native four-chain antibodies comprise six HVRs; three in the VH (Hl, H2, H3), and three in the VL (LI, L2, L3). HVRs generally comprise amino acid residues from the hypervariable loops and/or from the complementarity determining regions (CDRs), the latter being of highest sequence variability and/or involved in antigen recognition. With the exception of CDR1 in VH, CDRs generally comprise the amino acid residues that form the hypervariable loops. Hypervariable regions (HVRs) are also referred to as “complementarity determining regions” (CDRs), and these terms are used herein interchangeably in reference to portions of the variable region that form the antigen-binding regions. This particular region has been described by Kabat et al., U.S. Dept, of Health and Human Services, Sequences of Proteins of Immunological Interest (1983) and by Chothia et al., J Mol Biol 196:901-917 (1987), where the definitions include overlapping or subsets of amino acid residues when compared against each other. Nevertheless, application of either definition to refer to a CDR of an antibody or variants thereof is intended to be within the scope of the term as defined and used herein. The exact residue numbers which encompass a particular CDR will vary depending on the sequence and size of the CDR. Those skilled in the art can routinely determine which residues comprise a particular CDR given the variable region amino acid sequence of the antibody.
[00108] The amino acid sequence boundaries of a CDR can be determined by one of skill in the art using any of a number of known numbering schemes, including those described by Kabat et al., supra (“Kabat” numbering scheme); Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 (“Chothia” numbering scheme); MacCallum et al., 1996, J. Mol. Biol. 262:732- 745 (“Contact” numbering scheme); Lefranc et al., Dev. Comp. Immunol., 2003, 27:55-77 (“IMGT” numbering scheme); and Honegge and Pliickthun, J. Mol. Biol., 2001, 309:657-70 (“AHo” numbering scheme); each of which is incorporated by reference in its entirety.
[00109] Table A provides the positions of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 as identified by the Kabat and Chothia schemes. For CDR-H1, residue numbering is provided using both the Kabat and Chothia numbering schemes.
[00110] CDRs may be assigned, for example, using antibody numbering software, such as Abnum, available at www.bioinf.org.uk/abs/abnum/, and described in Abhinandan and Martin, Immunology, 2008, 45:3832-3839, incorporated by reference in its entirety.
Table A. Residues in CDRs according to Kabat and Chothia numbering schemes.
Figure imgf000026_0001
* The C-terminus of CDR-H1, when numbered using the Kabat numbering convention, varies between H32 and H34, depending on the length of the CDR.
[00111] The “EU numbering scheme” is generally used when referring to a residue in an antibody heavy chain constant region (e.g., as reported in Kabat et al., supra). Unless stated otherwise, the EU numbering scheme is used to refer to residues in antibody heavy chain constant regions described herein.
[00112] One example of an antigen-binding domain is an antigen-binding domain formed by a VH-VL dimer of an antibody. Another example of an antigen-binding domain is an antigen-binding domain formed by diversification of certain loops from the tenth fibronectin type III domain of an Adnectin. An antigen-binding domain can include CDRs 1, 2, and 3 from a heavy chain in that order; and CDRs 1, 2, and 3 from a light chain in that order.
[00113] Epitopes frequently consist of surface-accessible amino acid residues and/or sugar side chains and may have specific three-dimensional structural characteristics, as well as specific charge characteristics. Conformational and non-conformational epitopes are distinguished in that the binding to the former but not the latter may be lost in the presence of denaturing solvents. An epitope may comprise amino acid residues that are directly involved in the binding, and other amino acid residues, which are not directly involved in the binding. The epitope to which an antibody binds can be determined using known techniques for epitope determination such as, for example, testing for antibody binding to Fibrin variants with different point-mutations, or to chimeric Fibrin variants.
[00114] To screen for antibodies which bind to an epitope on a target antigen bound by an antibody of interest (e.g., Fibrin), a routine cross-blocking assay such as that described in Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory, Ed Harlow and David Lane (1988), can be performed. Alternatively, or additionally, epitope mapping can be performed by methods known in the art.
[00115] Chimeric antibodies are antibodies in which a portion of the heavy and/or light chain is derived from a particular source or species, while the remainder of the heavy and/or light chain is derived from a different source or species.
[00116] Human antibodies are antibodies which possesses an amino acid sequence corresponding to that of an antibody produced by a human or a human cell, or derived from a non-human source that utilizes a human antibody repertoire or human antibody-encoding sequences (e.g., obtained from human sources or designed de novo). Human antibodies specifically exclude humanized antibodies.
[00117] A humanized antibody has a sequence that differs from the sequence of an antibody derived from a non-human species by one or more amino acid substitutions, deletions, and/or additions, such that the humanized antibody is less likely to induce an immune response, and/or induces a less severe immune response, as compared to the non-human species antibody, when it is administered to a human subject. In one embodiment, certain amino acids in the framework and constant domains of the heavy and/or light chains of the non- human species antibody are mutated to produce the humanized antibody. In another embodiment, the constant domain(s) from a human antibody are fused to the variable domain(s) of a non-human species. In another embodiment, one or more amino acid residues in one or more CDR sequences of a non-human antibody are changed to reduce the likely immunogenicity of the non-human antibody when it is administered to a human subject, wherein the changed amino acid residues either are not critical for immunospecific binding of the antibody to its antigen, or the changes to the amino acid sequence that are made are conservative changes, such that the binding of the humanized antibody to the antigen is not significantly worse than the binding of the non-human antibody to the antigen. Examples of how to make humanized antibodies can be found in U.S. Pat. Nos. 6,054,297, 5,886,152 and 5,877,293. For further details, see Jones et al., Nature, 1986, 321 :522-525; Riechmann et al., Nature, 1988, 332:323-329; and Presta, Curr. Op. Struct. Biol., 1992, 2:593-596, each of which is incorporated by reference in its entirety.
[00118] The two or more different epitopes may be epitopes on the same antigen (e.g., a single Fibrin molecule expressed by a cell) or on different antigens (e.g., different Fibrin molecules expressed by the same cell, or a Fibrin molecule and a non- Fibrin molecule). In some aspects, a multi-specific antibody binds two different epitopes (i.e., a “bispecific antibody”). In some aspects, a multi-specific antibody binds three different epitopes (i.e., a “trispecific antibody”).
[00119] Anti-Fibrin antibodies can include those described herein such as the clones set forth in the drawings and/or tables. In some embodiments, the antibody comprises an alternative scaffold. In some embodiments, the antibody consists of an alternative scaffold. In some embodiments, the antibody consists essentially of an alternative scaffold. In some embodiments, the antibody comprises an antibody fragment. In some embodiments, the antibody consists of an antibody fragment. In some embodiments, the antibody consists essentially of an antibody fragment.
[00120] In some embodiments the antibodies are monoclonal antibodies.
[00121] In some embodiments the antibodies are polyclonal antibodies.
[00122] In some embodiments the antibodies are produced by hybridomas. In other embodiments, the antibodies are produced by recombinant cells engineered to express the desired variable and constant domains.
[00123] In some embodiments the antibodies may be single chain antibodies or other antibody derivatives retaining the antigen specificity and the lower hinge region or a variant thereof.
[00124] In some embodiments the antibodies may be polyfunctional antibodies, recombinant antibodies, human antibodies, humanized antibodies, fragments or variants thereof. In particular embodiments, the antibody fragment or a derivative thereof is selected from a Fab fragment, a Fab '2 fragment, a CDR and ScFv.
[00125] In some embodiments, the antibodies are capable of forming an immune complex. For example, an immune complex can be a tumor cell covered by antibodies.
[00126] For sequence comparison, typically one sequence acts as a reference sequence to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters.
[00127] Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nafl. Acad. Sci. USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection (see generally Ausubel et al., infra). [00128] One example of an algorithm that is suitable for determining percent sequence identity and sequence similarity is the BLAST algorithm, which is described in Altschul et al., J. Mol. Biol. 215:403-410 (1990). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (www.ncbi.nlm.nih.gov/).
Sequences of Fibrin Antibodies VH Domains
[00129] In some embodiments, an antibody provided herein comprises a VH sequence selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562.
[00130] In some embodiments, an antibody provided herein comprises a VH sequence having at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% identity to an illustrative VH sequence provided in SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562. In some embodiments, an antibody provided herein comprises a VH sequence provided in SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562, with up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
VL Domains
[00131] In some embodiments, an antibody provided herein comprises a VL sequence selected from SEQ ID NOs: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397,
409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565.
[00132] In some embodiments, an antibody provided herein comprises a VL sequence having at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% identity to an illustrative VL sequence provided in SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373,
385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565. In some embodiments, an antibody provided herein comprises a VL sequence provided in SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226,
238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445,
457, 469, 481, 493, 505, 517, 529, 541, 553, or 565 with up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,
12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
VH-VL Combinations
[00133] In some embodiments, an antibody provided herein comprises a VH sequence selected from SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562; and a VL sequence selected from SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397,
409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565.
[00134] In certain aspects, any of SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346,
358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or
562can be combined with any of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565.
[00135] In some embodiments, an antibody provided herein comprises a VH sequence having at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% identity to an illustrative VH sequence provided in SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382,
394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562; and a VL sequence having at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% identity to an illustrative VL sequence provided in SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118,
130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337,
349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or
565. In some embodiments, an antibody provided herein comprises a VH sequence provided in SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562, with up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acid substitutions, and a VL sequence provided in SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565, with up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
CDRs
[00136] In some embodiments, an antibody provided herein comprises one to three CDRs of a VH domain selected from SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562. In some embodiments, an antibody provided herein comprises two to three CDRs of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562. In some embodiments, an antibody provided herein comprises three CDRs of a VH domain selected from SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562. In some aspects, the CDRs are Exemplary CDRs. In some aspects, the CDRs are Kabat CDRs. In some aspects, the CDRs are Chothia CDRs. In some aspects, the CDRs are AbM CDRs. In some aspects, the CDRs are Contact CDRs. In some aspects, the CDRs are IMGT CDRs.
[00137] In some embodiments, the CDRs are CDRs having at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1, CDR-H2, or CDR-H3 of SEQ ID NOs:7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490,
502, 514, 526, 538, 550, or 562. In some embodiments, the CDR-H1 is a CDR-H1 of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370,
382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562, with up to
1, 2, 3, 4, or 5 amino acid substitutions. In some embodiments, the CDR-H2 is a CDR-H2 of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, the CDR-H3 is a CDR-H3 of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
[00138] In some embodiments, an antibody provided herein comprises one to three CDRs of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385,
397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565. In some embodiments, an antibody provided herein comprises two to three CDRs of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433,
445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565. In some embodiments, an antibody provided herein comprises three CDRs of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565. In some aspects, the CDRs are Exemplary CDRs. In some aspects, the CDRs are Kabat CDRs. In some aspects, the CDRs are Chothia CDRs. In some aspects, the CDRs are AbM CDRs. In some aspects, the CDRs are Contact CDRs. In some aspects, the CDRs are IMGT CDRs.
[00139] In some embodiments, the CDRs are CDRs having at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1, CDR-L2, or CDR-L3 of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565. In some embodiments, the CDR-L1 is a CDR-L1 of a VL domain of SEQ ID NO: 21, with up to 1, 2, 3, 4, or 5 amino acid substitutions. In some embodiments, the CDR-L2 is a CDR-L2 of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some embodiments, the CDR-L3 is a CDR-L3 of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
[00140] In some embodiments, an antibody provided herein comprises one to three CDRs of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358,
370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562 and one to three CDRs of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337,
349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or
565. In some embodiments, an antibody provided herein comprises two to three CDRs of a VH domain selected from SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358,
370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562and two to three CDRs of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337,
349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or
565. In some embodiments, an antibody provided herein comprises three CDRs of a VH domain selected from SEQ ID NOs: , 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382,
394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562and three CDRs of a VL domain of SEQ ID NO: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385,
397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565. In some aspects, the CDRs are Exemplary CDRs. In some aspects, the CDRs are Kabat CDRs. In some aspects, the CDRs are Chothia CDRs. In some aspects, the CDRs are AbM CDRs. In some aspects, the CDRs are Contact CDRs. In some aspects, the CDRs are IMGT CDRs. [00141] In some embodiments, an antibody provided herein comprises a CDR-H3 selected of SEQ ID NOs: 3, 15, 27, 39, 51, 63, 75, 87, 99, 111, 123, 135, 147, 159, 171, 183, 195, 207, 219, or 231, 246, 258, 270, 282, 294, 306, 318, 330, 342, 354, 366, 379, 390, 402, 414, 426, 438, 450, 462, 474, 486, 498, 510, 522, 534, 548, or 558. In some aspects, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID
NOs: 3, 15, 27, 39, 51, 63, 75, 87, 99, 111, 123, 135, 147, 159, 171, 183, 195, 207, 219, or 231, 246, 258, 270, 282, 294, 306, 318, 330, 342, 354, 366, 379, 390, 402, 414, 426, 438,
450, 462, 474, 486, 498, 510, 522, 534, 548, or 558. In some embodiments, the CDR-H3 is a
CDR-H3 selected of SEQ ID NO:3, 15, 27, 39, 51, 63, 75, 87, 99, 111, 123, 135, 147, 159, 171, 183, 195, 207, 219, or 231, 246, 258, 270, 282, 294, 306, 318, 330, 342, 354, 366, 379, 390, 402, 414, 426, 438, 450, 462, 474, 486, 498, 510, 522, 534, 548, or 558, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
[00142] In some embodiments, an antibody provided herein comprises a CDR-H1 of SEQ ID NO: 1, 13, 25, 37, 49, 61, 73, 85, 97, 109, 121, 133, 145, 157, 169, 181, 193, 205, 217, 229, 244, 256, 268, 280, 292, 304, 316, 328, 340, 352, 364, 376, 388, 400, 412, 424, 436,
448, 460, 472, 484, 496, 508, 520, 532, 546, or 556. In some aspects, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 1, 13, 25, 37, 49, 61, 73, 85, 97, 109, 121, 133, 145, 157, 169, 181, 193, 205, 217, 229, 244,
256, 268, 280, 292, 304, 316, 328, 340, 352, 364, 376, 388, 400, 412, 424, 436, 448, 460,
472, 484, 496, 508, 520, 532, 546, or 556. In some embodiments, the CDR-H1 is a CDR-H1 of SEQ ID NO: 1, 13, 25, 37, 49, 61, 73, 85, 97, 109, 121, 133, 145, 157, 169, 181, 193, 205, 217, 229, 244, 256, 268, 280, 292, 304, 316, 328, 340, 352, 364, 376, 388, 400, 412, 424, 436, 448, 460, 472, 484, 496, 508, 520, 532, 546, or 556, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
[00143] In some embodiments, an antibody provided herein comprises a CDR-H2 selected of SEQ ID NO: 2, 14, 26, 38, 50, 62, 74, 86, 98, 110, 122, 134, 146, 158, 170, 182, 194, 206, 218, 230, 245, 257, 269, 281, 293, 305, 317, 329, 341, 353, 365, 378, 389, 401, 413, 425,
437, 449, 461, 473, 485, 497, 509, 521, 533, 547, or 557. In some aspects, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 2, 14, 26, 38, 50, 62, 74, 86, 98, 110, 122, 134, 146, 158, 170, 182, 194, 206, 218, 230, 245,
257, 269, 281, 293, 305, 317, 329, 341, 353, 365, 378, 389, 401, 413, 425, 437, 449, 461,
473, 485, 497, 509, 521, 533, 547, or 557. In some embodiments, the CDR-H2 is a CDR-H2 selected of SEQ ID NO: 2, 14, 26, 38, 50, 62, 74, 86, 98, 110, 122, 134, 146, 158, 170, 182, 194, 206, 218, 230, 245, 257, 269, 281, 293, 305, 317, 329, 341, 353, 365, 378, 389, 401, 413, 425, 437, 449, 461, 473, 485, 497, 509, 521, 533, 547, or 557, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
[00144] In some embodiments, an antibody provided herein comprises a CDR-L3 of SEQ ID NO: 6, 18, 30, 42, 54, 66, 78, 90, 102, 114, 126, 138, 150, 162, 174, 186, 198, 210, 222, 234, 249, 261, 273, 285, 297, 309, 321, 333, 345, 357, 369, 382, 393, 405, 417, 429, 441, 453,
465, 477, 489, 501, 513, 525, 537, 551, or 561. In some aspects, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 6, 18, 30, 42, 54, 66, 78, 90, 102, 114, 126, 138, 150, 162, 174, 186, 198, 210, 222, 234, 249, 261, 273, 285, 297, 309, 321, 333, 345, 357, 369, 382, 393, 405, 417, 429, 441, 453, 465, 477, 489,
501, 513, 525, 537, 551, or 561. In some embodiments, the CDR-L3 is a CDR-L3 of SEQ ID NO: 6, 18, 30, 42, 54, 66, 78, 90, 102, 114, 126, 138, 150, 162, 174, 186, 198, 210, 222, 234, 249, 261, 273, 285, 297, 309, 321, 333, 345, 357, 369, 382, 393, 405, 417, 429, 441, 453, 465, 477, 489, 501, 513, 525, 537, 551, or 561, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
