WO2023133470A2 - Tmprss2 binding antibodies and antigen binding fragments thereof - Google Patents

Tmprss2 binding antibodies and antigen binding fragments thereof Download PDF

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Publication number
WO2023133470A2
WO2023133470A2 PCT/US2023/060179 US2023060179W WO2023133470A2 WO 2023133470 A2 WO2023133470 A2 WO 2023133470A2 US 2023060179 W US2023060179 W US 2023060179W WO 2023133470 A2 WO2023133470 A2 WO 2023133470A2
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Prior art keywords
sequence
seq
antibody
set forth
amino acid
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PCT/US2023/060179
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French (fr)
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WO2023133470A3 (en
Inventor
Anagha DIVEKAR
Susannah KASSMER
Takatoku Oida
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BioLegend, Inc.
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Publication of WO2023133470A2 publication Critical patent/WO2023133470A2/en
Publication of WO2023133470A3 publication Critical patent/WO2023133470A3/en

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
    • G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
    • G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
    • G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
    • G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
    • G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
    • G01N2333/96433—Serine endopeptidases (3.4.21)

Definitions

  • the present disclosure relates, in some aspects, to antibodies or antigen-binding fragments thereof that bind TMPRSS2, as well as methods, systems and kits for detection of TMPRSS2. In certain aspects, the present disclosure relates to antibodies or antigen binding fragments thereof for use in determining levels of TMPRSS2 in a sample containing or suspected of containing TMPRSS2. In some aspects, the present disclosure relates to antibodies or antigen-binding fragments thereof for use in diagnosing or treating an individual with or suspected of having a disease or disorder associated with TMPRSS2.
  • TMPRSS2 transmembrane protease serine 2
  • the transmembrane protease serine 2 (TMPRSS2), is a plasma membrane anchored serine protease that has been associated with both physiological and pathological processes including digestion, tissue remodeling, blood coagulation, fertility, inflammatory responses, tumor cell invasion, apoptosis and pain.
  • the function of TMPRSS2 has also been implicated in facilitating viral pathogenesis for the influenza A virus and human coronaviruses SARS- CoV and SARS-Cov-2.
  • anti-TMPRSS2 antibodies and associated methods are limited in their range of both in vitro and in vivo applications.
  • reagents, devices and methods of detecting TMPRSS2, and diagnosing and/or treating TMPRSS2 related diseases or disorders Provided herein are embodiments that meet such needs.
  • antibodies including antigen-binding fragments thereof, that bind all or a portion thereof of TMPRSS2, compositions containing such antibodies or antigen-binding fragments thereof, combinations of such antibodies or antigen-binding fragments thereof and methods of use.
  • the antibodies or antigenbinding fragments thereof are used in methods of detecting the presence of TMPRSS2 through imaging, including molecular, medical and diagnostic imaging.
  • antibodies or antigen-binding fragments thereof including those that specifically bind to a TMPRSS2, such as a human TMPRSS2, wherein the antibodies or antigen-binding fragments contain particular complementarity determining regions (CDRs), including heavy chain CDRs (i.e., CDRH1, CDRH2, and/or CDRH3) and light chain CDRs (i.e., CDRL1, CDRL2, and/or CDRL3), such as any described herein.
  • CDRs complementarity determining regions
  • the antibody or antigen-binding fragment thereof includes a heavy chain variable domain and a light chain variable domain, such as any described herein.
  • an isolated antibody or antigen binding fragment thereof that binds TMPRSS2 or a portion thereof comprising a) an immunoglobulin heavy chain variable domain comprisingb (i) a heavy chain complementary determining region 1 (CDRH1) comprising the sequence X1X2X3X4X5 (SEQ ID NO: 81), wherein Xi is S, N or D; X2 is Y or S; X3 is G, W or D; X4 is V, M or I; and X5 is S, Q or N; and (ii) a heavy chain complementary determining region 2 (CDRH2) comprising the sequence
  • X1IX2X3X4X5X6X7X8X9X10X11X12X13X14X15X16 (SEQ ID NO: 82), wherein Xi is V, A, Y or W; X2 is W,Y,S or N; X3 is G, P, S or T; X 4 is D, V, G or Y; X 5 is G, D or T; X 6 is S or G; X7 is T, D, R, S or E; Xs is N, T, I or P; X9 is Y, R, K or T; X10 is H, Y or F; Xu is S, T, P, N or A; X12 is A, Q, D or E; X13 is L, K, T or G; X14 is I, F or V; X15 is S or K; and Xi6 is G or no amino acid; and (iii) a heavy chain complementary determining region 3 (CDRH3) comprising the
  • the immunoglobulin heavy chain variable domain includes: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47
  • the immunoglobulin heavy chain variable domain includes: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56; and a CDRH3 comprising the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63.
  • the immunoglobulin light chain variable domain includes: a CDRL1 comprising the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%
  • the immunoglobulin light chain variable domain includes: a CDRL1 comprising a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74; and a CDRL3 comprising a sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 45, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 51, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 57, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 46, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46;
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 52;
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 47, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47;
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53;
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 47
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 53
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 58.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 48, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48;
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 54;
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 48
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 54
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 60.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 45, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45;
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51;
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 61 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 45
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 51
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 61.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 49, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49;
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 55;
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 62 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 49
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 55
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 62.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 50;
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56;
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 50
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 56
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 63.
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 64, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64;
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70;
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 75 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 64
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 70
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 75.
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 65 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:65
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 71 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:71
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 76 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 8
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 66 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 66
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 72 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:72; and the CDRL3 includes the sequence set forth in SEQ ID NO: 77 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%,
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 66
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 72
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 77.
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 67 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 67
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 67
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 73
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 78.
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 79 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 64
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 70
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 79.
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 68 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 68
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 68
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 73
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 78.
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 69 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 69
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 74 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 74
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 80 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 69
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 74
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 80.
  • the CDRH1 includes the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; the CDRH2 includes a sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55, or
  • the CDRH3 includes a sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 51, 52, 53, 54, 55, or 56; the CDRH3 includes a sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:
  • the CDRL1 includes a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69;
  • the CDRL2 includes a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more
  • CDRH1 includes the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50; the CDRH2 includes the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55, or 56; the CDRH3 includes the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63; the CDRL1 includes the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69; the CDRL2 includes the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74; and the CDRL3 includes the sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 57 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 45
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 51
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 57
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 64
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 70
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 75.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 46 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46;
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 52;
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 46
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 52
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 58
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 65
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 71
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 76.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 47 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 59 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 47
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 53
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 59
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 66
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 72
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 77.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 48 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 54
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 8
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 48
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 54
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 60
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 67
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 73
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 78.
  • the CDRH1 includes the sequence set forth in SEQ ID NO:
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51;
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 61 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 45
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 51
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 61
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 64
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 70
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 79.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 49 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 55
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 62 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 49
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 55
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 62
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 68
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 73
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 78.
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 50 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:50
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%,
  • the CDRH1 includes the sequence set forth in SEQ ID NO: 50
  • the CDRH2 includes the sequence set forth in SEQ ID NO: 56
  • the CDRH3 includes the sequence set forth in SEQ ID NO: 63
  • the CDRL1 includes the sequence set forth in SEQ ID NO: 69
  • the CDRL2 includes the sequence set forth in SEQ ID NO: 74
  • the CDRL3 includes the sequence set forth in SEQ ID NO: 80.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 20-26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20-26.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 20-26.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 20 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:20.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 20.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:21.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 21.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 22, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:22.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 22.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 23, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:23.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 23.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 24, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:24.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 24.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:25.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 25.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:26.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 26.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 27-33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27-33.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 27-33.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 27.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 28.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 28.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 9.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 9.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 30.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 30.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 31.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 31.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 32, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 32.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 32.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 33.
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 33.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 20-26 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20-26
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 27-33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27-33.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 20-26, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 27-33.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 20, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 20; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 20, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 27.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 21; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NOS: 28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 21, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 28.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 22, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 22; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 22, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 29.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 23, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 23; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 23, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 30.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 24, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 24; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 24, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 31.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 25; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 32, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO:25
  • the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO:32.
  • the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 26; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO
  • any of the antibody or antigen binding fragment thereof includes one immunoglobulin heavy chain variable domain and one immunoglobulin light chain variable domain. In some embodiments, any of the antibody or antigen binding fragment thereof includes two immunoglobulin heavy chain variable domains and two immunoglobulin light chain variable domains.
  • the antibody or antigen-binding fragment is isolated.
  • the antibody is a humanized antibody, a chimeric antibody or a human antibody.
  • the antibody is a murine antibody.
  • the antibody is an antigen-binding fragment thereof.
  • the antigen-binding fragment thereof is a Fab, Fab'- SH, Fv, scFv, or (Fab')2 fragment.
  • the antibody is a full length or intact antibody.
  • the antibody further comprises a heavy chain constant domain and/or a light chain constant domain.
  • the heavy chain and/or light constant domain is murine or human.
  • the heavy chain constant domain is IgGl, IgG2a, IgG2b or IgM.
  • the antibody is a monoclonal antibody.
  • the antibody is attached to a label.
  • the label is a fluorescent dye, a fluorescent protein, a radioisotope, a chromophores, a metal ion, gold particles, silver particles, magnetic particles, a polypeptides, an enzyme, streptavidin, biotin, a luminescent compound, or an oligonucleotide.
  • the oligonucleotide includes a sample barcode sequence.
  • the oligonucleotide includes a binding site for a primer and an anchor.
  • the detectable marker or label is conjugated directly to the antigen or antigen binding fragment thereof.
  • the detectable marker or label is conjugated to the oligonucleotide.
  • the antibody or antigen binding fragment thereof is non-diffusively immobilized on a solid support.
  • the disclosure provides an isolated antibody that specifically binds to, wherein the isolated antibody competes binding to the with an antibody described herein.
  • the antibody is a monoclonal antibody. In certain embodiments, the antibody is a humanized antibody. In certain embodiments, the antibody comprises one or more human framework regions.
  • a combination of antibodies or antigen-binding fragments thereof wherein the combination comprises two or more anti-TMPRSS2 antibodies or antigen-binding fragments described herein.
  • the two or more antibodies or antigen-binding fragments comprise one or more first antibody or antigenbinding fragment thereof that binds to a first epitope or region within TMPRSS2; and one or more second antibody or antigen-binding fragment thereof that binds to a second epitope or region within TMPRSS2.
  • the one or more first antibody or antigen-binding fragments thereof, and the one or more second antibody or antigen-binding fragments thereof bind to a non-overlapping epitope or region of TMPRSS2 (e.g., human TMPRSS2) and/or do not compete for binding to TMPRSS2.
  • the antibody is conjugated to a detectable marker or label.
  • the at least one of the antibodies or antigen-binding fragments of the combination of two or more anti- TMPRSS2 antibodies or antigen-binding fragments described herein, optionally the one or more first antibody or antigen-binding fragment thereof or the one or more second antibody or antigen-binding fragment thereof is conjugated to a label.
  • the at least one of the antibodies or antigen-binding fragments is attached or immobilized to a solid support.
  • the one or more first or second antibody or antigen-binding fragment is attached or immobilized to a solid support and the other of the one or more first or second antibody or antigen-binding fragment is conjugated to a label.
  • the label is a fluorescent dye, a fluorescent protein, a radioisotope, a chromophore, a metal ion, gold particles, silver particles, magnetic particles, a polypeptide, an enzyme, streptavidin, biotin, a luminescent compound, or an oligonucleotide.
  • the solid support is a bead, a column, an array, an assay plate, a microwell, a stick, a filter, or a strip.
  • the antibody is non- diffusively immobilized on a solid support.
  • the device is a rapid detection device or a rapid diagnostic device.
  • the disclosure features an isolated nucleic acid encoding an isolated antibody described herein.
  • the disclosure also provides an expression vector comprising the nucleic acid described herein. Further, the disclosure also provides an isolated host cell comprising the expression vector described herein.
  • the antibody or antigen binding fragment thereof provided herein can be used in the detection of TMPRSS2 in a sample.
  • the antibody or antigen binding fragment thereof binds to a cell expressing TMPRSS2 in a sample.
  • the sample includes immune cells.
  • the sample includes a heterogenous population of immune cells.
  • the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs).
  • the sample includes a cell with a disease or disorder.
  • the disease or disorder is a cancer, an autoimmune disorder, an inflammatory disorder, a neurologic disorder, or an infection.
  • the cancer is acute myeloid leukemia, acute lymphoblastic leukemia, colorectal, ovarian, gynecologic, liver, glioblastoma, Hodgkin lymphoma, chronic lymphocytic leukemia, esophagus, gastric, pancreas, colon, kidney, head and neck, lung and melanoma.