[00145] In some embodiments, an antibody provided herein comprises a CDR-L2 of SEQ ID NO: 5, 17, 29, 41, 53, 65, 77, 89, 101, 113, 125, 137, 149, 161, 173, 185, 197, 209, 221, 233, 248, 260, 272, 284, 296, 308, 320, 332, 344, 356, 368, 381, 392, 404, 416, 428, 440, 452, 464, 476, 488, 500, 512, 524, 536, 550, or 560. In some aspects, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 5, 17, 29, 41, 53, 65, 77, 89, 101, 113, 125, 137, 149, 161, 173, 185, 197, 209, 221, 233, 248, 260, 272, 284, 296, 308, 320, 332, 344, 356, 368, 381, 392, 404, 416, 428, 440, 452, 464, 476, 488,
500, 512, 524, 536, 550, or 560. In some embodiments, the CDR-L2 is a CDR-L2 of SEQ ID
NO: 5, 17, 29, 41, 53, 65, 77, 89, 101, 113, 125, 137, 149, 161, 173, 185, 197, 209, 221, 233, 248, 260, 272, 284, 296, 308, 320, 332, 344, 356, 368, 381, 392, 404, 416, 428, 440, 452,
464, 476, 488, 500, 512, 524, 536, 550, or 560, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
[00146] In some embodiments, an antibody provided herein comprises a CDR-L1 of SEQ ID NO: 4, 16, 28, 40, 52, 64, 76, 88, 100, 112, 124, 136, 148, 160, 172, 184, 196, 208, 220, 232, 247, 259, 271, 283, 295, 307, 319, 331, 343, 355, 367, 380, 391, 403, 415, 427, 439, 451, 463, 475, 487, 499, 511, 523, 535, 549, or 559. In some aspects, the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 4, 16, 28, 40, 52, 64, 76, 88, 100, 112, 124, 136, 148, 160, 172, 184, 196, 208, 220, 232, 247, 259, 271, 283, 295, 307, 319, 331, 343, 355, 367, 380, 391, 403, 415, 427, 439, 451, 463, 475, 487, 499, 511, 523, 535, 549, or 559. In some embodiments, the CDR-L1 is a CDR-L1 of SEQ ID NO: 4, 16, 28, 40, 52, 64, 76, 88, 100, 112, 124, 136, 148, 160, 172, 184, 196, 208, 220, 232, 247, 259, 271, 283, 295, 307, 319, 331, 343, 355, 367, 380, 391, 403, 415, 427, 439, 451, 463, 475, 487, 499, 511, 523, 535, 549, or 559, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions. In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this paragraph are referred to herein as “variants.” In some embodiments, such variants are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
[00147] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 3, a CDR-H2 of SEQ ID NO: , a CDR-H1 of SEQ ID NO: 1, a CDR-L3 of SEQ ID NO: 6, a CDR-L2 of SEQ ID NO: 5, and a CDR-L1 of SEQ ID NO: 4. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 3, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 2, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 1, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 6, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 5, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 4. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 3, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 2, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 1, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 6, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 5, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 4, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00148] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 15, a CDR-H2 of SEQ ID NO: 14, a CDR-H1 of SEQ ID NO: 13, a CDR-L3 of SEQ ID NO: 18, a CDR-L2 of SEQ ID NO: 17, and a CDR-L1 of SEQ ID NO: 16. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 15, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 14, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 13, the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 18, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 17, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 16. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 15, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 14, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 13, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 18, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 17, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 16, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00149] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 27, a CDR-H2 of SEQ ID NO: 26, a CDR-H1 of SEQ ID NO: 25, a CDR-L3 of SEQ ID NO: 30, a CDR-L2 of SEQ ID NO: 29, and a CDR-L1 of SEQ ID NO: 28. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 27, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 26, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 25, the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 30, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 29, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 28. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 27, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 26, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 25, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 30, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 29, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 28, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00150] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 39, a CDR-H2 of SEQ ID NO: 38, a CDR-H1 of SEQ ID NO: 37, a CDR-L3 of SEQ ID NO: 42, a CDR-L2 of SEQ ID NO: 41, and a CDR-L1 of SEQ ID NO: 40. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 39, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 38, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 37, the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 42, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 41, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 40. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 39, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 38, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 37, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 42, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 41, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 40, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00151] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 51, a CDR-H2 of SEQ ID NO: 50, a CDR-H1 of SEQ ID NO: 49, a CDR-L3 of SEQ ID NO: 53, a CDR-L2 of SEQ ID NO: 52, and a CDR-L1 of SEQ ID NO: 51. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 51, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 50, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 49, the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 53, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 52, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 51. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 51, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 50, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 49, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 53, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 52, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 51, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00152] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 63, a CDR-H2 of SEQ ID NO: 62, a CDR-H1 of SEQ ID NO: 61, a CDR-L3 of SEQ ID NO: 66, a CDR-L2 of SEQ ID NO: 65, and a CDR-L1 of SEQ ID NO: 64. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 63, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 62, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 61, the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 66, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 65, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 64. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 61, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 62, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 61, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 66, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 65, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 64, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00153] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 75, a CDR-H2 of SEQ ID NO: 74, a CDR-H1 of SEQ ID NO: 73, a CDR-L3 of SEQ ID NO: 78, a CDR-L2 of SEQ ID NO: 77, and a CDR-L1 of SEQ ID NO: 76. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 75, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 74, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 73, the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 78, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 77, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 76. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 75, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 74, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 73, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 78, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 77, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 76, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00154] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 87, a CDR-H2 of SEQ ID NO: 86, a CDR-H1 of SEQ ID NO: 85, a CDR-L3 of SEQ ID NO: 90, a CDR-L2 of SEQ ID NO: 89, and a CDR-L1 of SEQ ID NO: 88. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 87, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 86, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 85, the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 90, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 89, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 88. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 87, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 86, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 85, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 90, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 89, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 88, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions. [00155] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 99, a CDR-H2 of SEQ ID NO: 98, a CDR-H1 of SEQ ID NO: 97, a CDR-L3 of SEQ ID NO: 102, a CDR-L2 of SEQ ID NO: 101, and a CDR-L1 of SEQ ID NO: 100. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 99, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 98, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 97, the CDR- L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 102, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 101, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 100. In some embodiments, the CORED is a CDR-H3 of SEQ ID NO: 99, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 98, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 97, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 102, with up to 1, 2,
3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 101, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 100, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00156] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 111, a CDR-H2 of SEQ ID NO: 110, a CDR-H1 of SEQ ID NO: 109, a CDR-L3 of SEQ ID NO: 114, a CDR-L2 of SEQ ID NO: 113, and a CDR-L1 of SEQ ID NO: 112. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 111, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 110, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 109, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 114, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 113, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 112. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 111, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 110, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 109, with up to 1, 2, 3,
4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 114, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 113, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 112, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00157] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 123, a CDR-H2 of SEQ ID NO: 122, a CDR-H1 of SEQ ID NO: 121, a CDR-L3 of SEQ ID NO: 126, a CDR-L2 of SEQ ID NO: 125, and a CDR-L1 of SEQ ID NO: 124. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 123, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 122, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 121, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 126, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 125, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 124. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 123, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 122, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 121, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 126, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 125, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 124, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00158] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 135, a CDR-H2 of SEQ ID NO: 134, a CDR-H1 of SEQ ID NO: 133, a CDR-L3 of SEQ ID NO: 138, a CDR-L2 of SEQ ID NO: 137, and a CDR-L1 of SEQ ID NO: 136. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 135, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 134, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 133, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 136, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 135, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 134. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 135, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 134, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 133, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 136, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 135, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 134, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00159] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 148, a CDR-H2 of SEQ ID NO: 147, a CDR-H1 of SEQ ID NO: 146, a CDR-L3 of SEQ ID NO: 151, a CDR-L2 of SEQ ID NO: 150, and a CDR-L1 of SEQ ID NO: 149. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 148, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 147, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 146, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 151, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 150, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 149. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 148, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 147, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 146, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 151, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 150, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 149, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00160] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 159, a CDR-H2 of SEQ ID NO: 158, a CDR-H1 of SEQ ID NO: 157, a CDR-L3 of SEQ ID NO: 162, a CDR-L2 of SEQ ID NO: 161, and a CDR-L1 of SEQ ID NO: 160. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 159, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 158, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 157, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 162, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 161, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 160. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 159, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 158, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 157, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 162, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 161, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 160, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00161] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 171, a CDR-H2 of SEQ ID NO: 170, a CDR-H1 of SEQ ID NO: 169, a CDR-L3 of SEQ ID NO: 174, a CDR-L2 of SEQ ID NO: 173, and a CDR-L1 of SEQ ID NO: 172. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 171, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 170, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 169, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 174, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 173, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 172. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 171, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 170, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 169, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 174, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 173, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 172, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00162] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 183, a CDR-H2 of SEQ ID NO: 182, a CDR-H1 of SEQ ID NO: 181, a CDR-L3 of SEQ ID NO: 186, a CDR-L2 of SEQ ID NO: 185, and a CDR-L1 of SEQ ID NO: 184. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 183, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 182, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 181, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 186, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 185, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 184. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 183, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 182, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 181, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 186, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 185, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 184, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00163] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 195, a CDR-H2 of SEQ ID NO: 194, a CDR-H1 of SEQ ID NO: 193, a CDR-L3 of SEQ ID NO: 198, a CDR-L2 of SEQ ID NO: 197, and a CDR-L1 of SEQ ID NO: 196. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 195, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 194, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 193, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 198, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 197, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 196. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 195, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 194, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 193, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 198, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 197, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 196, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions. [00164] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 207, a CDR-H2 of SEQ ID NO: 206, a CDR-H1 of SEQ ID NO: 205, a CDR-L3 of SEQ ID NO: 210, a CDR-L2 of SEQ ID NO: 209, and a CDR-L1 of SEQ ID NO: 208. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 207, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 206, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 205, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 210, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 209, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 208. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 207, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 206, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 205, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 210, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 209, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 208, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00165] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 219, a CDR-H2 of SEQ ID NO: 218, a CDR-H1 of SEQ ID NO: 217, a CDR-L3 of SEQ ID NO: 222, a CDR-L2 of SEQ ID NO: 221, and a CDR-L1 of SEQ ID NO: 220. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 219, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 218, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 217, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 222, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 221, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 220. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 219, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 218, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 217, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 222, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 221, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 220, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00166] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 231, a CDR-H2 of SEQ ID NO: 230, a CDR-H1 of SEQ ID NO: 229, a CDR-L3 of SEQ ID NO: 234, a CDR-L2 of SEQ ID NO: 233, and a CDR-L1 of SEQ ID NO: 232. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 231, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 230, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 229, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 234, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 233, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 232. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 231, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 230, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 229, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 234, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 233, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 232, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00167] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 246, a CDR-H2 of SEQ ID NO: 245, a CDR-H1 of SEQ ID NO: 244, a CDR-L3 of SEQ ID NO: 249, a CDR-L2 of SEQ ID NO: 248, and a CDR-L1 of SEQ ID NO: 247. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 246, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 245, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 244, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 249, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 248, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 247. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 246, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 245, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 244, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 249, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 248, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 247, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00168] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 258, a CDR-H2 of SEQ ID NO: 257, a CDR-H1 of SEQ ID NO: 256, a CDR-L3 of SEQ ID NO: 261, a CDR-L2 of SEQ ID NO: 260, and a CDR-L1 of SEQ ID NO: 259. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 258, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 257, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 256, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 261, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 260, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 259. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 258, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 257, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 256, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 261, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 260, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 259, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00169] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 270, a CDR-H2 of SEQ ID NO: 269, a CDR-H1 of SEQ ID NO: 268, a CDR-L3 of SEQ ID NO: 273, a CDR-L2 of SEQ ID NO: 272, and a CDR-L1 of SEQ ID NO: 271. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 270, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 269, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 268, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 273, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 272, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 271. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 270, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 269, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 268, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 273, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 272, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 271, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00170] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 282, a CDR-H2 of SEQ ID NO: 281, a CDR-H1 of SEQ ID NO: 280, a CDR-L3 of SEQ ID NO: 285, a CDR-L2 of SEQ ID NO: 284, and a CDR-L1 of SEQ ID NO: 283. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 282, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 281, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 280, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 285, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 284, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 283. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 282, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 281, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 280, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 285, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 284, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 283, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00171] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 294, a CDR-H2 of SEQ ID NO: 293, a CDR-H1 of SEQ ID NO: 292, a CDR-L3 of SEQ ID NO: 297, a CDR-L2 of SEQ ID NO: 296, and a CDR-L1 of SEQ ID NO: 295. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 294, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 293, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 292, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 297, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 296, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 295. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 294, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 293, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 292, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 297, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 296, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 295, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00172] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 306, a CDR-H2 of SEQ ID NO: 305, a CDR-H1 of SEQ ID NO: 304, a CDR-L3 of SEQ ID NO: 309, a CDR-L2 of SEQ ID NO: 308, and a CDR-L1 of SEQ ID NO: 307. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 306, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 305, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 304, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 309, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 308, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 307. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 306, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 305, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 304, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 309, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 308, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 307, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions. [00173] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 318, a CDR-H2 of SEQ ID NO: 317, a CDR-H1 of SEQ ID NO: 316, a CDR-L3 of SEQ ID NO: 321, a CDR-L2 of SEQ ID NO: 320, and a CDR-L1 of SEQ ID NO: 319. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 318, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 317, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 316, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 321, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 320, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 319. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 318, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 317, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 316, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 321, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 320, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 319, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00174] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 330, a CDR-H2 of SEQ ID NO: 329, a CDR-H1 of SEQ ID NO: 328, a CDR-L3 of SEQ ID NO: 333, a CDR-L2 of SEQ ID NO: 332, and a CDR-Ll of SEQ ID NO: 331. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 330, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 329, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 330, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 333, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 332, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 331. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 330, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 329, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 328, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 333, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 332, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 331, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00175] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 342, a CDR-H2 of SEQ ID NO: 341, a CDR-H1 of SEQ ID NO: 340, a CDR-L3 of SEQ ID NO: 345, a CDR-L2 of SEQ ID NO: 344, and a CDR-L1 of SEQ ID NO: 343. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 342, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 341, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 340, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 345, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 344, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 343. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 342, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 341, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 340, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 345, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 344, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 343, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00176] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 354, a CDR-H2 of SEQ ID NO: 353, a CDR-H1 of SEQ ID NO: 352, a CDR-L3 of SEQ ID NO: 357, a CDR-L2 of SEQ ID NO: 356, and a CDR-L1 of SEQ ID NO: 355. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 354, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 353, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 352, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 357, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 356, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 355. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 354, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 353, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 352, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 357, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 356, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 355, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00177] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 366, a CDR-H2 of SEQ ID NO: 365, a CDR-H1 of SEQ ID NO: 364, a CDR-L3 of SEQ ID NO: 369, a CDR-L2 of SEQ ID NO: 368, and a CDR-L1 of SEQ ID NO: 367. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 366, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 365, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 364, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 369, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 368, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 367. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 366, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 365, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 364, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 369, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 368, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 367, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00178] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 378, a CDR-H2 of SEQ ID NO: 377, a CDR-H1 of SEQ ID NO: 376, a CDR-L3 of SEQ ID NO: 381, a CDR-L2 of SEQ ID NO: 380, and a CDR-L1 of SEQ ID NO: 379. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 378, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 377, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 376, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 381, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 380, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 379. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 378, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 377, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 376, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 381, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 380, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 379, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00179] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 390, a CDR-H2 of SEQ ID NO: 389, a CDR-H1 of SEQ ID NO: 388, a CDR-L3 of SEQ ID NO: 393, a CDR-L2 of SEQ ID NO: 392, and a CDR-L1 of SEQ ID NO: 391. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 390, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 389, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 388, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 393, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 392, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 391. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 390, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 389, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 388, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 393, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 392, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 391, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00180] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 402, a CDR-H2 of SEQ ID NO: 401, a CDR-H1 of SEQ ID NO: 400, a CDR-L3 of SEQ ID NO: 405, a CDR-L2 of SEQ ID NO: 404, and a CDR-L1 of SEQ ID NO: 403. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 402, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 401, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 400, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 405, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 404, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 403. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 402, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 401, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 400, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 405, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 404, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 403, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00181] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 414, a CDR-H2 of SEQ ID NO: 413, a CDR-H1 of SEQ ID NO: 412, a CDR-L3 of SEQ ID NO: 417, a CDR-L2 of SEQ ID NO: 416, and a CDR-L1 of SEQ ID NO: 415. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 414, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 413, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 412, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 417, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 416, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 415. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 414, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 413, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 412, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 417, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 416, with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 415, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions. [00182] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 426, a CDR-H2 of SEQ ID NO: 425, a CDR-H1 of SEQ ID NO: 424, a CDR-L3 of SEQ ID NO: 429, a CDR-L2 of SEQ ID NO: 428, and a CDR-L1 of SEQ ID NO: 427. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 426, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 425, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 424, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 429, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 428, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 427. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 426, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 425, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 424, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 429, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 428 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 427, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00183] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 438, a CDR-H2 of SEQ ID NO: 437, a CDR-H1 of SEQ ID NO: 436, a CDR-L3 of SEQ ID NO: 441, a CDR-L2 of SEQ ID NO: 440, and a CDR-L1 of SEQ ID NO: 439. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 438, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 437, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 436, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 441, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 440, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 439. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 438, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 437, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 436, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 441, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 440 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 439, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00184] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 450, a CDR-H2 of SEQ ID NO: 449, a CDR-H1 of SEQ ID NO: 448 a CDR-L3 of SEQ ID NO: 453, a CDR-L2 of SEQ ID NO: 452, and a CDR-L1 of SEQ ID NO: 451. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 450, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 449, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 448, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 453, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 452, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 451. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 450, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 449, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 448, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 453, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 452 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 451, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00185] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 462, a CDR-H2 of SEQ ID NO: 461, a CDR-H1 of SEQ ID NO: 460 a CDR-L3 of SEQ ID NO: 465, a CDR-L2 of SEQ ID NO: 464, and a CDR-L1 of SEQ ID NO: 463. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 462, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 461, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 460, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 465, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 464, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 463. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 462, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 461, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 460, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 465, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 464 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 463, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00186] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 474, a CDR-H2 of SEQ ID NO: 473, a CDR-H1 of SEQ ID NO: 472 a CDR-L3 of SEQ ID NO: 477, a CDR-L2 of SEQ ID NO: 476, and a CDR-L1 of SEQ ID NO: 475. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 474, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 473, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 472, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 477, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 476, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 475. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 474, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 473, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 472, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 477, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 476 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 475, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00187] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 486, a CDR-H2 of SEQ ID NO: 485, a CDR-H1 of SEQ ID NO: 484 a CDR-L3 of SEQ ID NO: 489, a CDR-L2 of SEQ ID NO: 488, and a CDR-L1 of SEQ ID NO: 487. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 486, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 485, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 484, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 489, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 488, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 487. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 486, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 485, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 484, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 489, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 488 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 487, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00188] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 498, a CDR-H2 of SEQ ID NO: 497, a CDR-H1 of SEQ ID NO: 496 a CDR-L3 of SEQ ID NO: 501, a CDR-L2 of SEQ ID NO: 500, and a CDR-L1 of SEQ ID NO: 499. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 498, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 497, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 496, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 501, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 500, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 499. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 498, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 497, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 496, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 501, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 500 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 499, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00189] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 510, a CDR-H2 of SEQ ID NO: 509, a CDR-H1 of SEQ ID NO: 508 a CDR-L3 of SEQ ID NO: 513, a CDR-L2 of SEQ ID NO: 512, and a CDR-L1 of SEQ ID NO: 511. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 510, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 509, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 508, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 513, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 512, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 511. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 510, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 509, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 508, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 513, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 512 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 511, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00190] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 522, a CDR-H2 of SEQ ID NO: 521, a CDR-H1 of SEQ ID NO: 520 a CDR-L3 of SEQ ID NO: 525, a CDR-L2 of SEQ ID NO: 524, and a CDR-L1 of SEQ ID NO: 523. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 522, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 521, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 520, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 525, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 524, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 523. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 522, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 521, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 520, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 525, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 524 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 523, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions. [00191] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 534, a CDR-H2 of SEQ ID NO: 533, a CDR-H1 of SEQ ID NO: 532 a CDR-L3 of SEQ ID NO: 537, a CDR-L2 of SEQ ID NO: 536, and a CDR-L1 of SEQ ID NO: 535. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 534, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 533, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 532, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 537, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 536, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 535. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 534, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 533, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 532, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 537, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 536 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 535, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00192] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 546, a CDR-H2 of SEQ ID NO: 545, a CDR-H1 of SEQ ID NO: 544 a CDR-L3 of SEQ ID NO: 549, a CDR-L2 of SEQ ID NO: 548, and a CDR-L1 of SEQ ID NO: 547. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 546, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 545, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 544, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 549, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 548, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 547. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 546, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 545, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 544, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 549, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 548 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 547, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00193] In some embodiments, an antibody provided herein comprises a CDR-H3 of SEQ ID NO: 558, a CDR-H2 of SEQ ID NO: 557, a CDR-H1 of SEQ ID NO: 556 a CDR-L3 of SEQ ID NO: 561, a CDR-L2 of SEQ ID NO: 560, and a CDR-L1 of SEQ ID NO: 559. In some embodiments, the CDR-H3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H3 of SEQ ID NO: 558, the CDR-H2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H2 of SEQ ID NO: 557, the CDR-H1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-H1 of SEQ ID NO: 556, the CDR-L3 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L3 of SEQ ID NO: 561, the CDR-L2 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L2 of SEQ ID NO: 560, and the CDR-L1 has at least about 50%, 75%, 80%, 85%, 90%, or 95% identity with a CDR-L1 of SEQ ID NO: 559. In some embodiments, the CDR-H3 is a CDR-H3 of SEQ ID NO: 558, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H2 is a CDR-H2 of SEQ ID NO: 557, with up to 1, 2, 3, 4, 5, 6, 7, or 8 amino acid substitutions; the CDR-H1 is a CDR-H1 of SEQ ID NO: 556, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L3 is a CDR-L3 of SEQ ID NO: 561, with up to 1, 2, 3, 4, or 5 amino acid substitutions; the CDR-L2 is a CDR-L2 of SEQ ID NO: 560 with up to 1, 2, 3, or 4 amino acid substitutions; and the CDR-L1 is a CDR-L1 of SEQ ID NO: 559, with up to 1, 2, 3, 4, 5, or 6 amino acid substitutions.
[00194] In some aspects, the amino acid substitutions are conservative amino acid substitutions. In some embodiments, the antibodies described in this disclosure are referred to herein as “variants” or “clones”. In some embodiments, such variants or clones are derived from a sequence provided herein, for example, by affinity maturation, site directed mutagenesis, random mutagenesis, or any other method known in the art or described herein. In some embodiments, such variants or cones are not derived from a sequence provided herein and may, for example, be isolated de novo according to the methods provided herein for obtaining antibodies.
Fc Region
[00195] The structures of the Fc regions of various immunoglobulins, and the glycosylation sites contained therein, are known in the art. See Schroeder and Cavacini, J. Allergy Clin. Immunol., 2010, 125:S41-52, incorporated by reference in its entirety. The Fc region may be a naturally occurring Fc region, or an Fc region modified as described in the art or elsewhere in this disclosure.
[00196] Unless otherwise specified herein, numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, also called the EU index, as described in Kabat et al, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991. An "Fc polypeptide" of a dimeric Fc as used herein refers to one of the two polypeptides forming the dimeric Fc domain, i.e. a polypeptide comprising C-terminal constant regions of an immunoglobulin heavy chain, capable of stable self-association. For example, an Fc polypeptide of a dimeric IgG Fc comprises an IgG CH2 and an IgG CH3 constant domain sequence. An Fc can be of the class IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl, and IgA2.
[00197] The terms “Fc receptor” and “FcR” are used to describe a receptor that binds to the Fc region of an antibody. For example, an FcR can be a native sequence human FcR. Generally, an FcR is one which binds an IgG antibody (a gamma receptor) and includes receptors of the FcyRI, FcyRII, and FcyRIII subclasses, including allelic variants and alternatively spliced forms of these receptors. FcyRII receptors include FcyRIIA (an “activating receptor”) and FcyRIIB (an “inhibiting receptor”), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof. Immunoglobulins of other isotypes can also be bound by certain FcRs (see, e.g., Janeway et al., Immuno Biology: the immune system in health and disease, (Elsevier Science Ltd., NY) (4th ed., 1999)).
Activating receptor FcyRIIA contains an immunoreceptor tyrosine-based activation motif (IT AM) in its cytoplasmic domain. Inhibiting receptor FcyRIIB contains an immunoreceptor tyrosine-based inhibition motif (ITEM) in its cytoplasmic domain (reviewed in Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term “FcR” herein. The term also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976); and Kim et al., J. Immunol. 24:249 (1994)). [00198] Modifications in the CH2 domain can affect the binding of FcRs to the Fc. A number of amino acid modifications in the Fc region are known in the art for selectively altering the affinity of the Fc for different Fcgamma receptors. In some aspects, the Fc comprises one or more modifications to promote selective binding of Fc-gamma receptors.
[00199] Exemplary mutations that alter the binding of FcRs to the Fc are listed below:
[00200] S298A/E333A/K334A, S298A/E333A/K334A/K326A (Lu Y, Vernes JM, Chiang N, et al. J Immunol Methods. 2011 Feb 28;365(l-2): 132-41);
[00201] F243L/R292P/Y300L/V305I/P396L, F243L/R292P/Y300L/L235V/P396L (Stavenhagen JB, Gorlatov S, Tuaillon N, et al. Cancer Res. 2007 Sep 15;67(18):8882- 90; Nordstrom JL, Gorlatov S, Zhang W, et al. Breast Cancer Res. 2011 Nov 30;13(6):R123);
[00202] F243L (Stewart R, Thom G, Levens M, et al. Protein Eng Des Sei. 2011 Sep;24(9):671-8.), S298A/E333A/K334A (Shields RL, Namenuk AK, Hong K, et al. J Biol Chem. 2001 Mar 2;276(9):6591-604);
[00203] S239D/I332E/A330L, S239D/I332E (Lazar GA, Dang W, Karki S, et al. Proc Natl Acad Sci U S A. 2006 Mar 14; 103(11):4005-10);
[00204] S239D/S267E, S267E/L328F (Chu SY, Vostiar I, Karki S, et al. Mol Immunol. 2008 Sep;45(15):3926-33);
[00205] S239D/D265S/S298A/I332E, S239E/S298A/K326A/A327H, G237F/S298A/A330L /I332E, S239D/I332E/S298A, S239D/K326E/A330L/I332E/S298A, G236A/S239D/D270L/I 332E, S239E/S267E/H268D, L234F/S267E/N325L, G237F/V266L/S267D and other mutations listed in WO2011/120134 and WO2011/120135, herein incorporated by reference. Therapeutic Antibody Engineering (by William R. Strohl and Lila M. Strohl, Woodhead Publishing series in Biomedicine No 11, ISBN 1 907568 37 9, Oct 2012) lists mutations on page 283.
[00206] In some embodiments an antibody described herein includes modifications to improve its ability to mediate effector function. Such modifications are known in the art and include afucosylation, or engineering of the affinity of the Fc towards an activating receptor, mainly FCGR3a for ADCC, and towards Clq for CDC. The following Table B summarizes various designs reported in the literature for effector function engineering. [00207] Methods of producing antibodies with little or no fucose on the Fc glycosylation site (Asn 297 EU numbering) without altering the amino acid sequence are well known in the art. The GlymaX® technology (ProBioGen AG) is based on the introduction of a gene for an enzyme which deflects the cellular pathway of fucose biosynthesis into cells used for antibody production. This prevents the addition of the sugar “fucose” to the N-linked antibody carbohydrate part by antibody-producing cells, (von Horsten et al. (2010) Glycobiology. 2010 Dec; 20 (12): 1607-18. Another approach to obtaining antibodies with lowered levels of fucosylation can be found in U.S. patent 8,409,572, which teaches selecting cell lines for antibody production for their ability to yield lower levels of fucosylation on antibodies can be fully afucosylated (meaning they contain no detectable fucose) or they can be partially afucosylated, meaning that the isolated antibody contains less than 95%, less than 85%, less than 75%, less than 65%, less than 55%, less than 45%, less than 35%, less than 25%, less than 15% or less than 5% of the amount of fucose normally detected for a similar antibody produced by a mammalian expression system.
[00208] Thus, in one embodiment, an antibody described herein can include a dimeric Fc that comprises one or more amino acid modifications as noted in Table B that confer improved effector function. In another embodiment, the antibody can be afucosylated to improve effector function.
Table B: CH2 domains and effector function engineering
Figure imgf000068_0001
[00209] Fc modifications reducing FcgR and/or complement binding and/or effector function are known in the art. Recent publications describe strategies that have been used to engineer antibodies with reduced or silenced effector activity (see Strohl, WR (2009), Curr Opin Biotech 20:685-691, and Strohl, WR and Strohl LM, “Antibody Fc engineering for optimal antibody performance” In Therapeutic Antibody Engineering, Cambridge: Woodhead Publishing (2012), pp 225-249). These strategies include reduction of effector function through modification of glycosylation, use of IgG2/IgG4 scaffolds, or the introduction of mutations in the hinge or CH2 regions of the Fc. For example, US Patent Publication No. 2011/0212087 (Strohl), International Patent Publication No. WO 2006/105338 (Xencor), US Patent Publication No. 2012/0225058 (Xencor), US Patent Publication No. 2012/0251531 (Genentech), and Strop et al ((2012) J. Mol. Biol. 420: 204- 219) describe specific modifications to reduce FcgR or complement binding to the Fc.
[00210] Specific, non-limiting examples of known amino acid modifications to reduce FcgR or complement binding to the Fc include those identified in the following Table C:
Table C: Modifications to reduce FcgR or complement binding to the Fc
Figure imgf000069_0001
[00211] Methods of producing antibodies with little or no fucose on the Fc glycosylation site (Asn 297 EU numbering) without altering the amino acid sequence are well known in the art. The GlymaxX® technology (ProBioGen AG) is based on the introduction of a gene for an enzyme which deflects the cellular pathway of fucose biosynthesis into cells used for antibody production. This prevents the addition of the sugar “fucose” to the N-linked antibody carbohydrate part by antibody-producing cells, (von Horsten et al. (2010) Glycobiology. 2010 Dec; 20 (12): 1607-18.) Examples of cell lines capable of producing defucosylated antibody include CHO-DG44 with stable overexpression of the bacterial oxidoreductase GDP-6-deoxy-D-lyxo-4-hexylose reductase (RMD) (see Henning von Horsten et al., Glycobiol 2010, 20: 1607-1618) or Lecl3 CHO cells, which are deficient in protein fucosylation (see Ripka et al., Arch. Biochem. Biophys., 1986, 249:533-545; U.S. Pat. Pub. No. 2003/0157108; WO 2004/056312; each of which is incorporated by reference in its entirety), and knockout cell lines, such as alpha- 1,6-fucosyltransf erase gene or FUT8 knockout CHO cells (see Yamane-Ohnuki et al., Biotech. Bioeng., 2004, 87: 614-622; Kanda et al., Biotechnol. Bioeng., 2006, 94:680-688; and WO 2003/085107; each of which is incorporated by reference in its entirety). Another approach to obtaining antibodies with lowered levels of fucosylation can be found in U.S. patent 8,409,572, which teaches selecting cell lines for antibody production for their ability to yield lower levels of fucosylation on antibodies
[00212] Examples of cell lines capable of producing defucosylated antibody include CHO- DG44 with stable overexpression of the bacterial oxidoreductase GDP-6-deoxy-D-lyxo-4- hexylose reductase (RMD) (see Henning von Horsten et al., Glycobiol 2010, 20: 1607-1618) or Lecl3 CHO cells, which are deficient in protein fucosylation (see Ripka et al., Arch. Biochem. Biophys., 1986, 249:533-545; U.S. Pat. Pub. No. 2003/0157108; WO 2004/056312; each of which is incorporated by reference in its entirety), and knockout cell lines, such as alpha- 1,6-fucosyltransf erase gene or FUT8 knockout CHO cells (see Yamane- Ohnuki et al., Biotech. Bioeng., 2004, 87: 614-622; Kanda et al., Biotechnol. Bioeng., 2006, 94:680-688; and WO 2003/085107; each of which is incorporated by reference in its entirety).
[00213] Antibodies can be fully afucosylated (meaning they contain no detectable fucose) or they can be partially afucosylated, meaning that the isolated antibody contains less than 95%, less than 85%, less than 75%, less than 65%, less than 55%, less than 45%, less than 35%, less than 25%, less than 15% or less than 5% of the amount of fucose normally detected for a similar antibody produced by a mammalian expression system.
[00214] In some aspects, an antibody provided herein comprises an IgGl domain with reduced fucose content at position Asn 297 compared to a naturally occurring IgGl domain. Such Fc domains are known to have improved ADCC. See Shields et al., J. Biol. Chem., 2002, 277:26733-26740, incorporated by reference in its entirety. In some aspects, such antibodies do not comprise any fucose at position Asn 297. The amount of fucose may be determined using any suitable method, for example as described in WO 2008/077546, incorporated by reference in its entirety.
[00215] In certain embodiments, an antibody provided herein comprises an Fc region with one or more amino acid substitutions which improve ADCC, such as a substitution at one or more of positions 298, 333, and 334 of the Fc region. In some embodiments, an antibody provided herein comprises an Fc region with one or more amino acid substitutions at positions 239, 332, and 330, as described in Lazar et al., Proc. Natl. Acad. Sci. USA, 2006,103:4005-4010, incorporated by reference in its entirety.
[00216] Other illustrative glycosylation variants which may be incorporated into the antibodies provided herein are described, for example, in U.S. Pat. Pub. Nos. 2003/0157108, 2004/0093621, 2003/0157108, 2003/0115614, 2002/0164328, 2004/0093621, 2004/0132140, 2004/0110704, 2004/0110282, 2004/0109865; International Pat. Pub. Nos. 2000/61739, 2001/29246, 2003/085119, 2003/084570, 2005/035586, 2005/035778; 2005/053742, 2002/031140; Okazaki et al., J. Mol. Biol., 2004, 336: 1239-1249; and Yamane-Ohnuki et al., Biotech. Bioeng., 2004, 87: 614-622; each of which is incorporated by reference in its entirety.
[00217] In some embodiments, an antibody provided herein comprises an Fc region with at least one galactose residue in the oligosaccharide attached to the Fc region. Such antibody variants may have improved CDC function. Examples of such antibody variants are described, for example, in WO 1997/30087; WO 1998/58964; and WO 1999/22764; each of which his incorporated by reference in its entirety.
[00218] In some embodiments, an antibody provided herein comprises one or more alterations that improves or diminishes Clq binding and/or CDC. See U.S. Pat. No. 6,194,551; WO 99/51642; and Idusogie et al., J. Immunol., 2000, 164:4178-4184; each of which is incorporated by reference in its entirety.
Binding
[00219] The affinity of a molecule X for its partner Y can be represented by the dissociation equilibrium constant (KD). The kinetic components that contribute to the dissociation equilibrium constant are described in more detail below. Affinity can be measured by common methods known in the art, including those described herein, such as surface plasmon resonance (SPR) technology (e.g., BIACORE®) or biolayer interferometry (e.g., FORTEBIO®).
[00220] With regard to the binding of an antibody to a target molecule, the terms “bind,” “specific binding,” “specifically binds to,” “specific for,” “selectively binds,” and “selective for” a particular antigen (e.g., a polypeptide target) or an epitope on a particular antigen mean binding that is measurably different from a non-specific or non-selective interaction (e.g., with a non-target molecule). Specific binding can be measured, for example, by measuring binding to a target molecule and comparing it to binding to a non-target molecule. Specific binding can also be determined by competition with a control molecule that mimics the epitope recognized on the target molecule. In that case, specific binding is indicated if the binding of the antibody to the target molecule is competitively inhibited by the control molecule. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 50% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 40% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 30% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 20% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 10% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 1% of the affinity for fibrin. In some embodiments, the affinity of a fibrin antibody for a non-target molecule is less than about 0.1% of the affinity for fibrin.