  • the detection includes the use of a single antibody or antigen binding fragment thereof to bind a portion of TMPRSS2. In some embodiments, the detection includes the use of two antibody or antigen binding fragments thereof, each capable of binding to a different portion of TMPRSS2.
  • the detection of TMPRSS2 is on the surface of a cell. In some embodiments, the detection of TMPRSS2 is intracellular. In some embodiments, the detection of TMPRSS2 indicates the presence or absence of a disease or disorder. In some embodiments, the detection is performed in vitro. In some embodiments, the detection is performed in vivo. [0062] In some embodiments, the antibody or antigen binding fragment thereof binds to a TMPRSS2 expressing cell. In some embodiments, the binding to the TMPRSS2 expressing cell decreases the production of androgenic hormones. In some embodiments, the binding to the TMPRSS2 expressing cell inhibits proteolytic cleavage of ACE2 receptor.
  • the binding to the TMPRSS2 expressing cell inhibits or reduces cleavage of coronavirus spike glycoproteins. In some embodiments, the binding to the TMPRSS2 expressing cell inhibits or reduces viral uptake into a host cell.
  • kits comprising the antibody or antigen binding fragment thereof of any one of embodiments described herein.
  • the kit is a diagnostics kit configured to detect TMPRSS2 in a biological sample.
  • composition comprising the antibody or antigen binding fragment thereof of any of the embodiments described herein and a pharmaceutically acceptable excipient.
  • the antibody or antigen binding fragment thereof of is used as an adjuvant or in conjunction with an adjuvant.
  • an isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of the agent of any of embodiments described herein.
  • the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in any of SEQ ID NOs: 6-12.
  • the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 6.
  • the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 7.
  • the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 8.
  • the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 9. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 10. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 11. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 12.
  • an isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin light chain variable domain of the agent of any of the embodiments described herein.
  • the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in any of SEQ ID NOs: 13-19.
  • the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 13.
  • the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 14.
  • the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 15.
  • the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 16. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 17. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 18. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 19.
  • nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain and the immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any of the embodiments described herein.
  • nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in any of SEQ ID NOs: 6-12; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in any of SEQ ID NOs: 13-19.
  • the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 6; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 13.
  • the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 7; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 14.
  • the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 8; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 15.
  • a recombinant expression vector comprising the isolated nucleic acid of any of the embodiments described herein.
  • a recombinant expression vector comprising a first expression cassette and a second expression cassette, wherein the first expression cassette includes a nucleic acid molecule comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of any one of the embodiments described herein and the second expression cassette includes a nucleic acid molecule comprising a nucleotide sequence that encodes an immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any one of any of the embodiments described herein.
  • a recombinant expression vector comprising a first expression cassette and a second expression cassette, wherein the first expression cassette includes a nucleic acid molecule comprising the nucleotide sequence of any of the embodiments described herein, and the second expression cassette includes a nucleic acid molecule comprising the nucleotide sequence of any of the embodiments described herein.
  • the first and second expression cassettes include a promoter.
  • an agent-drug conjugate comprising antibody or antigen binding fragment thereof of any of the embodiments described herein.
  • a composition comprising the antibody-drug conjugate and a pharmaceutically acceptable carrier.
  • a method of detecting TMPRSS2 comprising a) contacting a sample with the antibody or antigen binding fragment thereof of any of the embodiments described herein under conditions to bind said antibody or antigen binding fragment thereof to a TMPRSS2 receptor on said sample, wherein the binding generates the production of a receptor/antibody or antigen binding fragment thereof of complex; b) detecting the presence of the receptor/antibody or antigen binding fragment thereof of complexes; c) wherein the detecting includes the presence or absence of the TMPRSS2 receptor on said sample.
  • a method of treating or preventing a disease or disorder associated with TMPRSS2 in a subject comprising: a) contacting a sample known or suspected to contain TMPRSS2 with the antibody or antigen binding fragment thereof of any of the embodiments described herein b) detecting the presence of complexes comprising TMPRSS2 and the antibody or antigen binding fragment thereof; wherein the presence of the complexes indicates the presence of a disease or disorder; and c) administering to the subject the antibody or antigen binding fragment thereof of any of any of the embodiments described herein.
  • a method of diagnosing a disease or disorder comprising: a) isolating a sample from a subject b) incubating the sample with the antibody or antigen binding fragment thereof of any of any of the embodiments described herein, for a period of time sufficient to generate TMPRSS2:anti-TMPRSS2 complexes; c) detecting the presence or absence of the TMPRSS2:anti-TMPRSS2 complexes from the isolated tissue, and d) associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
  • the increase of TMPRSS2 over a control level in the location of interest of the tissue sample is indicative of a disease or disorder in a subject.
  • the method is performed in vitro. In some embodiments, the method is performed in vivo. In some embodiments, the detection includes intracellular detection. In some embodiments, the detection includes detection on the surface of a cell. In some embodiments, the detection includes hybridization of a detectable moiety to the antibody or antigen binding fragment thereof. In some embodiments, the sample is contacted with a second antibody. In some embodiments, the second antibody is an antibody comprising a detectable moiety. In some embodiments, the detectable moiety includes an oligonucleotide. In some embodiments, the detectable moiety includes a fluorescent label. In some embodiments, the measurement includes sequencing. In some embodiments, the detectable moiety includes immunofluorescence. In some embodiments, the sample is a formalin-fixed paraffin-embedded sample. In some embodiments, the sample includes a cell. In some embodiments, the sample includes a tissue sample.
  • the sample includes immune cells.
  • the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs).
  • the sample includes a tissue or cells associated with a disease or disorder.
  • the disease or disorder is a cancer, an autoimmune disorder, an inflammatory disorder, or an infection.
  • the disease or disorder is chosen from non-viral cancers, virus-associated cancers, cancers associated with HBV infection, cancers associated with Epstein-Barr virus (EBV) infection, cancers associated with polyomavirus infection, erythema nodosum leprosum (ENL), autoimmune diseases, autoimmune inflammation, autoimmune thyroid diseases, B-cell lymphoma, T-cell lymphoma, acute myeloid leukemia, Hodgkin's Disease, acute myelogenous leukemia, acute myelomonocytic leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, B cell large cell lymphoma, malignant lymphoma, acute leukemia, lymphosarcoma cell leukemia, B-cell leukemias, myelodysplastic syndromes, solid phase cancer, herpes viral infections, and/or rejection of transplanted tissues or organs.
  • EBV Epstein-Barr virus
  • the antibody or antigen binding fragment thereof can be used in a method of associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
  • the antibody or antigen binding fragment thereof can be used in a method of detecting TMPRSS2 in a tissue sample.
  • the method includes generating a nucleic acid molecule comprising all or a portion of the sequence of the oligonucleotide or a complement thereof.
  • the antibody or antigen binding fragment thereof can be used in the construction of a protein library.
  • the construction of a protein library includes sequencing.
  • the construction of a protein library includes the use of flow cytometry.
  • FIGs. 1A-1B show the amino acid sequences of the variable domains of the immunoglobulin heavy (SEQ ID NOS:20-26 and light (SEQ ID NOS: 27-33) chains of the seven different anti-TMPRSS2 antibodies (AB 1-7) provided herein.
  • the CDR regions of each of the heavy and light chains are shown in bold and underline.
  • FIGs. 2A-2E depict staining profiles of exemplary tested anti-TMPRSS2 antibodies AB1, AB2, AB4, and AB6 (FIGs. 2A-2D) compared to a commercially available antibody (FIG. 2E) on Caco-2 cells, assessed by flow cytometry.
  • FIGs. 3A-3F depict staining profiles of exemplary tested anti-TMPRSS2 antibodies
  • FIGs. 4A-4C depict staining profiles of exemplary tested anti-TMPRSS2 antibodies AB1, AB2, AB4, and AB6 compared to control on lymphocytes (FIG. 4A), monocytes (FIG. 4B), or granulocytes (FIG. 4C), assessed by flow cytometry.
  • FIGs. 5A-5B depict staining of formalin-fixed paraffin-embedded (FFPE) samples with exemplary anti-human TMPRSS2 antibodies AB1, AB2, AB3, AB5 or AB7 on human kidney paraffin sections (FIG. 5 A) and non-TMPRSS2 expressing lymph node sections (FIG. 5B), assessed by immunohistochemistry.
  • FFPE formalin-fixed paraffin-embedded
  • FIGs. 6A-6D depict assessment of the functional activity of exemplary anti-human TMPRSS2 antibody AB1.
  • the effect of AB1 on TMPRSS2 protease activity was compared to Nafamostat or control in both platelet-rich plasma (FIG. 6A) and using recombinant TMPRSS2 (FIG. 6B).
  • the effect of AB1 on TMPRSS2 induced cell migration was compared to Nafamostat or control in CaCo-2 cells (FIG. 6C).
  • the effect of AB1 on TMPRSS2 induced cell migration was compared to Nafamostat or four separate commercially available antibodies in CaCo-2 cells (FIG. 6D).
  • antibodies that bind TMPRSS2 including antigen-binding fragments thereof, nucleic acids encoding such antibodies and antigen-binding fragments, and cells, such as recombinant cells for expressing and production of these antibodies and antigenbinding fragments that can bind to under physiological and/or in vitro conditions.
  • Also provided are methods of producing and using the antibodies and antigen-binding fragments such as in methods for detecting TMPRSS2 in a sample from an individual, including methods for laboratory/ research purposes (e.g., flow cytometry, ELISA, and/or Western blot), and/or for the use and treatment and/or prevention of various diseases or disorders through the delivery of pharmaceutical or other compositions that contain such antibodies or antigen-binding fragments thereof.
  • methods for laboratory/ research purposes e.g., flow cytometry, ELISA, and/or Western blot
  • methods for producing and using the antibodies and antigen-binding fragments such as in methods for detecting TMPRSS2 in a sample from an individual, including methods for laboratory/ research purposes (e.g., flow cytometry, ELISA, and/or Western blot), and/or for the use and treatment and/or prevention of various diseases or disorders through the delivery of pharmaceutical or other compositions that contain such antibodies or antigen-binding fragments thereof.
  • antibody includes antigen binding fragments thereof that retain binding specificity. For example, there are a number of well characterized antigen binding fragments.
  • pepsin digests an antibody C-terminal to the disulfide linkages in the hinge region to produce F(ab)'2, a dimer of Fab which itself is a light chain joined to VH-CH1 by a disulfide bond.
  • the F(ab)'2 may be reduced under mild conditions to break the disulfide linkage in the hinge region thereby converting the (Fab')2 dimer into an Fab' monomer.
  • the Fab' monomer is essentially an Fab with part of the hinge region (see, Fundamental Immunology, W.E.
  • antigen binding fragments are defined in terms of the digestion of an intact antibody, one of skill will appreciate that fragments can be synthesized de novo either chemically or by utilizing recombinant DNA methodology.
  • antibody as used herein also includes antigen binding fragments either produced by the modification of whole antibodies or synthesized using recombinant DNA methodologies.
  • An antibody as described herein can consist of one or more polypeptides substantially encoded by immunoglobulin genes or fragments of immunoglobulin genes.
  • the recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as myriad immunoglobulin variable region genes.
  • Light chains are classified as either kappa or lambda.
  • Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively.
  • the antibody is IgG (e.g., IgGl, IgG2, IgG3, IgG4), IgM, IgA, IgD, or IgE.
  • a typical immunoglobulin (antibody) structural unit is known to comprise a tetramer.
  • Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light” (about 25 kD) and one "heavy” chain (about 50-70 kD).
  • the N-terminus 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 chains respectively.
  • substitution variants have at least one amino acid residue removed and a different residue inserted in its place.
  • the sites of greatest interest for substitutional mutagenesis include the hypervariable regions, but framework alterations are also contemplated. Examples of conservative substitutions are described above.
  • Substantial modifications in the biological properties of the antibody are accomplished by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a P-sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain.
  • Naturally occurring residues are divided into groups based on common side-chain properties:
  • Non-conservative substitutions are made by exchanging a member of one of these classes for another class.
  • One type of substitution that can be made is to change one or more cysteines in the antibody, which may be chemically reactive, to another residue, such as, without limitation, alanine or serine.
  • the substitution can be made in a CDR or framework region of a variable domain or in the constant region of an antibody.
  • the cysteine is canonical (e.g., involved in disulfide bond formation).
  • cysteine residues not involved in maintaining the proper conformation of the antibody also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant cross-linking.
  • cysteine bond(s) may be added to the antibody to improve its stability, particularly where the antibody is an antigen binding fragment such as an Fv fragment.
  • Antibodies include VH-VL dimers, including single chain antibodies (antibodies that exist as a single polypeptide chain), such as single chain Fv antibodies (sFv or scFv) in which a variable heavy and a variable light domains are joined together (directly or through a peptide linker) to form a continuous polypeptide.
  • the single chain Fv antibody is a covalently linked VH-VL which may be expressed from a nucleic acid including VH- and VL- encoding sequences either joined directly or joined by a peptide-encoding linker (e.g., Huston, et al. Proc. Nat. Acad. Sci. USA, 85:5879-5883, 1988).
  • the VH and VL domains associate non-covalently.
  • the antibody can be another fragment. Other fragments can also be generated, e.g., using recombinant techniques, as soluble proteins or as fragments obtained from display methods.
  • Antibodies can also include diantibodies and miniantibodies.
  • Antibodies of the disclosure also include heavy chain dimers, such as antibodies from camelids.
  • an antibody is dimeric.
  • the antibody may be in a monomeric form that has an active isotype.
  • the antibody is in a multivalent form, e.g., a trivalent or tetravalent form.
  • an “antibody fragment” or “antigen binging fragment thereof’ comprises a portion of an intact antibody, the antigen binding and/or the variable region of the intact antibody.
  • Atibody fragments or antigen binding fragments thereof include but are not limited to Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, disulfide-linked Fvs (dsFv), Fd fragments, Fd' fragments; diabodies; linear antibodies (see U.S. Pat. No.5, 641, 870, Example 2; Zapata et al, Protein Eng.
  • single-chain antibody molecules including single-chain Fvs (scFv) or single-chain Fabs (scFab); antigen-binding fragments of any of the above and multispecific antibodies from from antibody fragments.
  • scFv single-chain Fvs
  • scFab single-chain Fabs
  • Fv is composed of one heavy- and one light-chain variable region domain linked by non-covalent association. From the folding of these two domains emanate six complementarity determining regions (CDR) (3 in each from the heavy and light chain) that contribute the amino acid residues for antigen binding and confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although, in some cases, at a lower affinity than the entire binding site.
  • CDR complementarity determining regions
  • dsFv refers to an Fv with an engineered intermolecular disulfide bond, which stabilizes the VH-VL pair.
  • An “Fd fragment” is a fragment of an antibody containing a variable domain (VH) and one constant region domain (CHI) of an antibody heavy chain.
  • a "Fab fragment” is an antibody fragment that results from digestion of a full-length immunoglobulin with papain, or a fragment having the same structure that is produced synthetically, e.g., by recombinant methods.
  • a Fab fragment contains a light chain (containing a VL and CL) and another chain containing a variable domain of a heavy chain (VH) and one constant region domain of the heavy chain (CHI).
  • a "F(ab')2 fragment” is an antibody fragment that results from digestion of an immunoglobulin with pepsin at pH 4.0-4.5, or a fragment having the same structure that is produced synthetically, e.g., by recombinant methods.
  • the F(ab')2 fragment essentially contains two Fab fragments where each heavy chain portion contains an additional few amino acids, including cysteine residues that form disulfide linkages joining the two fragments.
  • a "Fab 1 fragment” is a fragment containing one half (one heavy chain and one light chain) of the F(ab')2 fragment.
  • An "Fd 1 fragment” is a fragment of an antibody containing one heavy chain portion of a F(ab')2 fragment.
  • An "Fv 1 fragment” is a fragment containing only the VH and VL domains of an antibody molecule.
  • an "scFv fragment” refers to an antibody fragment that contains a variable light chain (VL) and variable heavy chain (VH), covalently connected by a polypeptide linker in any order.
  • the linker is of a length such that the two variable domains are bridged without substantial interference.
  • Exemplary linkers are (Gly-Ser)n residues with some Glu or Lys residues dispersed throughout to increase solubility.
  • Diabodies are dimeric scFv; diabodies typically have shorter peptide linkers than scFvs, and preferentially dimerize.
  • variable region and “variable domain” refer to the portions of the light and heavy chains of an antibody that include amino acid sequences of complementary determining regions (CDRs, e.g., HCDR1, HCDR2, HCR3, LCDR1, LCDR2, and LCDR3) and framework regions (FRs).
  • CDRs complementary determining regions
  • FRs framework regions
  • the variable domain for the heavy and light chains is commonly designated VH and VL, respectively.
  • the variable domain is included on Fab, F(ab’)2, Fv and scFv antigen binding fragments described herein, and is involved in specific antigen recognition.
  • CDR complementarity-determining region
  • the sequences of the framework regions of different light or heavy chains are relatively conserved within a species.
  • the framework region of an antibody that is the combined framework regions of the constituent light and heavy chains, serves to position and align the CDRs in three-dimensional space.
  • the amino acid sequences of the CDRs and framework regions can be determined using various well-known definitions in the art, e.g., Kabat, North method (see, e.g., North et al., J Mol Biol. 406(2):228-256, 2011), Chothia, international ImMunoGeneTics database (IMGT), and AbM (see, e.g., Johnson etal, supra, Chothia & Lesk, 1987, Canonical structures for the hypervariable regions of immunoglobulins. J. Mol. Biol. 196, 901-917; Chothia C. et al., 1989, Conformations of immunoglobulin hypervariable regions. Nature 342, 877-883; Chothia C.
  • chimeric antibody refers to an immunoglobulin molecule in which (a) the constant region, or a portion thereof, is altered, replaced or exchanged so that the antigen binding site (variable region) is linked to a constant region of a different or altered class, effector function and/or species, or an entirely different molecule which confers new properties to the chimeric antibody, e.g., an enzyme, toxin, hormone, growth factor, drug, etc.; or (b) the variable region, or a portion thereof, is altered, replaced or exchanged with a variable region, or portion thereof, having a different or altered antigen specificity; or with corresponding sequences from another species or from another antibody class or subclass.
  • humanized antibody refers to an immunoglobulin molecule in CDRs from a donor antibody are grafted onto human framework sequences. Humanized antibodies may also comprise residues of donor origin in the framework sequences. The humanized antibody can also comprise at least a portion of a human immunoglobulin constant region. Humanized antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences.
  • Humanization can be performed using methods known in the art (e.g., Jones et al., Nature 321 :522-525; 1986; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen etal., Science 239: 1534-1536, 1988); Presta, Curr. Op. Struct. Biol. 2:593-596, 1992; U.S. Patent No. 4,816,567), including techniques such as “superhumanizing” antibodies (Tan et al., J. Immunol. 169: 1119, 2002) and "resurfacing” (e.g., Staelens et al., Mol. Immunol. 43: 1243, 2006; and Roguska et al., Proc. Natl. Acad. Sci USA 91 : 969, 1994).
  • methods known in the art e.g., Jones et al., Nature 321 :522-525; 1986; Riechmann et al.,
  • recombinant when used with reference, e.g., to a cell, or nucleic acid, protein, or vector, indicates that the cell, nucleic acid, protein or vector, has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified.
  • recombinant cells express genes that are not found within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all.
  • antigen e.g., a polypeptide, polynucleotide, carbohydrate, lipid, chemical moiety, or combinations thereof (e.g., phosphorylated or glycosylated polypeptides, etc.).
  • a polypeptide, polynucleotide, carbohydrate, lipid, chemical moiety, or combinations thereof e.g., phosphorylated or glycosylated polypeptides, etc.
  • Antibodies bind to an “epitope” on an antigen.
  • the epitope is the localized site on the antigen that is recognized and bound by the antibody.
  • Epitopes can include a few amino acids or portions of a few amino acids, e.g., 5 or 6, or more, e.g., 20 or more amino acids, or portions of those amino acids.
  • the epitope includes non-protein components, e.g., from a carbohydrate, nucleic acid, or lipid. In some cases, the epitope is a three- dimensional moiety.
  • the epitope can be comprised of consecutive amino acids, or amino acids from different parts of the protein that are brought into proximity by protein folding (e.g., a discontinuous epitope).
  • a discontinuous epitope e.g., a discontinuous epitope.
  • An epitope typically includes at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial conformation.
  • Methods of determining spatial conformation of epitopes include, for example, x-ray crystallography and 2-dimensional nuclear magnetic resonance. See, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, Glenn E. Morris, Ed (1996).
  • a “label” or a “detectable moiety” is a diagnostic agent or component detectable by spectroscopic, radiological, photochemical, biochemical, immunochemical, chemical, or other physical means.
  • exemplary labels include radiolabels (e.g., in In, " m Tc, 131 I, 67 Ga) and other FDA-approved imaging agents. Additional labels include 32 P, fluorescent dyes, electron-dense reagents, enzymes, biotin, digoxigenin, or haptens and proteins or other entities which can be made detectable, e.g., by incorporating a radiolabel into the targeting agent. Any method known in the art for conjugating a nucleic acid or nanocarrier to the label may be employed, e.g., using methods described in Hermanson, Bioconjugate Techniques 1996, Academic Press, Inc., San Diego.
  • a “labeled” or “tagged” antibody or agent is one that is bound, either covalently, through a linker or a chemical bond, or noncovalently, through ionic, van der Waals, electrostatic, or hydrogen bonds to a label such that the presence of the antibody or agent may be detected by detecting the presence of the label bound to the antibody or agent.
  • the terms “specific for,” “specifically binds,” and like terms refer to a molecule (e.g., antibody or antigen binding fragment) that binds to a target with at least 2-fold greater affinity than non-target compounds, e.g., at least any of 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 25-fold, 50-fold, or 100-fold greater affinity.
  • a target e.g., TMPRSS2
  • TMPRSS2 will typically bind the target with at least a 2-fold greater affinity than a non-target.
  • Specificity can be determined using standard methods, e.g., solid-phase ELISA immunoassays (see, e.g., Harlow & Lane, Using Antibodies, A Laboratory Manual (1998) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity).
  • an antibody target e.g., antigen, analyte, immune complex
  • an antibody that binds a given antibody target typically binds to at least 2/3 of the antibody targets in a solution (e.g., at least any of 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100%).
  • a solution e.g., at least any of 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100%.
  • a “control” sample or value refers to a sample that serves as a reference, usually a known reference, for comparison to a test sample.
  • a test sample can be taken from a test condition, e.g., in the presence of a test compound, and compared to samples from known conditions, e.g., in the absence of the test compound (negative control), or in the presence of a known compound (positive control).
  • a control can also represent an average value or a range gathered from a number of tests or results.
  • controls can be designed for assessment of any number of parameters.
  • a control can be devised to compare therapeutic benefit based on pharmacological data (e.g., half-life) or therapeutic measures (e.g., comparison of benefit and/or side effects).
  • Controls can be designed for in vitro applications.
  • One of skill in the art will understand which controls are valuable in a given situation and be able to analyze data based on comparisons to control values. Controls are also valuable for determining the significance of data. For example, if values for a given parameter are widely variant in controls, variation in test samples will not be considered as significant.
  • nucleic acids or polypeptide sequences refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region, when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection (see, e.g., NCBI web site ncbi.nlm.nih.gov/BLAST/ or the like).
  • identity exists over a region that is at least about 25 amino acids or nucleotides in length, or more preferably over a region that is 50-100 or more amino acids or nucleotides in length.
  • sequence comparison typically one sequence acts as a reference sequence, to which test sequences are compared.
  • test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated.
  • sequence algorithm program parameters Preferably, default program parameters can be used, or alternative parameters can be designated.
  • sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.
  • a “comparison window”, as used herein, includes reference to a segment of any one of the number of contiguous positions selected from the group consisting of from 20 to 600, usually about 50 to about 200, more usually about 100 to about 150 in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are well- known in the art.
  • BLAST and BLAST 2.0 are used, with the parameters described herein, to determine percent sequence identity for the nucleic acids and proteins of the disclosure.
  • Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/).
  • This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence.
  • T is referred to as the neighborhood word score threshold (Altschul et al., supra).
  • a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negativescoring residue alignments; or the end of either sequence is reached.
  • the BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment.
  • nucleic acid refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or double-stranded form, and complements thereof.
  • the term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides.
  • Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).
  • nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences, as well as the sequence explicitly indicated.
  • degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer c/ al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); Rossolini etal., Mol. Cell. Probes 8:91-98 (1994)).
  • polypeptide refers to a polymer of amino acid residues.
  • the terms encompass to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non- naturally occurring amino acid polymer.
  • amino acid refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids.
  • Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, y- carboxyglutamate, and O-phosphoserine.
  • Amino acid analogs refers to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid.
  • Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid.
  • Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes.
  • the term “compete”, as used herein with regard to an antibody means that a first antibody, or an antigen-binding portion thereof, competes for binding with a second antibody, or an antigen-binding portion thereof, where binding of the first antibody with its cognate epitope is detectably decreased in the presence of the second antibody compared to the binding of the first antibody in the absence of the second antibody.