[00221] When used herein in the context of two or more antibodies, the term “competes with” or “cross-competes with” indicates that the two or more antibodies compete for binding to an antigen (e.g., fibrin). In one exemplary assay, fibrin is coated on a surface and contacted with a first fibrin antibody, after which a second fibrin antibody is added. In another exemplary assay, a first fibrin antibody is coated on a surface and contacted with fibrin, and then a second fibrin antibody is added. If the presence of the first fibrin antibody reduces binding of the second fibrin antibody, in either assay, then the antibodies compete with each other. The term “competes with” also includes combinations of antibodies where one antibody reduces binding of another antibody, but where no competition is observed when the antibodies are added in the reverse order. However, in some embodiments, the first and second antibodies inhibit binding of each other, regardless of the order in which they are added. In some embodiments, one antibody reduces binding of another antibody to its antigen by at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% as measured in a competitive binding assay. A skilled artisan can select the concentrations of the antibodies used in the competition assays based on the affinities of the antibodies for fibrin and the valency of the antibodies. The assays described in this definition are illustrative, and a skilled artisan can utilize any suitable assay to determine if antibodies compete with each other. Suitable assays are described, for example, in Cox et al., “Immunoassay Methods,” in Assay Guidance Manual [Internet], Updated December 24, 2014 (ncbi.nlm.nih.gov/books/NBK92434/; accessed September 29, 2015); Silman et al., Cytometry, 2001, 44:30-37; and Finco et al., J. Pharm. Biomed. Anal., 2011, 54:351-358; each of which is incorporated by reference in its entirety.
[00222] A test antibody competes with a reference antibody if an excess of a test antibody (e.g., at least 2x, 5x, lOx, 20x, or lOOx) inhibits or blocks binding of the reference antibody by, e.g., at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% as measured in a competitive binding assay. Antibodies identified by competition assay (competing antibody) include antibodies binding to the same epitope as the reference antibody and antibodies binding to an adjacent epitope sufficiently proximal to the epitope bound by the reference antibody for steric hindrance to occur. For example, a second, competing antibody can be identified that competes for binding to fibrin with a first antibody described herein. In certain instances, the second antibody can block or inhibit binding of the first antibody by, e.g., at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% as measured in a competitive binding assay. In certain instances, the second antibody can displace the first antibody by greater than 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%. [00223] In some embodiments, an anti- fibrin antibody does not substantially bind myeloid cells present outside of cancer tissue. In some embodiments, an anti- fibrin antibody does not substantially bind stimulatory myeloid cells present in cancer tissue.
[00224] In some embodiments, an anti- fibrin antibody binds to residues y377-395 of the fibrin or fibrinogen yC domain (SEQ ID NO: 31) of human fibrin. The binding epitope includes the residues within the numerical range (e.g., residues 377-395 of fibrin), the beginning residue of each range (e.g., residues 377-394 of human fibrin) and the end residue of each range (e.g., residues 378-395 of human fibrin), or any combination thereof.
[00225] In some embodiments, an antibody provided herein binds human Fibrin with a KD of less than or equal to about 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.090, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 1.95, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, or lO x IO'6 M, as measured by ELISA or any other suitable method known in the art. In some embodiments, the KD of the antibody provided herein for the binding of human fibrin is between about 0.001-0.01, 0.01- 0.1, 0.01-0.05, 0.05-0.1, 0.1-0.5, 0.5-1, 0.25-0.75, 0.25-0.5, 0.5-0.75, 0.75-1, 0.75-2, 1.1-1.2, 1.2-1.3, 1.3-1.4, 1.4-1.5, 1.5-1.6, 1.6-1.7, 1.7-1.8, 1.8-1.9, 1.9-2, 1-2, 1-5, 2-7, 3-8, 3-5, 4-6, 5-7, 6-8, 7-9, 7-10, or 5-10xl0'6 M, as measured by ELISA or any other suitable method known in the art . In some embodiments, an antibody provided herein binds human Fibrin with a KD of less than or equal to about 1 x 10-8 M, or less than or equal to above 1 x 10'9 M as measured by ELISA or any other suitable method known in the art.
[00226] In some embodiments, the antibody provided herein binds human fibrin with a KD of less than or equal to about 10, 9, 8, 7, 6, 5, 4.5, 4, 3.5, 3, 2.5, 2, 1.98, 1.95, 1.9, 1.85, 1.8, 1.75, 1.7, 1.65, 1.6, 1.55, 1.50, 1.45, 1.4, 1.3, 1.2, 1.1, 1, 0.9, 0.85, 0.8, 0.75, 0.7, 0.65, 0.6, 0.55, 0.5, 0.45, 0.4, 0.35, 0.3, 0.25, 0.2, 0.15, 0.1, 0.05, 0.01, 0.005, 0.001, 0.0005, or 0.0001 x 10'8 M, or less, as measured by ELISA or any other suitable method known in the art . In some embodiments, the antibody provided herein binds human fibrin with a KD between 5-3, 4-2, 3-1, 1.9-1.8, 1.8-1.7, 1.7-1.6, 1.6-1.5, 1.9-1.5, 1.5-1, 1-0.8, 1-0.5, 0.9-0.6, 0.7-0.4, 0.6- 0.2, 0.5-0.3, 0.3-0.2, 0.2-0.1, 0.1-0.01, 0.01-0.001, or 0.001-0.0001 x IO'8 M as measured by ELISA or any other suitable method known in the art.
[00227] In some embodiments, an antibody provided herein binds human FGG with a KD of less than or equal to about 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, 0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 1.95, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, or 10 x 10'6 M, as measured by ELISA or any other suitable method known in the art. In some embodiments, the KD of the antibody provided herein for the binding of human FGG is between about 0.0001-0.001, 0.0001-0.0005, 0.0005-0.001, 0.001-0.01, 0.01-0.1, 0.01-0.05, 0.05-0.1, 0.1-0.5, 0.5-1, 0.25-0.75, 0.25-0.5, 0.5-0.75, 0.75-1, 0.75-2, 1.1-1.2, 1.2-1.3, 1.3- 1.4, 1.4-1.5, 1.5-1.6, 1.6-1.7, 1.7-1.8, 1.8-1.9, 1.9-2, 1-2, 1-5, 2-7, 3-8, 3-5, 4-6, 5-7, 6-8, 7-9, 7-10, or 5-10 x 10'8 M, as measured by ELISA or any other suitable method known in the art . In some embodiments, an antibody provided herein binds human FGG with a KD of less than or equal to about 1 x 10-8 M, or less than or equal to above 1 x 10-9 M as measured by ELISA or any other suitable method known in the art.
[00228] In some embodiments, an antibody provided herein binds P2 peptide with a KD of less than or equal to about 0.0001, 0.0002, 0.0003, 0.0004, 0.0005, 0.0006, 0.0007, 0.0008, 0.0009, 0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 1.95, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, or 10 x 10'6 M, as measured by ELISA or any other suitable method known in the art. In some embodiments, the KD of the antibody provided herein for the binding of P2 peptide is between about 0.0001-0.001, 0.0001-0.0005, 0.001-0.01, 0.01-0.1, 0.01-0.05, 0.05-0.1, 0.1- 0.5, 0.5-1, 0.25-0.75, 0.25-0.5, 0.5-0.75, 0.75-1, 0.75-2, 1.1-1.2, 1.2-1.3, 1.3-1.4, 1.4-1.5, 1.5- 1.6, 1.6-1.7, 1.7-1.8, 1.8-1.9, 1.9-2, 1-2, 1-5, 2-7, 3-8, 3-5, 4-6, 5-7, 6-8, 7-9, 7-10, or 5-10 x 10'6 M, as measured by ELISA or any other suitable method known in the art . In some embodiments, an antibody provided herein binds P2 peptide with a KD of less than or equal to about 1 x 10'8 M, or less than or equal to above 1 x 10'9 M as measured by ELISA or any other suitable method known in the art.
[00229] In some embodiments, an antibody provided herein binds fibrinogen with a KD of greater than or equal to about 1 x 10'6 M, greater than or equal to above 1 x 10'5 M, greater than or equal to above 1 x 10'4 M, or greater than or equal to above 1 x 10'3 M as measured by ELISA or any other suitable method known in the art.
[00230] In certain embodiments, the ratio of KD of the binding affinity of the antibody described herein to fibrin compared to fibrinogen is equal to or greater than 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 15, 20, 25,30, 35, 40, 45,50, 60, 70, 80, 90, 100, 150, 200, 300, 400, 500, or 1000.
Function
[00231] “Effector functions” refer to those biological activities mediated by the Fc region of an antibody, which activities may vary depending on the antibody isotype. Examples of antibody effector functions include receptor ligand blocking, agonism, or antagonism, Clq binding to activate complement dependent cytotoxicity (CDC), Fc receptor binding to activate antibody-dependent cellular cytotoxicity (ADCC), and antibody dependent cellular phagocytosis (ADCP). In some embodiments, the effector function of the fibrin antibody described herein is antagonism and blocks Mac-1 receptor binding to fibrin.
Pharmaceutical compositions
[00232] The present application provides compositions comprising the antibodies including pharmaceutical compositions comprising any one or more of the antibodies described herein with one or more pharmaceutically acceptable excipients. In some embodiments the composition is sterile. The pharmaceutical compositions generally comprise an effective amount of an antibody.
[00233] These compositions can comprise, in addition to one or more of the antibodies disclosed herein, a pharmaceutically acceptable excipient, carrier, buffer, stabilizer or other materials well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient. The precise nature of the carrier or other material can depend on the route of administration, e.g. oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular, intraperitoneal routes.
[00234] Pharmaceutical compositions for oral administration can be in tablet, capsule, powder or liquid form. A tablet can include a solid carrier such as gelatin or an adjuvant. Liquid pharmaceutical compositions generally include a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil. Physiological saline solution, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol can be included.
[00235] For intravenous, cutaneous or subcutaneous injection, or injection at the site of affliction, the active ingredient will be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has suitable pH, isotonicity and stability. Those of relevant skill in the art are well able to prepare suitable solutions using, for example, isotonic vehicles such as Sodium Chloride Injection, Ringer's Injection, Lactated Ringer's Injection. Preservatives, stabilisers, buffers, antioxidants and/or other additives can be included, as required.
[00236] The anti-fibrin antibody that is to be given to an individual, administration is preferably in a “therapeutically effective amount” or “prophylactically effective amount” (as the case can be, although prophylaxis can be considered therapy), this being sufficient to show benefit to the individual. The actual amount administered, and rate and time-course of administration, will depend on the nature and severity of protein aggregation disease being treated. Prescription of treatment, e.g. decisions on dosage etc., is within the responsibility of general practitioners and other medical doctors, and typically takes account of the disorder to be treated, the condition of the individual patient, the site of delivery, the method of administration and other factors known to practitioners. Examples of the techniques and protocols mentioned above can be found in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. (ed), 1980.
[00237] A composition can be administered alone or in combination with other treatments, either simultaneously or sequentially dependent upon the condition to be treated.
Methods
Methods of Preparation
[00238] Antibodies described herein can be produced using recombinant methods and compositions, e.g., as described in U.S. Pat. No. 4,816,567. In one embodiment, isolated nucleic acid encoding an antibody described herein is provided. Such nucleic acid may encode an amino acid sequence comprising the VL and/or an amino acid sequence comprising the VH of the antibody (e.g., the light and/or heavy chains of the antibody) or an amino acid sequence comprising the VHH of a single domain antibody. In a further embodiment, one or more vectors (e.g., expression vectors) comprising such nucleic acid are provided. In one embodiment, the nucleic acid is provided in a multi ci stronic vector. In a further embodiment, a host cell comprising such nucleic acid is provided. In one such embodiment, a host cell comprises (e.g., has been transformed with): (1) a vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antibody and an amino acid sequence comprising the VH of the antigen-binding polypeptide construct, or (2) a first vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antigen-binding polypeptide construct and a second vector comprising a nucleic acid that encodes an amino acid sequence comprising the VH of the antigen-binding polypeptide construct. In one embodiment, the host cell is eukaryotic, e.g. a Chinese Hamster Ovary (CHO) cell, or human embryonic kidney (HEK) cell, or lymphoid cell (e.g., Y0, NSO, Sp20 cell). In one embodiment, a method of making an antibody is provided, wherein the method comprises culturing a host cell comprising nucleic acid encoding the antibody, as provided above, under conditions suitable for expression of the antibody, and optionally recovering the antibody from the host cell (or host cell culture medium).
[00239] For recombinant production of the antibody, nucleic acid encoding an antibody, e.g., as described above, is isolated and inserted into one or more vectors for further cloning and/or expression in a host cell. Such nucleic acid may be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the antibody).
[00240] When a heteromultimer or variant thereof is recombinantly produced by the host cells, the protein in certain embodiments is present at about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% or less of the dry weight of the cells. When the heteromultimer or variant thereof is recombinantly produced by the host cells, the protein, in certain embodiments, is present in the culture medium at about 5 g/L, about 4 g/L, about 3 g/L, about 2 g/L, about 1 g/L, about 750 mg/L, about 500 mg/L, about 250 mg/L, about 100 mg/L, about 50 mg/L, about 10 mg/L, or about 1 mg/L or less of the dry weight of the cells. In certain embodiments, “substantially purified” heteromultimer produced by the methods described herein, has a purity level of at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, specifically, a purity level of at least about 75%, 80%, 85%, and more specifically, a purity level of at least about 90%, a purity level of at least about 95%, a purity level of at least about 99% or greater as determined by appropriate methods such as SDS/PAGE analysis, RP-HPLC, SEC, and capillary electrophoresis.
[00241] Suitable host cells for cloning or expression of antibody-encoding vectors include prokaryotic or eukaryotic cells described herein. [00242] Recombinant host cells or host cells are cells that include an exogenous polynucleotide, regardless of the method used for insertion, for example, direct uptake, transduction, f-mating, or other methods known in the art to create recombinant host cells. The exogenous polynucleotide may be maintained as a nonintegrated vector, for example, a plasmid, or alternatively, may be integrated into the host genome. Host cells can include CHO, derivatives of CHO, NSO, Sp2O, CV-1, VERO-76, HeLa, HepG2, Per.C6, or BHK.
[00243] For example, antibody may be produced in bacteria, in particular when glycosylation and Fc effector function are not needed. For expression of antibody fragments and polypeptides in bacteria, see, e.g., U.S. Pat. Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, N.J., 2003), pp. 245-254, describing expression of antibody fragments in E. coli.) After expression, the antibody may be isolated from the bacterial cell paste in a soluble fraction and can be further purified.
[00244] In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for antibody-encoding vectors, including fungi and yeast strains whose glycosylation pathways have been “humanized,” resulting in the production of an antibody with a partially or fully human glycosylation pattern. See Gerngross, Nat. Biotech. 22: 1409-1414 (2004), and Li et al., Nat. Biotech. 24:210-215 (2006).
[00245] Suitable host cells for the expression of glycosylated antibodies are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains have been identified which may be used in conjunction with insect cells, particularly for transfection of Spodoptera frugiperda cells.
[00246] Plant cell cultures can also be utilized as hosts. See, e.g., U.S. Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing PLANTIBODIES™ technology for producing antibodies in transgenic plants).
[00247] Vertebrate cells may also be used as hosts. For example, mammalian cell lines that are adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, e.g., in Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells as described, e.g., in Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK; buffalo rat liver cells (BRL 3 A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described, e.g., in Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines such as Y0, NS0 and Sp2/0. For a review of certain mammalian host cell lines suitable for antibody production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, N.J.), pp. 255-268 (2003).
[00248] In one embodiment, the antibodies described herein are produced in stable mammalian cells, by a method comprising: transfecting at least one stable mammalian cell with: nucleic acid encoding the antibody, in a predetermined ratio; and expressing the nucleic acid in the at least one mammalian cell. In some embodiments, the predetermined ratio of nucleic acid is determined in transient transfection experiments to determine the relative ratio of input nucleic acids that results in the highest percentage of the antibody in the expressed product.
[00249] In some embodiments, is the method of producing an antibody in stable mammalian cells as described herein wherein the expression product of the at least one stable mammalian cell comprises a larger percentage of the desired glycosylated antibody as compared to the monomeric heavy or light chain polypeptides, or other antibodies.
[00250] In some embodiments, is the method of producing a glycosylated antibody in stable mammalian cells described herein, said method comprising identifying and purifying the desired glycosylated antibody. In some embodiments, the said identification is by one or both of liquid chromatography and mass spectrometry.
[00251] If required, the antibodies can be purified or isolated after expression. Proteins may be isolated or purified in a variety of ways known to those skilled in the art. Standard purification methods include chromatographic techniques, including ion exchange, hydrophobic interaction, affinity, sizing or gel filtration, and reversed-phase, carried out at atmospheric pressure or at high pressure using systems such as FPLC and HPLC. Purification methods also include electrophoretic, immunological, precipitation, dialysis, and chromatofocusing techniques. Ultrafiltration and diafiltration techniques, in conjunction with protein concentration, are also useful. As is well known in the art, a variety of natural proteins bind Fc and antibodies, and these proteins can find use in the present invention for purification of antibodies. For example, the bacterial proteins A and G bind to the Fc region. Likewise, the bacterial protein L binds to the Fab region of some antibodies. Purification can often be enabled by a particular fusion partner. For example, antibodies may be purified using glutathione resin if a GST fusion is employed, Ni+2 affinity chromatography if a His-tag is employed or immobilized anti-flag antibody if a flag-tag is used. For general guidance in suitable purification techniques, see, e.g. incorporated entirely by reference Protein Purification: Principles and Practice, 3rd Ed., Scopes, Springer-Verlag, NY, 1994, incorporated entirely by reference. The degree of purification necessary will vary depending on the use of the antibodies. In some instances, no purification is necessary.