  • the alternative, where the binding of the second antibody to its epitope is also detectably decreased in the presence of the first antibody can, but need not be the case. That is, a first antibody can inhibit the binding of a second antibody to its epitope without that second antibody inhibiting the binding of the first antibody to its respective epitope.
  • each antibody detectably inhibits the binding of the other antibody with its cognate epitope or ligand, whether to the same, greater, or lesser extent, the antibodies are said to “cross-compete” with each other for binding of their respective epitope(s).
  • Both competing and cross-competing antibodies are encompassed by the present disclosure. Regardless of the mechanism by which such competition or crosscompetition occurs (e.g., steric hindrance, conformational change, or binding to a common epitope, or portion thereof, and the like), the skilled artisan would appreciate, based upon the teachings provided herein, that such competing and/or cross-competing antibodies are encompassed and can be useful for the methods disclosed herein.
  • RIA solid phase direct or indirect radioimmunoassay
  • EIA solid phase direct or indirect enzyme immunoassay
  • sandwich competition assay see Stahli et al., Methods in Enzymology 9:242-253 (1983)
  • solid phase direct biotin-avidin EIA see Kirkland et al., J. Immunol. 137:3614-3619 (1986)
  • solid phase direct labeled assay solid phase direct labeled sandwich assay
  • solid phase direct labeled sandwich assay see Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Press (1988)
  • solid phase direct label RIA using 1-125 label see Morel et al., Molec. Immunol.
  • 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. Usually, when a competing antibody is present in excess, it will inhibit specific binding of a reference antibody to a common antigen by at least 50 or 75%.
  • TMPRSS2 refers to human TMPRSS2 proteins, isoforms or variants thereof, including naturally occurring variants of human TMPRSS2, such as splice variants or allelic variants.
  • the amino acid sequence of an exemplary human TMPRSS2 is shown in SEQ ID NO: 1.
  • the amino acid sequence of another exemplary human TMPRSS2 is shown in SEQ ID NO: 2.
  • human TMPRSS2 can refer to a variant, such as an allelic variant or splice variant, that exhibits at least or at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any of SEQ ID NOs: 1-2.
  • the provided antibodies or antigen-binding fragments may exhibit cross-reactive binding to another mammalian TMPRSS2 protein, such as murine TMPRSS2, or a primate TMPRSS2.
  • TMPRSS2 isoform 1 amino acid sequence (SEQ ID NO:1; UniProt 015393-1) MALNSGSPPAIGPYYENHGYQPENPYPAQPTVVPTVYEVHPAQYYPSPVPQYAPRVL TQASNPVVCTQPKSPSGTVCTSKTKKALCITLTLGTFLVGAALAAGLLWKFMGSKCS NSGIECDSSGTCINPSNWCDGVSHCPGGEDENRCVRLYGPNFILQVYSSQRKSWHPV CQDDWNENYGRAACRDMGYKNNFYSSQGIVDDSGSTSFMKLNTSAGNVDIYKKLY HSDACSSKAVVSLRCIACGVNLNSSRQSRIVGGESALPGAWPWQVSLHVQNVHVCG GSIITPEWIVTAAHCVEKPLNNPWHWTAFAGILRQSFMFYGAGYQVEKVISHPNYDS KTKNNDIALMKLQKPLTFNDLVKPVCLPNPGMMLQP
  • TMPRSS2 isoform 2 amino acid sequence (SEQ ID NO:2; UniProt 015393-2)
  • any of the exemplary human TMPRSS2 amino acid sequences comprises a cytoplasmic domain.
  • the cytoplasmic domain comprises the sequence set forth in the SEQ ID NO: 3.
  • any of the exemplary human TMPRSS2 amino acid sequences comprises a transmembrane domain.
  • the transmembrane domain comprises the sequence set forth in the SEQ ID NO: 4.
  • any of the exemplary human TMPRSS2 amino acid sequences comprises an extracellular domain.
  • the extracellular domain comprises the sequence set forth in the SEQ ID NO: 5.
  • solid support is meant a non-aqueous matrix to which an antibody according to the provided disclosure can adhere or attach.
  • solid supports include, but are not limited to, a microtiter plate, a membrane (e.g. , nitrocellulose), a bead, a dipstick, a thin-layer chromatographic plate, or other solid medium.
  • an "individual” or a “subject” is a mammal.
  • a “mammal” for purposes of treatment includes humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, horses, rabbits, cattle, pigs, hamsters, gerbils, mice, ferrets, rats, cats, etc.
  • the individual or subject is human.
  • antibodies including antigen binding fragments thereof, that specifically bind to TMPRSS2.
  • the provided antibodies include monoclonal antibodies and antigen-binding fragments thereof that bind TMPRSS2 that provide superior target specificity, signal-to-noise ratios, and the like as compared to other reported antibodies.
  • methods for producing anti-TMPRSS2 antibodies, and methods for detecting and using such antibodies are also provided herein.
  • Transmembrane serine protease 2 (TMPRSS2) is s type II transmembrane protein containing approximately 492 amino acids encoded by the TMPRSS2 gene.
  • the TMPRSS2 gene is widely conserved and has two isoforms, which differ only in the N-terminal, cytoplasmic tail, with isoform 1 containing 37 additional amino acids on the tail which are not present in isoform 2. Both TMPRSS2 isoforms are autocatalytically activated from the inactive zymogen form.
  • TMPRSS2 has been associated with physiological and pathological processes such as digestion, tissue remodeling, blood coagulation, fertility, inflammatory responses, tumor cell invasion, apoptosis and pain, and is upregulated by androgenic hormones in prostate cancer cells.
  • TMPRSS2 activates several substrates that include pro-hepatocyte growth factor/HGF, the protease activated receptor-2/F2RLl or matriptase/ST14 leading to extracellular matrix disruption and metastasis of prostate cancer cells.
  • TMPRSS2 has been suggested to facilitate human coronaviruse SARS-CoV and SARS-CoV-2 infections via two independent mechanisms: proteolytic cleavage of ACE2 receptor which promotes viral uptake, and cleavage of coronavirus spike glycoproteins which activates the glycoprotein for host cell entry.
  • TMPRSS2 has been shown to activate the spike glycoproteins of human coronavirus 229E (HCoV-229E) and human coronavirus EMC (HCoV-EMC) and the fusion glycoproteins F0 of Sendai virus (SeV), human metapneumovirus (HMPV), human parainfluenza 1, 2, 3, 4a and 4b viruses (HPIV).
  • SARS-CoV and SARS-CoV-2 use angiotensinconverting enzyme 2 (ACE2) and TMPRSS2 to facilitate entry to cells, but with SARS-CoV-2 human-to-human transmission is much higher than SARS-CoV (Thunders et al. J Clin Pathol 73(12):773-776 (2020); Lucas et al. Cancer Discovery 4:1310-1325 (2014); Wilson et al. Biochem J. 388-967-972 (2005); Ko et al Cancer Res. 75:2949-2960 (2015)).
  • ACE2 angiotensinconverting enzyme 2
  • TMPRSS2 TMPRSS2
  • any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of TMPRSS2. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the extracellular domain of TMPRSS2. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the extracellular domain of TMPRSS2 set forth in SEQ ID NO: 5. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the transmembrane domain of TMPRSS2.
  • any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the transmembrane domain of TMPRSS2 set forth in SEQ ID NO: 4. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the cytoplasmic domain of TMPRSS2. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the cytoplasmic domain of TMPRSS2 set forth in SEQ ID NO: 3.
  • any of the antibodies or antigen binding fragments thereof is a TMPRSS2 antibody or antigen binding fragment thereof.
  • the antibody or antigen binding fragment thereof is isolated (e.g., separated from a component of its natural environment (e.g., an animal, a biological sample)).
  • the anti- antibody is a humanized antibody, or an antigen binding fragment thereof.
  • the antibody is a derivative of a humanized antibody that binds.
  • the antibody binds under laboratory conditions (e.g., binds in vitro, binds in a flow cytometry assay, binds in an ELISA).
  • the antibody binds under physiological conditions (e.g., binds in a cell in a subject).
  • the antibodies provided herein comprise at least one immunoglobulin heavy chain variable domain and at least one immunoglobulin light chain variable domain.
  • an antibody described herein comprises two immunoglobulin heavy chain variable domains and two immunoglobulin light chain variable domains.
  • each immunoglobulin heavy chain variable domain of the antibody comprises first, second, and third heavy chain complementarity determining regions (CDRs; HCDR1, HCDR2, and HCDR3)
  • each immunoglobulin light chain variable domain of the antibody comprises first, second, and third light chain CDRs (LCDR1, LCDR2, and LCDR3).
  • the antibodies are antigen binding fragments such as Fab, F(ab’)2, Fv or scFv.
  • the antigen binding fragments can be generated using any means known in the art including, chemical digestion (e.g., papain or pepsin) and recombinant methods. Methods for isolating and preparing recombinant nucleic acids are known to those skilled in the art (see, Sambrook et al., Molecular Cloning. A Laboratory Manual (2d ed. 1989); Ausubel et al., Current Protocols in Molecular Biology (1995)).
  • the antibodies can be expressed in a variety of host cells, including E. coli, other bacterial hosts, yeast, and various higher eukaryotic cells such as the COS, CHO, and HeLa cells lines and myeloma cell lines.
  • the disclosure provides an isolated antibody or antigen binding fragment thereof that specifically binds to CD56, wherein the isolated antibody comprises: a) an immunoglobulin heavy chain variable domain comprising: (i) a heavy chain complementary determining region 1 (CDRH1) comprising the sequence comprising the sequence X1X2X3X4X5 (SEQ ID NO: 81), wherein Xi is S, N or D; X2 is Y or S; X3 is G, W or D; X 4 is V, M or I; and X5 is S, Q or N; (ii) a heavy chain complementary determining region 2 (CDRH2) comprising the sequence X1IX2X3X4X5X6X7X8X9X10X11X12X13X14X15X16 (SEQ ID NO: 82), wherein Xi is V, A, Y or W; X2 is W,Y,S or N; X3 is G, P, S or T; X 4 is D, V, V
  • antibodies of the disclosure can comprise sequences of a heavy chain complementary determining region 1 (HCDR1), an HCDR2, an HCDR3, a light chain complementary determining region 1 (LCDR1), a LCDR2, a LCDR3.
  • HCDR1 heavy chain complementary determining region 1
  • LCDR1 light chain complementary determining region 1
  • LCDR2 a LCDR3.
  • Exemplary CDR amino acid sequences and associated SEQ ID NOs are set forth in Table 1
  • exemplary amino acid sequences for heavy chain variable domains (VH), and light chain variable domains( VL) and associated SEQ ID NOs are set forth in Table 2.
  • any of the antibody or antigen binding fragments thereof of the disclosure comprises: (1) an CDRH1 having a sequence of any one of SEQ ID NOS:45, 46, 47, 48, 49 and 50 or a variant thereof that has a sequence having one amino acid substitution relative to a sequence of any one of SEQ ID NOS: 45, 46, 47, 48, 49 and 50; (2) an CDRH2 having a sequence of any one of SEQ ID NOS: 51, 52, 53, 54, 55 and 56 a variant thereof that has a sequence having one, two, or three amino acid substitutions relative to a sequence of any one of SEQ ID NOS: 51, 52, 53, 54, 55 and 56; (3) an CDRH3 having a sequence of any one of SEQ ID NOS: 57, 58, 59, 60, 61, 62 and 63, or a variant thereof that has a sequence having one or two amino acid substitutions relative to a sequence of any one of SEQ ID NOS: 57, 58,
  • the immunoglobulin heavy chain variable domain comprises a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47,
  • the immunoglobulin heavy chain variable domain comprises: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56; and a CDRH3 comprising the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63.
  • the immunoglobulin light chain variable domain comprises: a CDRL1 comprising the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%
  • the immunoglobulin light chain variable domain comprises: a CDRL1 comprising a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74; and a CDRL3 comprising a sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
  • the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein contains an immunoglobulin heavy chain variable domain with the amino acid sequence set forth in any of SEQ ID NOS:20, 21, 22, 23, 24, 25 or 26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20, 21, 22, 23, 24, 25 or 26.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 20, 21, 22, 23, 24, 25 or 26.
  • the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein contains an immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS:27, 28, 29, 30, 31, 32 or 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27, 28, 29, 30, 31, 32 or 33.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 27, 28, 29, 30, 31, 32 or 33.
  • any of the antibodies or antigen binding fragments thereof further comprises a signal peptide.
  • the immunoglobulin heavy chain variable domain further comprises a signal peptide of any of SEQ ID NOS: 34-38.
  • the immunoglobulin light chain variable domain further comprises a signal peptide of SEQ ID NO: 39-44.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:45 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:45; (2) a CDRH2 having the sequence of SEQ ID NO:51 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:51; (3) a CDRH3 having the sequence of SEQ ID NO:57 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:57.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:64 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:64; (5) a CDRL2 having the sequence of SEQ ID NO:70 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:75 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 75.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45; (2) a CDRH2 having the sequence of SEQ ID NO:51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; (3) a CDRH3 having the sequence of SEQ ID NO:57 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 8
  • the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64; (5) a CDRL2 having the sequence of SEQ ID NO:70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:75 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%,
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:45; (2) an CDRH2 having the sequence of SEQ ID NO: 51; (3) an CDRH3 having the sequence of SEQ ID NO: 57; (4) a CDRL1 having the sequence of SEQ ID NO: 64; (5) a CDRL2 having the sequence of SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:75.
  • the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:20 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:20.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:20.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:27.
  • the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:20 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:20; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:27 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:27.
  • a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:20 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ
  • the antibody or antigen binding fragment thereof can comprise
  • Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:46 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:46; (2) a CDRH2 having the sequence of SEQ ID NO: 52 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:52; (3) a CDRH3 having the sequence of SEQ ID NO:58 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 58.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:65 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 65; (5) a CDRL2 having the sequence of SEQ ID NO:71 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:71; and (6) a CDRL3 having the sequence of SEQ ID NO:76 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:76.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:46 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46; (2) a CDRH2 having the sequence of SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:52; (3) a CDRH3 having the sequence of SEQ ID NO:58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%,
  • the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:65 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 65; (5) a CDRL2 having the sequence of SEQ ID NO:71 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:71; and (6) a CDRL3 having the sequence of SEQ ID NO:76 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%,
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:46; (2) an CDRH2 having the sequence of SEQ ID NO:52; (3) an CDRH3 having the sequence of SEQ ID NO:58; (4) a CDRL1 having the sequence of SEQ ID NO: 65; (5) a CDRL2 having the sequence of SEQ ID NO:71; and (6) a CDRL3 having the sequence of SEQ ID NO:76.
  • the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:21 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:21.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:21.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:28.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:28.
  • the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:21 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:21; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:28 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:28.
  • a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:21 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ
  • the antibody or antigen binding fragment thereof can comprise
  • a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:46, 52 and 58, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 46, 52 and 58 and
  • Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:47 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:47; (2) a CDRH2 having the sequence of SEQ ID NO:53 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:53; (3) a CDRH3 having the sequence of SEQ ID NO:58 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 58.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:66 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 66; (5) a CDRL2 having the sequence of SEQ ID NO: 72 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:72; and (6) a CDRL3 having the sequence of SEQ ID NO:77 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:77.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:47 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47; (2) a CDRH2 having the sequence of SEQ ID NO:53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53; (3) a CDRH3 having the sequence of SEQ ID NO:59 or a sequence of amino acids that exhibits at least 80%, 81%
  • the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:66 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:66; (5) a CDRL2 having the sequence of SEQ ID NO:72 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:72; and (6) a CDRL3 having the sequence of SEQ ID NO:77 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%,
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:47; (2) an CDRH2 having the sequence of SEQ ID NO:53; (3) an CDRH3 having the sequence of SEQ ID NO:59; (4) a CDRL1 having the sequence of SEQ ID NO: 66; (5) a CDRL2 having the sequence of SEQ ID NO:72; and (6) a CDRL3 having the sequence of SEQ ID NO:77.
  • the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:22 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:22.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:22.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:29.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:29.
  • the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:22 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:22; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:29 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:29.
  • a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:22 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ
  • the antibody or antigen binding fragment thereof can comprise (1) a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:47, 53 and 59, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 47, 53 and 59and (2) a light chain variable domain having a CDRL1, a CDRL2, and a CDRL3 of SEQ ID NOS: 66, 72 and 77, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 66, 72 and 77.
  • Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:48 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:48; (2) a CDRH2 having the sequence of SEQ ID NO: 54 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:54; (3) a CDRH3 having the sequence of SEQ ID NO:60 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:60.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:67 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 67; (5) a CDRL2 having the sequence of SEQ ID NO: 73 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:78.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:48 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48; (2) a CDRH2 having the sequence of SEQ ID NO:54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:54; (3) a CDRH3 having the sequence of SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%, 81%
  • the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:67 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:67; (5) a CDRL2 having the sequence of SEQ ID NO:73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%,
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:48; (2) an CDRH2 having the sequence of SEQ ID NO: 54; (3) an CDRH3 having the sequence of SEQ ID NO: 60; (4) a CDRL1 having the sequence of SEQ ID NO: 67; (5) a CDRL2 having the sequence of SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78.
  • the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:23 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:23.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:23.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:30.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:30.
  • the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:20 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:20; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:27 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:27.
  • a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:20 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ
  • the antibody or antigen binding fragment thereof can comprise
  • Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:45 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:45; (2) a CDRH2 having the sequence of SEQ ID NO:51 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:51; (3) a CDRH3 having the sequence of SEQ ID NO:61 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:61.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:64 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:64; (5) a CDRL2 having the sequence of SEQ ID NO:70 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:79 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:79.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45; (2) a CDRH2 having the sequence of SEQ ID NO:51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; (3) a CDRH3 having the sequence of SEQ ID NO:61 or a sequence of amino acids that exhibits at least 80%, 81%
  • the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64; (5) a CDRL2 having the sequence of SEQ ID NO:70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:79 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%,
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:45; (2) an CDRH2 having the sequence of SEQ ID NO:51; (3) an CDRH3 having the sequence of SEQ ID NO:61; (4) a CDRL1 having the sequence of SEQ ID NO: 64; (5) a CDRL2 having the sequence of SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:79.
  • the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:24 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:24.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:24.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:31.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:31.
  • the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:24 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:24; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:31 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 31.
  • the antibody or antigen binding fragment thereof can comprise
  • Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:49 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:49; (2) a CDRH2 having the sequence of SEQ ID NO: 55 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:55; (3) a CDRH3 having the sequence of SEQ ID NO:62 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:62.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:68 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:68; (5) a CDRL2 having the sequence of SEQ ID NO:73 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:78.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:49 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49; (2) a CDRH2 having the sequence of SEQ ID NO:55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:55; (3) a CDRH3 having the sequence of SEQ ID NO:62 or a sequence of amino acids that exhibits at least 80%, 81%, 82%,
  • the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:68 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:68; (5) a CDRL2 having the sequence of SEQ ID NO:73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%,
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:49; (2) an CDRH2 having the sequence of SEQ ID NO:55; (3) an CDRH3 having the sequence of SEQ ID NO:62; (4) a CDRL1 having the sequence of SEQ ID NO:68; (5) a CDRL2 having the sequence of SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78.
  • the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:25 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:25.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:25.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:32, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:32.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:32.
  • the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:35 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:25; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:32 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:32.
  • the antibody or antigen binding fragment thereof can comprise
  • a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:49, 55 and 62, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 49, 55 and 62; and
  • Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:50 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:50; (2) a CDRH2 having the sequence of SEQ ID NO:56 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:56; (3) a CDRH3 having the sequence of SEQ ID NO:63 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:63.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:69 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 69; (5) a CDRL2 having the sequence of SEQ ID NO: 74 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:74; and (6) a CDRL3 having the sequence of SEQ ID NO:80 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:80.
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:50 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:50; (2) a CDRH2 having the sequence of SEQ ID NO:56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56; (3) a CDRH3 having the sequence of SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%, 8
  • the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:69 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:69; (5) a CDRL2 having the sequence of SEQ ID NO:74 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:74; and (6) a CDRL3 having the sequence of SEQ ID NO:80 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%,
  • an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:50; (2) an CDRH2 having the sequence of SEQ ID NO:56; (3) an CDRH3 having the sequence of SEQ ID NO:63; (4) a CDRL1 having the sequence of SEQ ID NO: 69; (5) a CDRL2 having the sequence of SEQ ID NO:74; and (6) a CDRL3 having the sequence of SEQ ID NO:80.
  • the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:26 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:26.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:26.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:33.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:33.
  • the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:26 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:26; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:33 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:33.
  • the antibody or antigen binding fragment thereof can comprise
  • a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:50, 56 and 63, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 50, 56 and 63; and
  • Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
  • an antibody provided herein can comprise a fragment crystallizable region (Fc region), also referred to as an Fc polypeptide herein.
  • An Fc polypeptide is part of each of the two heavy chains in the antibody and can interact with certain cell surface receptors and certain components of the complement system.
  • An Fc polypeptide typically includes the CH2 domain and the CH3 domain, which are immunoglobulin constant region domain polypeptides.
  • the Fc polypeptide in an antibody described herein can be a wild-type Fc polypeptide, e.g., a human IgGl Fc polypeptide.
  • an antibody described herein can comprise a wild-type Fc polypeptide having the sequence of SEQ ID NO:87:
  • an antibody described herein can comprise a variant of the wild-type Fc polypeptide that has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%) identity to the sequence of a wild-type Fc polypeptide (e.g., SEQ ID NO:87) and at least one amino acid substitution relative to the sequence of a wild-type Fc polypeptide (e.g., SEQ ID NO:87).
  • an Fc polypeptide includes one or more modifications (e.g., one or more amino acid substitutions, insertions, or deletions relative to a comparable wildtype Fc region).
  • Antibodies comprising modified Fc polypeptides typically have altered phenotypes relative to antibodies comprising wild-type Fc polypeptides.
  • antibodies comprising modified Fc polypeptides can have altered serum half-life, altered stability, altered susceptibility to cellular enzymes, and/or altered effector function (e.g., as assayed in an NK-dependent or macrophage-dependent assay).
  • an Fc polypeptide in an antibody described herein can include amino acid substitutions that modulate effector function.
  • an Fc polypeptide in an antibody described herein can include amino acid substitutions that reduce or eliminate effector function.
  • Illustrative Fc polypeptide amino acid substitutions that reduce effector function include, but are not limited to, substitutions in a CH2 domain, e.g., at positions 4 and 5 (position numbering relative to the sequence of SEQ ID NO:47) (see, e.g., Lund et al., J Immunol. 147(8):2657-62, 1991).
  • one or both Fc polypeptides in an antibody described herein can comprise L4A and L5A substitutions.
  • Additional Fc polypeptide amino acid substitutions that modulate an effector function include, e.g., substitution at position 99 (position numbering relative to the sequence of SEQ ID NO:5).
  • one or both Fc polypeptides in an antibody described herein can comprise P99G substitution.
  • one or both Fc polypeptides in an antibody described herein can have L4A, L5A, and P99G substitutions.
  • an Fc polypeptide includes one or more modifications that alter (relative to a wild-type Fc polypeptide) the Ratio of Affinities of the modified Fc polypeptide to an activating FcyR (such as FcyRIIA or FcyRIIIA) relative to an inhibiting FcyR (such as FcyRIIB):
  • an antibody herein may have particular use in providing a therapeutic or prophylactic treatment of a disease, disorder, or infection, or the amelioration of a symptom thereof, where an enhanced efficacy of effector cell function (e.g., ADCC) mediated by FcyR is desired, e.g., cancer or infectious disease.
  • ADCC effector cell function
  • a modified Fc region has a Ratio of Affinities less than 1
  • an antibody herein may have particular use in providing a therapeutic or prophylactic treatment of a disease or disorder, or the amelioration of a symptom thereof, where a decreased efficacy of effector cell function mediated by FcyR is desired, e.g., autoimmune or inflammatory disorders.
  • Table 2 lists examples of single, double, triple, quadruple, and quintuple amino acid substitutions in an Fc polypeptide that provide a Ratio of Affinities greater than 1 or less than 1 (see e.g., PCT Publication Nos.
  • antibodies that competitively bind, or are capable of competitively binding may be considered to compete for binding to when the competitor binds to the same general region of as an antibody described herein.
  • an antibody (i.e., competitor antibody) may be considered to compete for binding to when the competitor binds to the exact same region of as an antibody described herein (e.g., exact same peptide (linear epitope) or exact same surface amino acids (conformational epitope)).
  • an antibody i.e., competitor antibody
  • an antibody may be considered capable of competing for binding to when the competitor binds to the same general region of as an antibody described herein (i.e., extracellular region or leucine-rich binding domain) under suitable assay conditions.
  • an antibody i.e., competitor agent
  • an antibody may be considered capable of competing for binding to when the competitor binds to the exact same region of as an antibody described herein (e.g., exact same peptide (linear epitope) or exact same surface amino acids (conformational epitope)) under suitable assay conditions.