[00252] In certain embodiments, the antibodies are purified using Anion Exchange Chromatography including, but not limited to, chromatography on Q-sepharose, DEAE sepharose, poros HQ, poros DEAF, Toyopearl Q, Toyopearl QAE, Toyopearl DEAE, Resource/Source Q and DEAE, Fractogel Q and DEAE columns.
[00253] In specific embodiments, the proteins described herein are purified using Cation Exchange Chromatography including, but not limited to, SP-sepharose, CM sepharose, poros HS, poros CM, Toyopearl SP, Toyopearl CM, Resource/Source S and CM, Fractogel S and CM columns and their equivalents and comparables.
[00254] In addition, antibodies described herein can be chemically synthesized using techniques known in the art (e.g., see Creighton, 1983, Proteins: Structures and Molecular Principles, W. H. Freeman & Co., N.Y and Hunkapiller et al., Nature, 310: 105-111 (1984)). For example, a polypeptide corresponding to a fragment of a polypeptide can be synthesized by use of a peptide synthesizer. Furthermore, if desired, nonclassical amino acids or chemical amino acid analogs can be introduced as a substitution or addition into the polypeptide sequence. Non-classical amino acids include, but are not limited to, to the D-isomers of the common amino acids, 2,4diaminobutyric acid, alpha-amino isobutyric acid, 4aminobutyric acid, Abu, 2-amino butyric acid, g-Abu, e-Ahx, 6amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, alanine, fluoro-amino acids, designer amino acids such as methyl amino acids, C-methyl amino acids, N-methyl amino acids, and amino acid analogs in general. Furthermore, the amino acid can be D (dextrorotary) or L (levorotary).
Methods of Use
[00255] In an aspect, the present application provides methods of contacting fibrin with an anti-fibrin antibody, such as a human or humanized antibody, which results in inhibition of microglial adhesion to the fibrin or fibrinogen yC domain.
[00256] In an aspect, the present application provides methods of using the isolated antifibrin antibodies described herein for treatment of a degenerative disorder of the nervous system. In certain aspects, described herein is a method for treating a degenerative disorder of the nervous system, the method comprising administering to a mammalian subject a therapeutically effective amount of an anti-fibrin antibody or pharmaceutical composition comprising an anti-fibrin antibody described herein. In certain embodiments, the present application provides methods of treating a degenerative disorder of the nervous system selected from the group consisting of: multiple sclerosis, spinal cord injury, stroke, and Alzheimer’s Disease.
[00257] In certain aspects, described herein are methods for treating a pathology associated with Mac-1 binding to fibrin or Mac-1 binding with fibrinogen, the method comprising administering to a mammalian subject a therapeutically effective amount an isolated anti- fibrin antibody or a pharmaceutical composition comprising an isolated anti-fibrin antibody described herein.
[00258] In certain aspects, described herein are methods of inhibiting microglia activation, the method comprising administering to a mammalian subject a therapeutically effective amount an isolated anti-fibrin antibody or a pharmaceutical composition comprising an isolated antibody described herein.
[00259] In certain aspects, described herein is a method of preventing a degenerative disorder of the nervous system, the method comprising administering to a mammalian subject a therapeutically effective amount an isolated anti-fibrin antibody or a pharmaceutical composition comprising an isolated anti-fibrin antibody described herein. In certain embodiments, the present application provides methods of preventing a degenerative disorder of the nervous system selected from the group consisting of: multiple sclerosis, spinal cord injury, stroke, and Alzheimer’s Disease.
[00260] In certain aspects, described herein are methods of treating or preventing colitis.
Methods of Administration
[00261] In some embodiments, the methods provided herein are useful for the treatment of a degenerative nervous system disorder in an individual. In an embodiment, the individual is a human and the antibody is a fibrin antibody described herein.
[00262] In some embodiments, an antibody is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally. An effective amount of an anti-fibrin antibody may be administered for the treatment of cancer. The appropriate dosage of the anti-fibrin antibody may be determined based on the type of cancer to be treated, the type of the anti-fibrin antibody, the severity and course of the cancer, the clinical condition of the individual, the individual’s clinical history and response to the treatment, and the discretion of the attending physician.
[00263] In some embodiments, an antibody provided herein is administered with at least one additional therapeutic agent. Any suitable additional therapeutic or immunotherapeutic agent may be administered with an antibody provided herein. Additional therapeutic agents include agents that are used to treat or prevent a degenerative disorder of the nervous system selected from the group consisting of: multiple sclerosis, spinal cord injury, stroke, and Alzheimer’s Disease.
[00264] The additional therapeutic agent can be administered by any suitable means. In some embodiments, an antibody provided herein and the additional therapeutic agent are included in the same pharmaceutical composition. In some embodiments, an antibody provided herein and the additional therapeutic agent are included in different pharmaceutical compositions.
[00265] In embodiments where an antibody provided herein and the additional therapeutic agent are included in different pharmaceutical compositions, administration of the antibody can occur prior to, simultaneously, and/or following, administration of the additional therapeutic agent. In some embodiments, administration of an antibody provided herein and the additional therapeutic agent occur within about one month of each other. In some embodiments, administration of an antibody provided herein and the additional therapeutic agent occur within about one week of each other. In some embodiments, administration of an antibody provided herein and the additional therapeutic agent occur within about one day of each other. In some embodiments, administration of an antibody provided herein and the additional therapeutic agent occur within about twelve hours of each other. In some embodiments, administration of an antibody provided herein and the additional therapeutic agent occur within about one hour of each other.
Kits and Articles of Manufacture
[00266] The present application provides kits comprising any one or more of the antibody compositions described herein. In some embodiments, the kits further contain a component selected from any of secondary antibodies, reagents for immunohistochemistry analysis, pharmaceutically acceptable excipient and instruction manual and any combination thereof. In one specific embodiment, the kit comprises a pharmaceutical composition comprising any one or more of the antibody compositions described herein, with one or more pharmaceutically acceptable excipients.
[00267] The present application also provides articles of manufacture comprising any one of the antibody compositions or kits described herein. Examples of an article of manufacture include vials (including sealed vials).
EXAMPLES
[00268] Below are examples of specific embodiments for carrying out the present invention. The examples are offered for illustrative purposes only, and are not intended to limit the scope of the present invention in any way. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should, of course, be allowed for.
[00269] The practice of the present invention will employ, unless otherwise indicated, conventional methods of protein chemistry, biochemistry, recombinant DNA techniques and pharmacology, within the skill of the art. Such techniques are explained fully in the literature. See, e.g., T.E. Creighton, Proteins: Structures and Molecular Properties (W.H. Freeman and Company, 1993); A.L. Lehninger, Biochemistry (Worth Publishers, Inc., current addition); Sambrook, et al., Molecular Cloning: A Laboratory Manual (2nd Edition, 1989); Methods In Enzymology (S. Colowick and N. Kaplan eds., Academic Press, Inc.); Remington's Pharmaceutical Sciences, 18th Edition (Easton, Pennsylvania: Mack Publishing Company, 1990); Carey and Sundberg Advanced Organic Chemistry 3rd Ed. (Plenum Press) Vols A and B(1992).
Example 1: Phage display library screen for antibodies that bind Fibrin
[00270] Antibody screening using two human phage display libraries, a human hyperimmune Fab Library and a human structural scFv library, was performed by the following protocol. First, phage panning was performed, followed by phage enzyme-linked immunosorbent assays (ELISA) screening. Then, IgG reformatting and antibody production was performed followed by IgG charcterization by ELISA. Each library had three panning arms and 5 rounds of selection as shown in Figure 1. For positive selection, four reagents were used: Fibrin (thrombin cleaved fibrinogen), FGG (globular domain of Fibrinogen Gamma Chain), P2 peptide of FGG, and P13 peptide of Fibrinogen Beta chain (FGB). Positive selection began with 200 pmol antigen, and with each subsequent round of positive selection, the amount of antigen present was reduced by half. Wash stringency with PBS- Tween 20 (PBST) was also increased with each successive round of selection. For negative selection, Fibrinogen was used. Fibrinogen depletion was performed in Rounds 4 and 5 of selection. Figure 2 depicts the results of the phage display screen, where binding to FGG was increased significantly after each round of selection. 384 colonies were picked from round 5 and analysis by phage ELISA for binding to FGG and fibrinogen (Figure 2). Of the 384 clones selected, 33 clones were exhibited a ration of FGG/fibrinogen binding of greater than 2 as determined by ELISA (Fibure 2). 23 unique clones were identified from the hyperimmune Fab phage library and 24 unique clones were identified from the structural phage library (Figure 3), and these clones were reformatted into human IgG antibody format. LabChip® protein purity characterization, size exclusion chromatography and thermal stability analysis were performed on the reformatted, isolated antibody clones.
Example 2: Characterization of Binding Affinity of Reformatted Antibody Clones
[00271] IgG reformatted antibody clones obtained from the phage display screen as described in Example 1 were further characterized by ELISA for binding to Fibrin, FGG, P2 peptide, and Fibrinogen (Figures 4-10). Reformatted clones obtained from either the hyperimmune phage display antibody or structural scFv phage library displayed strong binding to fibrin and negligible binding to fibrinogen. [00272] A selected clone (TB241-03) was further examined for binding to recombinant biotinylated FGG having the F168A/F303A mutations (Figure 11). The protocol for expression and purification of FGG-F168A/F303A mutant was as performed as follows: On Day 0 - Cell preparation and transient transfection was performed. 293-BalanCD cells were seeded at 2.9 x 106 cells/ml in 293-BalanCD media in Erlenmeyer flasks. Incubator was set at: 6% CO2, 120 rpm, 37 deg. Transfection preparation was then preformed. The amounts were based on a 0.7 L transfection volume. 1 mg/ml cDNA concentration was used in 35 ml of OptiMEM media. 0.5 mg light chain and 0.5 mg heavy chain plasmid DNA was used. OptiMEM was transferred to two 50 ml Falcon tubes. 2.31 ml of PEI solution was added to one Falcon tube and 1 mg total cDNA to the other Falcon tube with OptiMEM media. The PEI/OptiMEM solution was added to sterile filtered DNA solution, mixed and incubated at RT for 25 min. The DNA/PEI/OptiMEM complex solution was added to the cell culture. On day 6, cell supernatant were harvested and sterile filetered. Protein purification was performed with Ni-NTA resin to capture His-tagged FGG protein. The sterilized supernatant was added to a 10 ml Ni-NTA column. The Ni-NTA column was washed with 10 CV of wash buffer (20 mM Tris, 500 mM NaCl, 20 mM Imidazole, pH 8). His tagged FGG protein was eluted with 5 CV of Elution buffer (20 mM Tris, 150 mM NaCl, 250 mM Imidazole, pH 8). The eluate was then dialyzed into lx PBS for storage at -80 deg. and protein analysis.
[00273] The Gator protocol was then performed to study the binding of reformatted antibodies to the recombinant, purified FGG-F168A/F303A mutant. Material used: Gator/Octet (Cat# GA00051), Streptavidin sensors (Cat # 160002), K-buffer (Cat # 120011)
[00274] Ligand - FGG-His mutant-biotinylated and loaded on streptavidin probes- 400nM solution (30.2uM, MW= 29.8 KDa). 500nM working solution of the following: purified antibody clone TB241-02 at stock concentration 10.67uM, purified antibody clone TB241-03 at stock concentration 7.33uM, purified antibody clone TB263-02 at stock concentration 4uM, purified antibody clone TB241-02-lala at stock concentration 6.67uM, purified antibody clone TB239-02 at stock concentration lOuM. The buffers used were as follows: K Buffer (40C)- 1 :10 dilution of Q Buffer(40C) in IX PBS and 0.01% Pluronic, that was used for dilution of analyte and to establish baseline. The parameters used on the Gator were seta as: 5Hz, 96 well plate, tilt, 300C. [00275] Figure 11 depicts the results of selected clones binding to recombinant biotinylated - FGG F168A/F303A, demonstrating that clone TB239-02 binds strongly to the recombinant mutant FGG
[00276] These results confirm that the antibody clones bind to fibrin preferentially over fibrinogen.
Table D: Binding Characteristics of Selected Clones
Fibrin EC5O FGG EC5O P2 EC5O Fibrinogen Panning
Clone Name Library (nM) (nM) (nM) EC5O (nM) Arm
Structural
TB241-04 0.7 7.0 no data x Fibrin/P13 scFv Structural
TB240-05 0.3 I .6 4.9 x Fibrin/P2 scFv Structural
TB240-04 x 12.1 x X Fibrin/P2 scFv Structural
TB239-03 1.2 9.2 no data X Fibrin/FGG scFv Structural
TB239-02 1.3 I I .5 no data X Fibrin/FGG scFv Hyperimmune
TB237-11 0.4 1.5 8.6 X Fibrin/P2 Fab Structural
TB240-03 2.3 13.6 70.4 X Fibrin/P2 scFv Structural
TB240-02 1.9 29.7 54.5 X Fibrin/P2 scFv Structural
TB241-05 2.7 x no data X Fibrin/P13 scFv Structural
TB241-03 19.1 x no data X Fibrin/P13 scFv Structural
TB241-02 0.8 4.5 no data X Fibrin/P13 scFv Structural
TB239-09 x x no data X Fibrin/FGG scFv Structural
TB239-10 x X no data X Fibrin/FGG scFv Structural
TB239-06 3.8 X no data X Fibrin/FGG scFv
2.0 7.8 Structural
TB239-05 no data X Fibrin/FGG scFv Structural
TB239-12 x x no data X Fibrin/FGG scFv Hyperimmune
TB237-12 0.3 1.1 5.1 X Fibrin/P2 Fab Structural
TB239-07 3.4 6.9 no data X Fibrin/FGG scFv Hyperimmune
TB236-02 0.3 1.5 5.8 X Fibrin/FGG Fab 4 Hyperimmune
TB237-02 0.2 0.8 4. X Fibrin/P2 Fab Hyperimmune
TB237-09 0.7 2.3 9.9 X Fibrin/P2 Fab Hyperimmune
TB238-02 0.5 2.9 no data X Fibrin/P13 Fab Hyperimmune
TB237-07 0.4 3.7 14.0 X Fibrin/P2 Fab
TB238-05 1.4 6.6 no data X Hyperimmune Fibrin/P13 Fab
Structural
TB239-14 3.1 x no data X Fibrin/FGG scFv Hyperimmune
TB237-04 0.3 1.3 4.6 X Fibrin/P2 Fab Hyperimmune
TB237-14 18.4 X X X Fibrin/P2 Fab Hyperimmune
TB238-03 0.3 1 .7 no data X Fibrin/P13 Fab Structural
TB240-01 0.3 2.5 16.1 X Fibrin/P2 scFv Structural
TB241-01 0.3 1 .8 no data x Fibrin/P13 scFv Hyperimmune
TB237-O3 0.1 0.6 4.8 X Fibrin/P2 Fab Hyperimmune
TB237-O6 0.5 2.3 9.6 X Fibrin/P2 Fab Structural
TB239-O1 x X no data X Fibrin/FGG scFv Structural
TB239-13 1 .9 4.5 no data x Fibrin/FGG scFv Hyperimmune
TB236-O1 0.4 1.1 3.8 X Fibrin/FGG Fab Hyperimmune
TB238-O7 0.5 1 .5 no data x Fibrin/P13 Fab Hyperimmune
TB237-O1 0.5 1.2 8.0 X Fibrin/P2 Fab Hyperimmune
TB237-O5 2.0 12.3 34.6 X Fibrin/P2 Fab Hyperimmune
TB237-1O 0.3 1.4 6.0 X Fibrin/P2 Fab Hyperimmune
TB237-13 0.4 2.7 1 1 .5 X Fibrin/P2 Fab Hyperimmune
TB237-O8 2.9 5.1 20.1 X Fibrin/P2 Fab Structural
TB239-O8 0.3 0.9 no data x Fibrin/FGG scFv Hyperimmune
TB238-O1 0.3 1 .0 no data x Fibrin/P13 Fab
Example 3: Anti-Fibrin antibody delays and reduces clinical symptoms of disease in rodent EAE model of Multiple Sclerosis
[00277] The ability of the anti-fibrin antibodies to treat Multiple Sclerosis was assessed using a mouse model of experimental autoimmune encephalomyelitis (EAE) (Figure 12). SJL female mice were induced with EAE disease by injection of PLP131-151 peptide 2 hours post-injection, mice were immunized with a single injection of 60 ng/doseof Pertussis toxin. Antibody clone TB239-02 was dosed at 15 mg/Kg on days 0 and 1 after immunization.
Animals were monitored and scored for onset of paralysis (Figure 12). Mice treated with TB239-02 demonstrated a delay in onset of paralysis and a reduction of clinical score as compared to vehicle or isotype controls. [00278] These results demonstrate that the antibody clones described herein can be used for the treatment of Multiple Sclerosis.
Example 4: Treatment of neurodegenerative disaese with anti-Fibrin antibodies
[00279] The purified antibody clones described herein are formulated into a pharmaceutical composition to be administered to patients for the treatment of a neurodegenerative disease (e.g., multiple sclerosis or Alzheimer’s Disease). The pharmaceutical composition comprising an antibody clonedescribed herein is administered at a dose sufficient to effectively reduce the symptoms of the neurodegenerative disease. The pharmaceutical composition is well tolerated and does not induce significant harmful adverse effects in the patient.
Example 5: Treatment of colitis with anti-Fibrin antibodies
[00280] The antibody clones described herein are investigated the ability to treat colitis in a mouse model of colitis. To initiate dextran sodium sulfate (DSS) induced colitis treatment, 8- 10 week old female C57BL/6 mice are acclimated to the vivarium for at least 4 days, weighed, and randomized into treatment groups based on body weight. 2 types of studies are conducted: acute (7 days) and chronic (28 days).
[00281] The acute DSS study is performed by adding 2.5% DSS into drinking water for 7 days. Antibodies are administered IP every 2 days (Q2D) at 10 and 30mg/kg. Mice are euthanized on day 7 with isoflurane anesthesia, exsanguination, followed by cervical dislocation. The colon is removed and analyzed for histopathology.
[00282] The chronic DSS study is performed by adding 2.0% DSS into drinking water for 1 week followed by replacing with 1 week of normal drinking water, followed by another week of 2% DSS and ending with another week of normal drinking water. Antibody clones described herein are administered IV prophylactically starting on day 0 twice a week at 30 and 5 mg/kg. Mice are euthanized after 28 days with isoflurane anesthesia, exsanguination, followed by cervical dislocation. The colon is removed and analyzed for histopathology.
[00283] These results confirm that the antibody clones described herein are effective for the treatment of colitis.
[00284] While the invention has been particularly shown and described with reference to a preferred embodiment and various alternate embodiments, it will be understood by persons skilled in the relevant art that various changes in form and details can be made therein without departing from the spirit and scope of the invention.
[00285] All references, issued patents and patent applications cited within the body of the instant specification are hereby incorporated by reference in their entirety, for all purposes.
INFORMAL SEQUENCE LISTING
Figure imgf000091_0001
Figure imgf000092_0001
Figure imgf000093_0001
Figure imgf000094_0001
Figure imgf000095_0001
Figure imgf000096_0001
Figure imgf000097_0001
Figure imgf000098_0001
Figure imgf000099_0001
Figure imgf000100_0001
Figure imgf000101_0001
Figure imgf000102_0001
Figure imgf000103_0001
Figure imgf000104_0001
Figure imgf000105_0001
Figure imgf000106_0001
Figure imgf000107_0001
Figure imgf000108_0001
Figure imgf000109_0001
Figure imgf000110_0001
Figure imgf000111_0001
Figure imgf000112_0001
Figure imgf000113_0001
Figure imgf000114_0001
Figure imgf000115_0001
Figure imgf000116_0001
Figure imgf000117_0001
Figure imgf000118_0001
Figure imgf000119_0001
Figure imgf000120_0001
Figure imgf000121_0001
Figure imgf000122_0001
Figure imgf000123_0001
Figure imgf000124_0001
Figure imgf000125_0001
Figure imgf000126_0001