  • an antibody i.e., competitor antibody
  • TMPRSS2 an antibody
  • Whether a competitor blocks the binding of one or more antibodies described herein to may be determined using a suitable competition assay or blocking assay, such as, for example, a blocking assay as described in herein.
  • a competitor antibody may block binding of one or more antibodies described herein to in a competition or blocking assay by 50% or more (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, or 100%), and conversely, one or more antibodies described herein may block binding of the competitor antibody to in a competition or blocking assay by about 50% or more (e.g., e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, or 100%).
  • an antibody i.e., competitor antibody
  • an antibody may be considered to compete for binding to TMPRSS2 when the competitor binds to with a similar affinity as one or more antibodies described herein, for example, under suitable assay conditions.
  • an antibody i.e., competitor antibody
  • an affinity that is at least about 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%) of the affinity of one or more antibodies described herein.
  • antibodies that bind to, or are capable of binding to, the same epitope as one or more antibodies described herein are provided herein.
  • antibodies that compete with one or more antibodies described herein for binding to the same epitope e.g., same peptide (linear epitope) or same surface amino acids (conformational epitope)
  • epitope competitors Such antibodies that bind the same epitope may be referred to as epitope competitors.
  • Polyclonal antibodies may be raised in animals (vertebrate or invertebrates, including mammals, birds and fish, including cartilaginous fish) by multiple subcutaneous (sc) or intraperitoneal (ip) injections of a relevant antigen and an adjuvant.
  • animals vertebrate or invertebrates, including mammals, birds and fish, including cartilaginous fish
  • sc subcutaneous
  • ip intraperitoneal
  • Non-protein carriers e.g., colloidal gold
  • a protein or other carrier that is immunogenic in the species to be immunized e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor
  • Non-protein carriers e.g., colloidal gold
  • Animals can be immunized against the antigen, immunogenic conjugates, or derivatives by combining, e.g., 100 pg or 5 pg of the protein or conjugate (for rabbits or mice, respectively) with three volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites.
  • the animals are boosted with one-fifth to one- tenth of the original amount of peptide or conjugate in Freund's complete adjuvant by subcutaneous injection at multiple sites.
  • Seven to 14 days later the animals are bled and the serum is assayed for antibody titer. Animals are boosted until the titer plateaus.
  • the animal is boosted with the conjugate of the same antigen, but conjugated to a different protein and/or through a different cross-linking reagent.
  • Conjugates also can be made in recombinant cell culture as protein fusions. Also, aggregating agents such as alum are suitably used to enhance the immune response.
  • Monoclonal antibodies may be made using a hybridoma, e.g., the hybridoma method first described by Kohler et al., Nature, 256:495 (1975), or may be made by other methods such as recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567).
  • a mouse or other appropriate host animal such as a hamster or macaque monkey, is immunized to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization.
  • lymphocytes may be immunized in vitro.
  • Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (see, e.g., Goding, Monoclonal Antibodies: Principles and Practice, pp.59-103 (Academic Press, 1986)).
  • a suitable fusing agent such as polyethylene glycol
  • the hybridoma cells thus prepared are seeded and grown in a suitable culture medium that may contain one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
  • a suitable culture medium that may contain one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells.
  • the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells.
  • HAT medium hypoxanthine, aminopterin, and thymidine
  • Preferred myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium.
  • preferred myeloma cell lines are murine myeloma lines, such as SP-2 or X63- Ag8-653 cells available from the American Type Culture Collection, Rockville, Md. USA.
  • murine myeloma lines such as SP-2 or X63- Ag8-653 cells available from the American Type Culture Collection, Rockville, Md. USA.
  • Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
  • the binding specificity of monoclonal antibodies produced by hybridoma cells may be determined by immunoprecipitation, by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbant assay (ELISA), or by flow cytometric analysis of cells expressing the membrane antigen.
  • the binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson et al., Anal. Biochem., 107:220 (1980).
  • the clones may be subcloned by limiting dilution procedures and grown by standard methods (see, e.g., Goding, Monoclonal Antibodies: Principles and Practice, pp.59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium.
  • the hybridoma cells may be grown in vivo as ascites tumors in an animal.
  • the monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
  • DNA encoding the monoclonal antibodies can 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 monoclonal antibodies).
  • cDNA may be prepared from mRNA and the cDNA then subjected to DNA sequencing.
  • the hybridoma cells serve as a preferred source of such genomic DNA or RNA for cDNA preparation.
  • the DNA may be placed into expression vectors, which are well known in the art, and which are then transfected into host cells such as E. coll cells, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells.
  • the antibody is a humanized antibody, i.e., an antibody that retains the reactivity of a non-human antibody while being less immunogenic in humans. This can be achieved, for instance, by retaining the non-human CDR regions and replacing the remaining parts of the antibody with their human counterparts.
  • a humanized antibody i.e., an antibody that retains the reactivity of a non-human antibody while being less immunogenic in humans. This can be achieved, for instance, by retaining the non-human CDR regions and replacing the remaining parts of the antibody with their human counterparts. See, e.g., Morrison et al., PNAS USA, 81 :6851-6855 (1984); Morrison and Oi, Adv. Immunol., 44:65-92 (1988); Verhoeyen et al., Science, 239: 1534-1536 (1988); Padlan, Molec. Immun., 28:489-498 (1991); Padlan, Molec.
  • polynucleotides comprising a first sequence coding for humanized immunoglobulin framework regions and a second sequence set coding for the desired immunoglobulin complementarity determining regions can be produced synthetically or by combining appropriate cDNA and genomic DNA segments.
  • Human constant region DNA sequences can be isolated in accordance with well-known procedures from a variety of human cells.
  • the CDRs for producing the immunoglobulins of the present disclosure can be similarly derived from monoclonal antibodies capable of specifically binding to .
  • Amino acid sequence variants of the antibody can be prepared by introducing appropriate nucleotide changes into the antibody DNA, or by peptide synthesis.
  • Such variants include, for example, deletions from, and/or insertions into and/or substitutions of, residues within the amino acid sequences of the antibodies for the examples herein. Any combination of deletion, insertion, and substitution is made to arrive at the final construct, provided that the final construct possesses the desired characteristics.
  • the amino acid changes also may alter post-translational processes of the humanized or variant antibody, such as changing the number or position of glycosylation sites.
  • alanine scanning mutagenesis One method for identification of certain residues or regions of the antibody that are preferred locations for mutagenesis is called “alanine scanning mutagenesis,” as described by, e.g., Cunningham and Wells, Science, 244: 1081-1085 (1989).
  • a residue or group of target residues are identified (e.g., charged residues such as Arg, Asp, His, Lys, and Glu) and replaced by a neutral or negatively charged amino acid (most preferably Ala or poly-Ala) to affect the interaction of the amino acids with antigen.
  • Those amino acid locations demonstrating functional sensitivity to the substitutions then are refined by introducing further or other variants at, or for, the sites of substitution.
  • Amino acid sequence insertions include amino- and/or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues.
  • terminal insertions include an N-terminal methionyl residue or the antibody fused to an epitope tag.
  • Other insertional variants include the fusion of an enzyme or a polypeptide that increases the serum half-life of the antibody to the N- or C- terminus of the antibody.
  • variants Another type of variant is an amino acid substitution variant. These variants have at least one amino acid residue removed from the antibody molecule and a different residue inserted in its place.
  • the sites of greatest interest for substitutional mutagenesis include the hypervariable regions, but FR alterations are also contemplated. Conservative substitutions are preferred, but more substantial changes may be introduced and the products may be screened. Examples of substitutions are listed below:
  • Vai (V) He; Leu; Met; Phe; Ala; Norleucine
  • Substantial modifications in the biological properties of an antibody are accomplished by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain.
  • Naturally occurring residues are divided into groups based on common sidechain properties:
  • Non-conservative substitutions will entail exchanging a member of one of the above classes for another class.
  • Any cysteine residue not involved in maintaining the proper conformation of the antibody also may be substituted, to improve the oxidative stability of the molecule and prevent aberrant crosslinking.
  • cysteine bond(s) may be added to the antibody to improve its stability (particularly where the antibody is an antigen binding fragment such as an Fv fragment).
  • substitutional variant involves substituting one or more hypervariable region residues of a parent antibody.
  • the resulting variant(s) selected for further development will have improved biological properties relative to the parent antibody from which they are generated.
  • a convenient way for generating such substitutional variants is affinity maturation using phage display. Briefly, several hypervariable region sites (e.g., 6-7 sites) are mutated to generate all possible amino acid substitutions at each site.
  • the antibody variants thus generated can be displayed in the monovalent fashion from filamentous phage particles as fusions to the gene III product of M13 packaged within each particle. The phage- displayed variants are then screened for their biological activity (e.g., binding affinity) as herein disclosed.
  • alanine- scanning mutagenesis can be performed to identify hypervariable region residues contributing significantly to antigen binding.
  • Another type of amino acid variant of the antibody alters the original glycosylation pattern of the antibody. By altering is meant deleting one of more carbohydrate moieties found in the antibody, and/or adding one or more glycosylation sites that are not present in the antibody.
  • Glycosylation of antibodies is typically either N-linked and/or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue.
  • the tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the most common recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain.
  • glycosylation sites refers to the attachment of one of the sugars N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5- hydroxyproline or 5-hydroxylysine may also be used.
  • Addition of glycosylation sites to the antibody can be accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). The alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).
  • an antibody is contemplated.
  • technology herein also pertains to immunoconjugates comprising an antibody described herein conjugated to a cytotoxic agent such as a toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant or animal origin, or fragments thereof), or a radioactive isotope (for example, a radioconjugate), or a cytotoxic drug.
  • a cytotoxic agent such as a toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant or animal origin, or fragments thereof), or a radioactive isotope (for example, a radioconjugate), or a cytotoxic drug.
  • Conjugates can be made using a variety of bifunctional protein coupling agents such asN-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis-(p-azidobenzoyl)hexanediamine), bis-diazonium derivatives (such as bis-(p- diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as tolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as l,5-difluoro-2,4-dinitrobenzene).
  • SPDP N-succinimidyl-3-(2-pyridyl
  • any of the antibodies or antigen binding fragments thereof disclosed herein may be formulated as immunoliposomes.
  • Liposomes containing an antibody are prepared by methods know in the art, such as described in Epstein et al., Proc. Natl. Acad. Sci. USA 82:3688 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA 77:4030 (1980); and U.S. Pat. Nos. 4,485,045 and 4,544,545. Liposomes with enhanced circulation time are disclosed in U.S. Pat. No. 5,013,556.
  • liposomes can be generated by the reverse phase evaporation method with a lipid composition comprising phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to yield liposomes with the desired diameter.
  • Fab' fragments of an antibody provided herein can be conjugated to the liposomes as described in Martin et al., J. Biol. Chem. 257:286-288 (1982) via a disulfide interchange reaction. Another active ingredient is optionally contained within the liposome.
  • Enzymes or other polypeptides can be covalently bound to an antibody by techniques well known in the art such as the use of the heterobifunctional cross-linking reagents discussed above.
  • fusion proteins comprising at least the antigen binding region of an antibody provided herein linked to at least a functionally active portion of an enzyme can be constructed using recombinant DNA techniques well known in the art (see, e.g., Neuberger et al., Nature 312:604-608 (1984)).
  • an antigen binding fragment rather than an intact antibody, to increase penetration of target tissues and cells, for example.
  • any of the antibodies or antigen fragments thereof disclosed herein are conjugated or hybridized to an oligonucleotide.
  • the oligonucleotide includes a sample barcode sequence, a binding site for a primer and an anchor.
  • the oligonucleotide can be conjugated or hybridized to any of the detectable markers or labels disclosed herein.
  • the oligonucleotide is a polymeric sequence.
  • oligonucleotide and “polynucleotide” are used interchangeably to refer to a single-stranded multimer of nucleotides from about 2 to about 500 nucleotides in length.
  • any of the oligonucleotides described herein can be synthetic, made enzymatically (e.g., via polymerization), or using a “split-pool” method.
  • any of the oligonucleotides described herein can include ribonucleotide monomers (i.e., can be oligoribonucleotides) and/or deoxyribonucleotide monomers (i.e., oligodeoxyribonucleotides).
  • any of the oligonucleotides described herein can include a combination of both deoxyribonucleotide monomers and ribonucleotide monomers in the oligonucleotide (e.g., random or ordered combination of deoxyribonucleotide monomers and ribonucleotide monomers).
  • the oligonucleotide can be 4 to 10, 10 to 20, 21 to 30, 31 to 40, 41 to 50, 51 to 60, 61 to 70, 71 to 80, 80 to 100, 100 to 150, 150 to 200, 200 to 250, 250 to 300, 300 to 350, 350 to 400, or 400-500 nucleotides in length.
  • any of the oligonucleotides described herein can include one or more functional moieties that are attached (e.g., covalently or non-covalently) to another structure.
  • any of the oligonucleotides described herein can include one or more detectable labels (e.g., a radioisotope or fluorophore).
  • the anchor is a defined polymer, e.g., a polynucleotide or oligonucleotide sequence, which is designed to hybridize to a complementary oligonucleotide sequence.
  • the anchor is designed for the purpose of generating a double stranded construct oligonucleotide sequence.
  • the anchor is positioned at the 3’ end of the construct oligonucleotide sequence. In other embodiments, the anchor is positioned at the 5’ end of the construct oligonucleotide sequence.
  • Each anchor is specific for its intended complementary sequence.
  • the sample barcode sequence is a polymer, e.g., a polynucleotide, which when it is a functional element, is specific for a single ligand.
  • the sample barcode sequence can be used for identifying a particular cell or substrate, e.g., Drop-seq microbead.
  • the sample barcode sequence can be formed of a defined sequence of DNA, RNA, modified bases or combinations of these bases, as well as any other polymer defined above.
  • the sample barcode sequence is about 2 to 4 monomeric components, e.g., nucleotide bases, in length.
  • the barcode is at least about 1 to 100 monomeric components, e.g., nucleotides, in length.
  • the barcode is formed of a sequence of at least 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,
  • the sample barcode sequence is a particular barcode that can be unique relative to other barcodes.
  • sample barcode sequences can have a variety of different formats.
  • sample barcode sequences can include polynucleotide barcodes, random nucleic acid and/or amino acid sequences, and synthetic nucleic acid and/or amino acid sequences.
  • a sample barcode sequence can be attached to an analyte or to another moiety or structure in a reversible or irreversible manner.
  • a sample barcode sequences can be added to, for example, a fragment of a deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) sample before or during sequencing of the sample.
  • Sample barcode sequences can allow for identification and/or quantification of individual sequencing-reads (e.g., a barcode can be or can include a unique molecular identifier or “UMI”).
  • Sample barcode sequences can spatially-resolve molecular components found in biological samples, for example, at single-cell resolution (e.g., a barcode can be or can include a “spatial barcode”).
  • a barcode includes both a UMI and a spatial barcode.
  • a barcode includes two or more sub-barcodes that together function as a single barcode.
  • a polynucleotide barcode can include two or more polynucleotide sequences (e.g., sub-barcodes) that are separated by one or more non-barcode sequences.
  • the binding site for a primer is a functional component of the oligonucleotide which itself is an oligonucleotide or polynucleotide sequence that provides an annealing site for amplification of the oligonucleotide.
  • the binding site for a primer can be formed of polymers of DNA, RNA, PNA, modified bases or combinations of these bases, or polyamides, etc.
  • the binding site for a primer is about 10 of such monomeric components, e.g., nucleotide bases, in length.
  • the binding site for a primer is at least about 5 to 100 monomeric components, e.g., nucleotides, in length.
  • the binding site for a primer is formed of a sequence of at least 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, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 80, 91, 92, 93, 94, 95, 96, 97, 98, 99 or up to 100 monomeric components, e.g., nucleic acids.
  • the binding site for a primer can be a generic sequence suitable as a annealing site for a variety of amplification technologies.
  • Amplification technologies include, but are not limited to, DNA-polymerase based amplification systems, such as polymerase chain reaction (PCR), real-time PCR, loop mediated isothermal amplification (LAMP, MALBAC), strand displacement amplification (SDA), multiple displacement amplification (MDA), recombinase polymerase amplification (RPA) and polymerization by any number of DNA polymerases (for example, T4 DNA polymerase, Sulfulobus DNA polymerase, Klenow DNA polymerase, Bst polymerase, Phi29 polymerase) and RNA-polymerase based amplification systems (such as T7-, T3-, and SP6-RNA- polymerase amplification), nucleic acid sequence based amplification (NASBA), selfsustained sequence replication (3 SR), rolling circle amplification (RCA),
  • Methods for conjugating or hybridizing an oligonucleotide can be performed in a manner set forth in WO/2018/144813, WO/2017/018960, WO/2018/089438, WO/2014/182528, WO/2018/026873, WO/2021/188838.
  • a modification can optionally be introduced into the antibodies (e.g., within the polypeptide chain or at either the N- or C-terminal), e.g., to extend in vivo half-life, such as PEGylation or incorporation of long-chain polyethylene glycol polymers (PEG).
  • PEG polyethylene glycol polymers
  • Introduction of PEG or long chain polymers of PEG increases the effective molecular weight of the polypeptides, for example, to prevent rapid filtration into the urine.
  • a lysine residue in the sequence is conjugated to PEG directly or through a linker.
  • Such linker can be, for example, a Glu residue or an acyl residue containing a thiol functional group for linkage to the appropriately modified PEG chain.
  • An alternative method for introducing a PEG chain is to first introduce a Cys residue at the C-terminus or at solvent exposed residues such as replacements for Arg or Lys residues. This Cys residue is then site- specifically attached to a PEG chain containing, for example, a maleimide function.
  • Methods for incorporating PEG or long chain polymers of PEG are known in the art (described, for example, in Veronese, F. M., et al., Drug Disc. Today 10: 1451-8 (2005); Greenwald, R. B., et al., Adv. Drug Deliv. Rev. 55: 217-50 (2003); Roberts, M. J., et al., Adv. Drug Deliv. Rev., 54: 459-76 (2002)), the contents of which are incorporated herein by reference.
  • Covalent modifications of an antibody are also included within the scope of this technology. For example, modifications may be made by chemical synthesis or by enzymatic or chemical cleavage of an antibody. Other types of covalent modifications of an antibody are introduced into the molecule by reacting targeted amino acid residues of the antibody with an organic derivatizing agent that is capable of reacting with selected side chains or the N- or C- terminal residues.
  • Example covalent modifications of polypeptides are described in U.S. Pat. No. 5,534,615, specifically incorporated herein by reference.
  • a preferred type of covalent modification of the antibody comprises linking the antibody to one of a variety of non- proteinaceous polymers, e.g., polyethylene glycol, polypropylene glycol, or polyoxyalkylenes, in the manner set forth in, e.g., U.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337. VII. Nucleic acids, vectors, host cells, and recombinant methods
  • the disclosure also provides isolated nucleic acids encoding an antibody, vectors and host cells comprising the nucleic acid, and recombinant techniques for the production of the antibody.
  • a nucleic acid herein may include one or more subsequences, each referred to as a polynucleotide.
  • nucleic acids e.g., isolated nucleic acids
  • a nucleic acid encodes an immunoglobulin heavy chain variable domain of an antibody provided herein.
  • a nucleic acid encodes an immunoglobulin light chain variable domain of an antibody provided herein.
  • a nucleic acid encodes an immunoglobulin heavy chain variable domain and an immunoglobulin light chain variable domain of an antibody provided herein.
  • a nucleic acid comprises a nucleotide sequence that encodes an amino acid sequence of any one of SEQ ID NOS: 6-12 or 13-19.
  • the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in any of SEQ ID NOs: 6-12. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 6. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 7. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 8. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 9.
  • the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 10. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 11. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 12.
  • the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in any of SEQ ID NOs: 13-19. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 13. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 14. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 15. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 16.
  • the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 17. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 18. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 19.
  • nucleotide sequence that encodes the immunoglobulin heavy chain variable domain and the immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any of any of the antibodies or antigen binding fragments provided herein.
  • the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 6; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 13.
  • the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 7; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 14.
  • the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 11; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 18. In some of any embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 12; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 19.
  • the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in any of SEQ ID NOs:6-12; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in any of SEQ ID NOs: 13-19.
  • a nucleic acid encoding the antibody may be isolated and inserted into a replicable vector for further cloning (amplification of the DNA) or for expression.
  • an antibody may be produced by homologous recombination.
  • DNA encoding an antibody can 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).
  • Many vectors are available.
  • the vector components generally include, but are not limited to, one or more of the following: a signal sequence, and origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence.
  • Suitable host cells for cloning or expressing DNA in vectors herein can be prokaryote, yeast, or higher eukaryote cells.
  • Suitable prokaryotes for this purpose include eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as Escherichia, e.g., E. coli, Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium, Serratia, e.g., Serratia marcescans, and Shigella, as well as Bacilli such as //. subtilis and B.
  • Enterobacteriaceae such as Escherichia, e.g., E. coli, Enterobacter, Erwinia, Klebsiella, Proteus
  • Salmonella e.g., Salmonella typhimurium
  • Serratia e.g., Serratia marc
  • E. coli 294 ATCC 31,446
  • E. coli B E. coli X1776
  • E. coli W3110 ATCC 27,325
  • eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for antibody-encoding vectors.
  • Saccharomyces cerevisiae, or common baker’s yeast is the most commonly used among lower eukaryotic host microorganisms.
  • a number of other genera, species, and strains are commonly available and useful herein, such as Schizosaccharomyces pom be , Kluyveromyces hosts such as, e.g., K. lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K.
  • waltii ATCC 56,500
  • K. drosophilarum ATCC 36,906
  • K. thermotolerans K. marxianus: yarrowia (EP 402,226); Pichia pastoris (EP 183,070); Candida,' Trichoderma reesia (EP 244,234)
  • Neurospora crassa Schwanniomyces such as Schwanniomyces occidenlahs and filamentous fungi such as, e.g., Neurospora, Penicillium, Tolypocladium, and Aspergillus hosts such as A. nidulans and A. niger.
  • Suitable host cells for the expression of antibodies can also be derived from multicellular organisms.
  • invertebrate cells include plant and insect cells.
  • Numerous baculoviral strains and variants and corresponding permissive insect host cells from hosts such as Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruitfly), and Bombyx mori (silk moth) have been identified.
  • a variety of viral strains for transfection are publicly available, e.g., the L-l variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used as the virus herein according to the present technology, particularly for transfection of Spodoptera frugiperda cells.
  • Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco can also be utilized as hosts.
  • Suitable host cells for the expression of antibodies also may include vertebrate cells (e.g., mammalian cells). Vertebrate cells may be propagated in culture (tissue culture). Examples of useful mammalian host cell lines include monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BEK, ATCC CCL 10); Chinese hamster ovary cells/-DHFR (CHO, Urlaub et al., Proc. Natl. Acad. Sci.
  • mice Sertoli cells TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3 A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci.
  • Host cells may be transformed with the above-described expression or cloning vectors for antibody production and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.
  • Host cells used to produce antibodies provided herein may be cultured in a variety of media. Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPMI-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing the host cells.
  • any of the media described in Ham et al., Meth. Enz. 58:44 (1979), Barnes et al., Anal. Biochem.102:255 (1980), U.S. Pat. Nos. 4,767,704; 4,657,866; 4,927,762; 4,560,655; or 5,122,469; WO 90/03430; WO 87/00195; or U.S. Pat. Re. 30,985 may be used as culture media for the host cells.
  • any of these media may be supplemented as necessary with hormones and/or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as GENTAMYCINTM), trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art.
  • the culture conditions such as temperature, pH, and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.
  • antibodies can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the antibody is produced intracellularly, as a first step, the particulate debris, either host cells or lysed fragments, is removed, for example, by centrifugation or ultrafiltration. Carter et al., Bio/Technology 10: 163-167 (1992) describe a procedure for isolating antibodies that are secreted to the periplasmic space of E. coli. Briefly, cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min.
  • sodium acetate pH 3.5
  • EDTA EDTA
  • PMSF phenylmethylsulfonylfluoride
  • Cell debris can be removed by centrifugation.
  • supernatants from such expression systems are generally first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit.
  • a protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants.
  • the antibody composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being the preferred purification technique.
  • protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc domain that is present in the antibody.
  • Protein A can be used to purify antibodies that are based on human heavy chains (Lindmark et al., J. Immunol. Meth. 62: 1-13 (1983)).
  • Protein G is recommended for all mouse isotypes and for human y3 (Guss et al., EMBO J. 5: 15671575 (1986)).
  • the matrix to which the affinity ligand is attached is most often agarose, but other matrices are available.
  • Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose.
  • the antibody comprises a CH3 domain
  • Bakerbond ABX.TM. resin J. T. Baker, Phillipsburg, N.J. is useful for purification.
  • the mixture comprising the antibody of interest and contaminants may be subjected to low pH hydrophobic interaction chromatography using an elution buffer at a pH between, e.g., about 2.5-4.5, and may be performed at low salt concentrations (e.g., from about 0-0.2 5M salt).