Claims

1. An isolated antibody that binds human fibrin, comprising a heavy chain comprising a variable heavy (VH) chain sequence comprising three heavy chain CDR sequences, CDR-H1, CDR-H2, and CDR-H3, and a light chain comprising a variable light (VL) chain sequence comprising three light chain CDR sequences, CDR-L1, CDR-L2, and CDR-L3, wherein: a. CDR-H1 comprises the sequence set forth in SEQ ID NO: 1, 13, 25, 37, 49,
61, 73, 85, 97, 109, 121, 133, 145, 157, 169, 181, 193, 205, 217, 229, 244,
256, 268, 280, 292, 304, 316, 328, 340, 352, 364, 376, 388, 400, 412, 424,
436, 448, 460, 472, 484, 496, 508, 520, 532, 546, or 556; b. CDR-H2 comprises the sequence set forth in SEQ ID NO: 2, 14, 26, 38, 50,
62, 74, 86, 98, 110, 122, 134, 146, 158, 170, 182, 194, 206, 218, 230, 245,
257, 269, 281, 293, 305, 317, 329, 341, 353, 365, 378, 389, 401, 413, 425,
437, 449, 461, 473, 485, 497, 509, 521, 533, 547, or 557; c. CDR-H3 comprises the sequence set forth in SEQ ID NO: 3, 15, 27, 39, 51,
63, 75, 87, 99, 111, 123, 135, 147, 159, 171, 183, 195, 207, 219, or 231, 246,
258, 270, 282, 294, 306, 318, 330, 342, 354, 366, 379, 390, 402, 414, 426,
438, 450, 462, 474, 486, 498, 510, 522, 534, 548, or 558; d. CDR-L1 comprises the sequence set forth in SEQ ID NO: 4, 16, 28, 40, 52,
64, 76, 88, 100, 112, 124, 136, 148, 160, 172, 184, 196, 208, 220, 232, 247,
259, 271, 283, 295, 307, 319, 331, 343, 355, 367, 380, 391, 403, 415, 427,
439, 451, 463, 475, 487, 499, 511, 523, 535, 549, or 559; e. CDR-L2 comprises the sequence set forth in SEQ ID NO: 5, 17, 29, 41, 53,
65, 77, 89, 101, 113, 125, 137, 149, 161, 173, 185, 197, 209, 221, 233, 248,
260, 272, 284, 296, 308, 320, 332, 344, 356, 368, 381, 392, 404, 416, 428,
440, 452, 464, 476, 488, 500, 512, 524, 536, 550, or 560; and f. CDR-L3 comprises the sequence set forth in SEQ ID NO: 6, 18, 30, 42, 54,
66, 78, 90, 102, 114, 126, 138, 150, 162, 174, 186, 198, 210, 222, 234, 249,
261, 273, 285, 297, 309, 321, 333, 345, 357, 369, 382, 393, 405, 417, 429,
441, 453, 465, 477, 489, 501, 513, 525, 537, 551, or 561.
2. The isolated antibody of claim 1, wherein the antibody comprises a VH sequence selected from a sequence set forth in one of SEQ ID NOs: 7, 19, 31, 43, 55, 67, 79, 91, 103, 115, 127, 139, 151, 163, 175, 187, 199, 211, 223, 235, 250, 262, 274, 286, 298, 310, 322, 334, 346, 358, 370, 382, 394, 406, 418, 430, 442, 454, 466, 478, 490, 502, 514, 526, 538, 550, or 562.
3. The isolated antibody of claim 1 or 2, wherein the antibody comprises a VL sequence selected from a sequence set forth in SEQ ID NOs: 10, 22, 34, 46, 58, 70, 82, 94, 106, 118, 130, 142, 154, 166, 178, 190, 202, 214, 226, 238, 253, 265, 277, 289, 301, 313, 325, 337, 349, 361, 373, 385, 397, 409, 421, 433, 445, 457, 469, 481, 493, 505, 517, 529, 541, 553, or 565.
4. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence selected from a sequence set forth in one of SEQ ID Nos: 7, and the VL sequence set for in SEQ ID NO: 10.
5. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 19 and a VL sequence set forth in SEQ ID NO: 22.
6. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 31 and a VL sequence set forth in SEQ ID NO: 34.
7. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 43 and a VL sequence set forth in SEQ ID NO: 46.
8. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 55 and a VL sequence set forth in SEQ ID NO: 58.
9. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 67 and a VL sequence set forth in SEQ ID NO: 70.
10. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 79 and a VL sequence set forth in SEQ ID NO: 82.
11. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 91 and a VL sequence set forth in SEQ ID NO: 94.
12. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 103 and a VL sequence set forth in SEQ ID NO: 106.
13. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 115 and a VL sequence set forth in SEQ ID NO: 118.
14. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 127 and a VL sequence set forth in SEQ ID NO: 130.
15. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 139 and a VL sequence set forth in SEQ ID NO: 142.
16. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 151 and a VL sequence set forth in SEQ ID NO: 154.
17. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 163 and a VL sequence set forth in SEQ ID NO: 166.
18. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 175 and a VL sequence set forth in SEQ ID NO: 178.
19. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 187 and a VL sequence set forth in SEQ ID NO: 190.
20. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 199 and a VL sequence set forth in SEQ ID NO: 202.
21. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 211 and a VL sequence set forth in SEQ ID NO: 214.
22. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 223 and a VL sequence set forth in SEQ ID NO: 226.
23. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 235 and a VL sequence set forth in SEQ ID NO: 238.
24. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 250, and a VL sequence set forth in SEQ ID NO: 253.
25. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 262 and a VL sequence set forth in SEQ ID NO: 265.
26. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 274 and a VL sequence set forth in SEQ ID NO: 277.
27. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 286 and a VL sequence set forth in SEQ ID NO: 289.
28. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 298 and a VL sequence set forth in SEQ ID NO: 301.
29. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 310, and a VL sequence set forth in SEQ ID NO: 313.
30. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 322 and a VL sequence set forth in SEQ ID NO: 325.
31. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 334 and a VL sequence set forth in SEQ ID NO: 337.
32. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 346 and a VL sequence set forth in SEQ ID NO: 349.
33. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 358 and a VL sequence set forth in SEQ ID NO: 361.
34. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 370 and a VL sequence set forth in SEQ ID NO: 373.
35. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 382 and a VL sequence set forth in SEQ ID NO: 385.
36. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 394 and a VL sequence set forth in SEQ ID NO: 397.
37. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 406 and a VL sequence set forth in SEQ ID NO: 409.
38. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 418 and a VL sequence set forth in SEQ ID NO: 421.
39. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 430 and a VL sequence set forth in SEQ ID NO: 433.
40. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 442 and a VL sequence set forth in SEQ ID NO: 445.
128
41. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 454 and a VL sequence set forth in SEQ ID NO: 457.
42. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 466 and a VL sequence set forth in SEQ ID NO: 469.
43. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 478 and a VL sequence set forth in SEQ ID NO: 481.
44. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 490 and a VL sequence set forth in SEQ ID NO: 493.
45. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 502 and a VL sequence set forth in SEQ ID NO: 505.
46. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 514 and a VL sequence set forth in SEQ ID NO: 517.
47. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 526 and a VL sequence set forth in SEQ ID NO: 529.
48. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 538 and a VL sequence set forth in SEQ ID NO: 541.
49. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 550 and a VL sequence set forth in SEQ ID NO: 553.
50. The isolated antibody of any one of claims 1-3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 562 and a VL sequence set forth in SEQ ID NO: 565.
51. The isolated antibody of any one of the above claims, wherein the antibody is a humanized, human or chimeric antibody.
52. The isolated antibody of claim 25, wherein the antibody is a humanized antibody.
53. The isolated antibody of any one of the above claims, wherein the antibody comprises a heavy chain human constant region of a class selected from IgG, IgA, IgD, IgE, and IgM.
54. The isolated antibody of any one of the above claims, wherein the human Fc region comprises a human heavy chain constant region of the class IgG and a subclass selected from IgGl, IgG2, IgG3, and IgG4.
129
55. The isolated antibody of claim 54, wherein the human Fc region comprises wild-type, human IgGl Fc.
56. The isolated antibody of claim 55, wherein the human Fc domain comprises a sequence set forth in SEQ ID NO: 8, 20, 32, 44, 56, 68, 80, 92, 104, 116, 128, 140, 152, 164, 176, 188, 200, 212, 224, 236, 251, 263, 275, 287, 299, 311, 323, 335, 347, 359, 371, 383, 395, 407, 419, 431, 443, 455, 467, 479, 491, 503, 515, 527, 539, 551, or 563.
57. The isolated antibody of any one of the above claims, wherein the heavy chain comprises a constant heavy chain sequence set forth by SEQ ID NO: 8, 20, 32, 44, 56, 68, 80,
92, 104, 116, 128, 140, 152, 164, 176, 188, 200, 212, 224, 236, 251, 263, 275, 287, 299, 311,
323, 335, 347, 359, 371, 383, 395, 407, 419, 431, 443, 455, 467, 479, 491, 503, 515, 527,
539, 551, or 563.
58. The isolated antibody of any one of the above claims, wherein the light chain comprises a constant light chain sequence set forth by SEQ ID NO: 9, 21, 33, 45, 57, 69, 81,
93, 105, 117, 129, 141, 153, 165, 177, 189, 201, 213, 225, 237, 252, 264, 276, 288, 300, 312,
324, 336, 348, 360, 372, 384, 396, 408, 420, 432, 444, 456, 468, 480, 492, 504, 516, 528,
540, 552, or 564.
59. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 7, and the VL sequence set forth in SEQ ID NO: 10; and the human Fc region comprises wild-type, human IgGl Fc.
60. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 19, and the VL sequence set forth in SEQ ID NO: 22; and the human Fc region comprises wild-type, human IgGl Fc.
61. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 31, and the VL sequence set forth in SEQ ID NO: 34; and the human Fc region comprises wild-type, human IgGl Fc.
62. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 43, and the VL sequence set forth in SEQ ID NO: 46; and the human Fc region comprises wild-type, human IgGl Fc.
63. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 55, and the VL sequence set forth in SEQ ID NO: 58; and the human Fc region comprises wild-type, human IgGl Fc.
130
64. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 67, and the VL sequence set forth in SEQ ID NO: 70; and the human Fc region comprises wild-type, human IgGl Fc.
65. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 79, and the VL sequence set forth in SEQ ID NO: 82; and the human Fc region comprises wild-type, human IgGl Fc.
66. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 91, and the VL sequence set forth in SEQ ID NO: 94; and the human Fc region comprises wild-type, human IgGl Fc.
67. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 103, and the VL sequence set forth in SEQ ID NO: 106; and the human Fc region comprises wild-type, human IgGl Fc.
68. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 115, and the VL sequence set forth in SEQ ID NO: 118; and the human Fc region comprises wild-type, human IgGl Fc.
69. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 127, and the VL sequence set forth in SEQ ID NO: 130; and the human Fc region comprises wild-type, human IgGl Fc.
70. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 139, and the VL sequence set forth in SEQ ID NO: 142; and the human Fc region comprises wild-type, human IgGl Fc.
71. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 151, and the VL sequence set forth in SEQ ID NO: 154; and the human Fc region comprises wild-type, human IgGl Fc.
72. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 163, and the VL sequence set forth in SEQ ID NO: 166; and the human Fc region comprises wild-type, human IgGl Fc.
73. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 175, and the VL sequence set forth in SEQ ID NO: 178; and the human Fc region comprises wild-type, human IgGl Fc.
131
74. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 187, and the VL sequence set forth in SEQ ID NO: 190; and the human Fc region comprises wild-type, human IgGl Fc.
75. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 199, and the VL sequence set forth in SEQ ID NO: 202; and the human Fc region comprises wild-type, human IgGl Fc.
76. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 211, and the VL sequence set forth in SEQ ID NO: 214; and the human Fc region comprises wild-type, human IgGl Fc.
77. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 223, and the VL sequence set forth in SEQ ID NO: 226; and the human Fc region comprises wild-type, human IgGl Fc.
78. The isolated antibody of any one of the above claims, wherein the antibody comprises the VH sequence set forth in SEQ ID NO: 235, and the VL sequence set forth in SEQ ID NO: 238; and the human Fc region comprises wild-type, human IgGl Fc.
79. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 250, and a VL sequence set forth in SEQ ID NO: 253; and the human Fc region comprises wild-type, human IgGl Fc.
80. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 262 and a VL sequence set forth in SEQ ID NO: 265; and the human Fc region comprises wild-type, human IgGl Fc..
81. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 274 and a VL sequence set forth in SEQ ID NO: 277; and the human Fc region comprises wild-type, human IgGl Fc.
82. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 286 and a VL sequence set forth in SEQ ID NO: 289; and the human Fc region comprises wild-type, human IgGl Fc.
83. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 298 and a VL sequence set forth in SEQ ID NO: 301; and the human Fc region comprises wild-type, human IgGl Fc.
132
84. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 310, and a VL sequence set forth in SEQ ID NO: 313; and the human Fc region comprises wild-type, human IgGl Fc.
85. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 322 and a VL sequence set forth in SEQ ID NO: 325; and the human Fc region comprises wild-type, human IgGl Fc.
86. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 334 and a VL sequence set forth in SEQ ID NO: 337; and the human Fc region comprises wild-type, human IgGl Fc.
87. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 346 and a VL sequence set forth in SEQ ID NO: 349; and the human Fc region comprises wild-type, human IgGl Fc.
88. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 358 and a VL sequence set forth in SEQ ID NO: 361; and the human Fc region comprises wild-type, human IgGl Fc.
89. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 370 and a VL sequence set forth in SEQ ID NO: 373; and the human Fc region comprises wild-type, human IgGl Fc.
90. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 382 and a VL sequence set forth in SEQ ID NO: 385; and the human Fc region comprises wild-type, human IgGl Fc.
91. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 394 and a VL sequence set forth in SEQ ID NO: 397; and the human Fc region comprises wild-type, human IgGl Fc.
92. The isolated antibody of any one of the above claims 3, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 406 and a VL sequence set forth in SEQ ID NO: 409; and the human Fc region comprises wild-type, human IgGl Fc.
93. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 418 and a VL sequence set forth in SEQ ID NO: 421; and the human Fc region comprises wild-type, human IgGl Fc.
133
94. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 430 and a VL sequence set forth in SEQ ID NO: 433; and the human Fc region comprises wild-type, human IgGl Fc.
95. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 442 and a VL sequence set forth in SEQ ID NO: 445; and the human Fc region comprises wild-type, human IgGl Fc.
96. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 454 and a VL sequence set forth in SEQ ID NO: 457; and the human Fc region comprises wild-type, human IgGl Fc.
97. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 466 and a VL sequence set forth in SEQ ID NO: 469; and the human Fc region comprises wild-type, human IgGl Fc.
98. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 478 and a VL sequence set forth in SEQ ID NO: 481; and the human Fc region comprises wild-type, human IgGl Fc.
99. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 490 and a VL sequence set forth in SEQ ID NO: 493; and the human Fc region comprises wild-type, human IgGl Fc.
100. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 502 and a VL sequence set forth in SEQ ID NO: 505; and the human Fc region comprises wild-type, human IgGl Fc.
101. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 514 and a VL sequence set forth in SEQ ID NO: 517; and the human Fc region comprises wild-type, human IgGl Fc.
102. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 526 and a VL sequence set forth in SEQ ID NO: 529; and the human Fc region comprises wild-type, human IgGl Fc.
103. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 538 and a VL sequence set forth in SEQ ID NO: 541; and the human Fc region comprises wild-type, human IgGl Fc.
134
104. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 550 and a VL sequence set forth in SEQ ID NO: 553; and the human Fc region comprises wild-type, human IgGl Fc.
105. The isolated antibody of any one of the above claims, wherein the antibody comprises a VH sequence set forth in SEQ ID NO: 562 and a VL sequence set forth in SEQ ID NO: 565; and the human Fc region comprises wild-type, human IgGl Fc.
106. The isolated antibody of any one of claims 54 - 105, wherein the Fc region comprises one or more amino acid substitutions, wherein the one or more substitutions result in increased antibody half-life, increased ADCC activity, increased ADCP activity, or increased CDC activity compared with the Fc without the one or more substitutions.
107. The isolated antibody of any one of the above claims, wherein the Fc region binds an Fey Receptor selected from the group consisting of: FcyRI, FcyRIIa, FcyRIIb, FcyRIIc, FcyRIIIa, and FcyRIIIb.
108. The isolated antibody of any one of the above claims, wherein the Fc region has reduced or is lacking Fc effector function compared to a wild-type Fc region.
109. The isolated antibody of any one of the above claims, wherein the antibody is a monoclonal antibody.
110. The antibody of any one of the above claims, wherein the antibody binds Fibrin with a KD of less than or equal to about 1 xlO'8M, as measured by enzyme-linked immunosorbent assay (ELISA).
111. The antibody of any one of the above claims, wherein the antibody binds Fibrin Gamma Globular (FGG) domain.
112. The antibody of any one of the above claims, wherein the antibody binds FGG with a KD of less than or equal to about 1 xlO'8M, as measured by ELISA.
113. The isolated antibody of any one of the above claims, wherein the antibody binds to a peptide comprising SEQ ID NO: 241 with a KD of less than or equal to about 1 xlO'8M, as measured by ELISA.
114. The isolated antibody of any one of the above claims, wherein the antibody binds to fibrinogen with a KD of greater than or equal to about 1 xlO'6M, as measured by ELISA.
135
115. The isolated antibody of any one of the above claims, wherein the antibody inhibits Mac-1 binding to fibrin or fibrinogen yC domain.
116. The isolated antibody of any one of the above claims, wherein the antibody exhibits inhibition of microglial adhesion to the fibrin or fibrinogen yC domain.
117. The isolated antibody of any one of the above claims, wherein the antibody exhibits activation of CD1 IB positive monocytic cells.
118. The isolated antibody of any one of the above claims for use in the treatment of a degenerative disorder of the nervous system.
119. An isolated polynucleotide or set of polynucleotides encoding the antibody of any of the above claims, a VH thereof, a VL thereof, a light chain thereof, a heavy chain thereof, or an antigen-binding portion thereof, and optionally, wherein the polynucleotide or set of polynucleotides comprises cDNA.
120. A vector or set of vectors comprising the polynucleotide or set of polynucleotides of claim 119.
121. A host cell comprising the polynucleotide or set of polynucleotides of claim 119 or the vector or set of vectors of claim 120.
122. A method of producing an antibody, the method comprising expressing the antibody with the host cell of claim 121 and isolating the expressed antibody.
123. A pharmaceutical composition comprising the antibody of any one of claims 1 - 118 and a pharmaceutically acceptable excipient.
124. A kit comprising the antibody of any one of claims 1 - 118 or a pharmaceutical composition of claim 123 and instructions for use.
125. A method for treating a degenerative disorder of the nervous system, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody of any one of claims 1 - 118 or a pharmaceutical composition of claim 123.
126. The method of claim 125, wherein the degenerative disorder of the nervous system is selected from the group consisting of: multiple sclerosis, spinal cord injury, stroke, and Alzheimer’s Disease.
136
127. A method of treating a pathology associated with Mac-1 binding to fibrin or Mac-1 binding with fibrinogen, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody of any one of claims 1 - 118 or a pharmaceutical composition of claim 123.
128. A method of inhibiting microglia activation, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody of any one of claims 1 - 118 or a pharmaceutical composition of claim 123.
129. A method of preventing a degenerative disorder of the nervous system, the method comprising administering to a mammalian subject a therapeutically effective amount of the antibody of any one of claims 1 - 118 or a pharmaceutical composition of claim 123.
130. A method of treating colitis in a subject in need thereof, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody of any one of claims 1 - 118 or a pharmaceutical composition of claim 123.
131. A method of preventing colitis in a subject in need thereof, the method comprising administering to a mammalian subject a therapeutically effective amount the antibody of any one of claims 1 - 118 or a pharmaceutical composition of claim 123.
PCT/US2022/013423 2021-01-21 2022-01-21 Antibodies which bind human fibrin and methods of use WO2022159776A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202163140138P 2021-01-21 2021-01-21
US63/140,138 2021-01-21

Publications (1)

Publication Number Publication Date
WO2022159776A1 true WO2022159776A1 (en) 2022-07-28

Family

ID=82549241

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2022/013423 WO2022159776A1 (en) 2021-01-21 2022-01-21 Antibodies which bind human fibrin and methods of use

Country Status (1)

Country Link
WO (1) WO2022159776A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024044583A1 (en) * 2022-08-22 2024-02-29 The J. David Gladstone Institutes, A Testamentary Trust Established Under The Will Of J. David Gladstone Human anti-fibrin antibodies and uses thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5821068A (en) * 1993-11-02 1998-10-13 Iatron Laboratories, Inc. Anti-human soluble fibrin antibody, hybridoma, and immunoassaying method
WO2003000736A1 (en) * 2001-06-26 2003-01-03 Agen Biomedical Limited Humanized antibodies derived from dd-3b6/22, specific for the d-dimer fragment of fibrin
WO2011041518A1 (en) * 2009-10-02 2011-04-07 The Regents Of The University Of California Monoclonal antibodies

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5821068A (en) * 1993-11-02 1998-10-13 Iatron Laboratories, Inc. Anti-human soluble fibrin antibody, hybridoma, and immunoassaying method
WO2003000736A1 (en) * 2001-06-26 2003-01-03 Agen Biomedical Limited Humanized antibodies derived from dd-3b6/22, specific for the d-dimer fragment of fibrin
WO2011041518A1 (en) * 2009-10-02 2011-04-07 The Regents Of The University Of California Monoclonal antibodies

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
RYU JAE KYU, RAFALSKI VICTORIA A., MEYER-FRANKE ANKE, ADAMS RYAN A., PODA SURESH B., RIOS CORONADO PAMELA E., PEDERSEN LARS ØSTERG: "Fibrin-targeting immunotherapy protects against neuroinflammation and neurodegeneration", NATURE IMMUNOLOGY, vol. 19, no. 11, 15 April 2019 (2019-04-15), pages 1212 - 1223, XP036617636 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024044583A1 (en) * 2022-08-22 2024-02-29 The J. David Gladstone Institutes, A Testamentary Trust Established Under The Will Of J. David Gladstone Human anti-fibrin antibodies and uses thereof

Similar Documents

Publication Publication Date Title
CN107849136B (en) anti-TfR antibodies and their use in the treatment of proliferative and inflammatory diseases
CA3081125A1 (en) Fully humanized anti-b cell maturation antigen (bcma) single-chain antibody and use thereof
JP6608827B2 (en) Humanized anti-tau (pS422) antibodies and methods of use
US11746153B2 (en) Binding molecules specific for FcγRIIA and uses thereof
CA3000869A1 (en) Antigen-binding polypeptide constructs comprising kappa and lambda light chains and uses thereof
JP2018529733A (en) Humanized anti-human CD19 antibodies and methods of use
EP4031579A2 (en) Anti-klk7 antibodies, anti-klk5 antibodies, multispecific anti-klk5/klk7 antibodies, and methods of use
CA2938931A1 (en) Anti-laminin4 antibodies specific for lg1-3
TW202041535A (en) Antibodies targeting c5ar
WO2022159776A1 (en) Antibodies which bind human fibrin and methods of use
EP4298125A1 (en) Antibodies that bind cd123 and gamma-delta t cell receptors
JP7425725B2 (en) Antibodies, compositions for use in detecting or capturing polypeptides in a sample, and methods for detecting or capturing polypeptides in a sample
WO2022266539A9 (en) ANTIBODIES WHICH BIND HUMAN FIBRIN OR FIBRINOGEN γC DOMAIN AND METHODS OF USE
WO2023245187A2 (en) Antibodies that bind interleukin 13 and methods of use
EP4355773A2 (en) Antibodies which bind human fibrin or fibrinogen gamma c domain and methods of use
WO2024076993A2 (en) Multi-specific antigen binding proteins which bind human fibrin or fibrinogen ?c domain and vascular endothelial growth factor and methods of use
WO2019191668A1 (en) Binding molecules specific for fcyriia and uses thereof
WO2024076990A1 (en) Multi-specific antigen binding proteins which bind human fibrin or fibrinogen ?c domain and vascular endothelial growth factor and methods of use
WO2022262828A1 (en) Anti-il-36r antibody and use thereof
KR20240049265A (en) Antibodies that bind to human fibrin or fibrinogen γC domain and methods of use
TW202409094A (en) Antibodies that bind interleukin 13 and methods of use
WO2023086989A1 (en) Transthyretin (ttr) monomer binding antibodies
AU2020218525A1 (en) Anti-TREM1 antibodies and related methods
TW202334227A (en) Fibrosis treatment with anti-trem2 antibodies
EP4304732A1 (en) Anti-klk7 antibodies, anti-klk5 antibodies, multispecific anti-klk5/klk7 antibodies, and methods of use

Legal Events

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

Ref document number: 22743294

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 22743294

Country of ref document: EP

Kind code of ref document: A1