  • the present disclosure provides antibodies and related compositions, which may be useful for elimination of -expressing pathogens from the body, for example, and for identification and quantification of the number of TMPRSS2-expressing pathogens in biological samples, for example.
  • any of the antibodies or antigen binding fragments thereof may be formulated in a pharmaceutical composition that is useful for a variety of purposes, including the treatment of diseases or disorders.
  • Pharmaceutical compositions comprising one or more antibodies may be administered using a pharmaceutical device to a patient in need thereof, and according to one embodiment of the technology, kits are provided that include such devices. Such devices and kits may be designed for routine administration, including selfadministration, of the pharmaceutical compositions herein.
  • Therapeutic formulations of an antibody may be prepared for storage by mixing the agent or antibody having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers (Remington’s Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions.
  • Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues ) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine,
  • the disease or disorder is associated with TMPRSS2 expression. In some embodiments, the disease or disorder is associated with aberrant TMPRSS2 expression. In some embodiments, the disease or disorder is associated with Natural Killer (NK), alpha beta T cells, gamma delta T cells, CD8+ T cells, monocytes, or dendritic cells. In some embodiments, the disease or disorder is associated with Natural Killer (NK) cells. In some embodiments, the disease or disorder is associated with alpha beta T cells. In some embodiments, the disease or disorder is associated with gamma delta T cells. In some embodiments, the disease or disorder is associated with CD8+ T cells. In some embodiments, the disease or disorder is associated with monocytes. In some embodiments, the disease or disorder is associated with dendritic cells.
  • NK Natural Killer
  • alpha beta T cells gamma delta T cells
  • CD8+ T cells gamma delta T cells.
  • the disease or disorder is associated with CD8+ T cells.
  • the disease or disorder is associated with
  • the disease or disorder is a cancer, an infectious disease, or an autoimmune disorder.
  • the disease or disorder is a cancer.
  • the cancer is metastatic melanoma, a solid tumor, bladder cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma, hepatic metastasis of colonic origin, papillary thyroid carcinoma, acute myeloid leukemia, or asymptomatic myeloma.
  • the disease or disorder is an infectious disease.
  • the infectious disease is human immunodeficiency virus (HIV), chronic hepatitis C, cytomegalovirus, or hantavirus.
  • HIV human immunodeficiency virus
  • chronic hepatitis C chronic hepatitis C
  • cytomegalovirus cytomegalovirus
  • hantavirus hantavirus
  • the disease or disorder is an autoimmune disorder.
  • the autoimmune disorder is Chrohn’s disease, multiple sclerosis, systemic sclerosis, ocular myasthenia gravis, psoriasis or rheumatoid arthritis.
  • any of the antibodies or antigen binding fragments thereof described herein can be used to decrease the production of androgenic hormones in prostate cancer cells.
  • any of the antibodies or antigen binding fragments thereof described herein can be used to inhibit proteolytic cleavage of ACE2 receptor.
  • any of the antibodies or antigen binding fragments thereof described herein can be used to inhibit or reduce cleavage of coronavirus spike glycoproteins. In some embodiments, any of the antibodies or antigen binding fragments thereof described herein can be used to inhibit or reduce viral uptake into a host cell.
  • Formulations herein may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Such molecules are suitably present in combination in amounts that are effective for the purpose intended.
  • Formulations for in vivo administration generally are sterile. This may be accomplished for instance by filtration through sterile filtration membranes, for example.
  • sustained-release preparations may be prepared. Suitable examples of sustained- release preparations include semipermeable matrices of solid hydrophobic polymers containing the agent/antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly (vinyl alcohol)), polylactides (U.S. Pat. No.
  • copolymers of L-glutamic acid and gamma ethyl-L-glutamate non-degradable ethylene-vinyl acetate
  • degradable lactic acid-glycolic acid copolymers such as the Lupron Depot® (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate)
  • poly-D-(-)-3 -hydroxybutyric acid While polymers such as such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods.
  • encapsulated agents/antibodies When encapsulated agents/antibodies remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37 °C, resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S-S bond formation through thiol-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.
  • antibodies provided herein are administered to a mammal, e.g., a human, in a pharmaceutically acceptable dosage form such as those discussed above, including those that may be administered to a human intravenously as a bolus or by continuous infusion over a period of time, or by intramuscular, intraperitoneal, intra- cerebrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral, topical, or inhalation routes.
  • the appropriate dosage of agent or antibody will depend on the type of disease to be treated, as defined above, the severity and course of the disease, whether the antibody is administered for preventative or therapeutic purposes, previous therapy, the patient’s clinical history and response to the antibody, and the discretion of the attending physician.
  • the antibody is suitably administered to the patient at one time or over a series of treatments.
  • about 1 pg/kg to about 50 mg/kg (e.g., 0.1-20 mg/kg) of antibody may be an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion.
  • a typical daily or weekly dosage might range from about 1 pg/kg to about 20 mg/kg or more, depending on the factors mentioned above.
  • the treatment is repeated until a desired suppression of disease symptoms occurs.
  • other dosage regimens may be useful.
  • the progress of this therapy is easily monitored by conventional techniques and assays, including, for example, radiographic imaging. Detection methods using the antibody to determine TMPRSS2 levels in bodily fluids or tissues may be used in order to optimize patient exposure to the therapeutic antibody.
  • a composition comprising an antibody herein can be administered as a monotherapy, and in some embodiments, the composition comprising the antibody can be administered as part of a combination therapy.
  • the effectiveness of the antibody in preventing or treating diseases may be improved by administering the antibody serially or in combination with another drug that is effective for those purposes, such as a chemotherapeutic drug for treatment of cancer or a microbial infection.
  • the antibody may serve to enhance or sensitize cells to chemotherapeutic treatment, thus permitting efficacy at lower doses and with lower toxicity.
  • Certain combination therapies include, in addition to administration of the composition comprising an antibody that reduces the number of -expressing cells, delivering a second therapeutic regimen selected from the group consisting of a chemotherapeutic agent, radiation therapy, surgery, and a combination of any of the foregoing.
  • a second therapeutic regimen selected from the group consisting of a chemotherapeutic agent, radiation therapy, surgery, and a combination of any of the foregoing.
  • Such other agents may be present in the composition being administered or may be administered separately.
  • the antibody may be suitably administered serially or in combination with the other agent or modality, e.g., chemotherapeutic drug or radiation for treatment of cancer, infection, and the like, or an immunosuppressive drug.
  • diagnostic reagents comprising an antibody described herein.
  • antibodies provided herein may be used to detect and/or purify TMPRSS2 from bodily fluid(s) or tissues.
  • methods for detecting TMPRSS2. For example, a method may comprise contacting a sample (e.g., a biological sample known or suspected to contain ) with an antibody provided herein, and, if the sample contains TMPRSS2, detecting TMPRSS2: antibody complexes.
  • reagents comprising an antibody described herein and methods for detecting for research purposes.
  • an antibody comprises a detectable marker or label.
  • an antibody is conjugated to a detectable marker or label.
  • an antibody may be labeled with a detectable moiety. Numerous labels are available which are generally grouped into the following categories:
  • Radioisotopes such as 35S, 14C, 1251, 3H, and 1311.
  • the antibody can be labeled with the radioisotope using the techniques described in Current Protocols in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-Interscience, New York, N.Y., Pubs. (1991), for example, and radioactivity can be measured using scintillation counting.
  • Fluorescent labels such as rare earth chelates (europium chelates) or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin, Texas Red and Brilliant VioletTM are available.
  • the fluorescent labels can be conjugated to the antibody using the techniques disclosed in Current Protocols in Immunology, supra, for example. Fluorescence can be quantified using a flow cytometer, imaging microscope or fluorimeter.
  • the enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above.
  • the chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light that can be measured (using a chemilluminometer, for example) or donates energy to a fluorescent acceptor.
  • enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase), luciferin, 2,3- dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, beta-galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclicoxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like.
  • luciferases e.g., firefly luciferase and bacterial luciferase
  • luciferin 2,3- dihydrophthalazin
  • HRP Horseradish peroxidase
  • OPD orthophenylene diamine
  • TMB 3, 3', 5,5'- tetramethyl benzidine hydrochloride
  • P-D -galactosidase P-D-Gal
  • a chromogenic substrate e.g., p-nitrophenyl-P-D- galactosidase
  • fluorogenic substrate 4-methylumbelliferyl-P-D-galactosidase
  • the label is indirectly conjugated with the agent or antibody.
  • an antibody can be conjugated with biotin and any of the three broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the antibody in this indirect manner.
  • the antibody is conjugated with a small hapten (e.g., digoxin) and one of the different types of labels mentioned above is conjugated with an anti-hapten antibody (e.g., anti-digoxin antibody).
  • a small hapten e.g., digoxin
  • an anti-hapten antibody e.g., anti-digoxin antibody
  • antibody or antigen binding fragments thereof need not be labeled, and the presence thereof can be detected, e.g., using a labeled antibody which binds to an antibody.
  • an antibody herein is immobilized on a solid support or substrate.
  • an antibody herein is non-diffusively immobilized on a solid support (e.g., the antibody does not detach from the solid support).
  • a solid support or substrate can be any physically separable solid to which an antibody can be directly or indirectly attached including, but not limited to, surfaces provided by microarrays and wells, and particles such as beads (e.g., paramagnetic beads, magnetic beads, microbeads, nanobeads), microparticles, and nanoparticles.
  • Solid supports also can include, for example, chips, columns, optical fibers, wipes, filters (e.g., flat surface filters), one or more capillaries, glass and modified or functionalized glass (e.g., controlled-pore glass (CPG)), quartz, mica, diazotized membranes (paper or nylon), polyformaldehyde, cellulose, cellulose acetate, paper, ceramics, metals, metalloids, semiconductive materials, quantum dots, coated beads or particles, other chromatographic materials, magnetic particles; plastics (including acrylics, polystyrene, copolymers of styrene or other materials, polybutylene, polyurethanes, TEFLONTM, polyethylene, polypropylene, polyamide, polyester, polyvinylidenedifluoride (PVDF), and the like), polysaccharides, nylon or nitrocellulose, resins, silica or silica-based materials including silicon, silica gel, and modified silicon, Sephadex®, Sepharose®, carbon, metals (
  • the solid support or substrate may be coated using passive or chemically-derivatized coatings with any number of materials, including polymers, such as dextrans, acrylamides, gelatins or agarose. Beads and/or particles may be free or in connection with one another (e.g., sintered).
  • a solid support or substrate can be a collection of particles.
  • the particles can comprise silica, and the silica may comprise silica dioxide.
  • the silica can be porous, and in certain embodiments the silica can be non-porous.
  • the particles further comprise an agent that confers a paramagnetic property to the particles.
  • the agent comprises a metal
  • the agent is a metal oxide, (e.g., iron or iron oxides, where the iron oxide contains a mixture of Fe2+ and Fe3+).
  • An antibody may be linked to a solid support by covalent bonds or by non-covalent interactions and may be linked to a solid support directly or indirectly (e.g., via an intermediary agent such as a spacer molecule or biotin).
  • Antibodies and antigen binding fragments thereof provided herein may be employed in any known assay method, such as flow cytometry, immunohistochemistry, immunofluorescence, mass cytometry (e.g., Cytof instrument), competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158 (CRC Press, Inc. 1987). Flow cytometry and mass cytometry assays generally involve the use of a single primary antibody to specifically identify the presence of the target molecule expressed on the surface of a dispersed suspension of individual cells.
  • the dispersed cells are typically obtained from a biological fluid sample, e.g., blood, but may also be obtained from a dispersion of single cells prepared from a solid tissue sample such as spleen or tumor biopsy.
  • the primary antibody may be directly conjugated with a detectable moiety, e.g., a fluorophore such as phycoerythrin for flow cytometry or a heavy metal chelate for mass cytometry.
  • the primary antibody may be unlabeled or labeled with an undetectable tag such as biotin, and the primary antibody is then detected by a detectably labeled secondary antibody that specifically recognizes the primary antibody itself or the tag on the primary antibody.
  • the labeled cells are then analyzed in an instrument capable of single cell detection, e.g., flow cytometer, mass cytometer, fluorescence microscope or brightfield light microscope, to identify those individual cells in the dispersed population or tissue sample that express the target recognized by the primary antibody.
  • an instrument capable of single cell detection e.g., flow cytometer, mass cytometer, fluorescence microscope or brightfield light microscope.
  • flow cytometry principles may be found in, e.g., Shapiro, Practical Flow Cytometry, 4th Edition, Wiley, 2003.
  • Sandwich assays involve the use of two antibodies, each capable of binding to a different immunogenic portion, or epitope, of the protein that is detected.
  • the test sample analyte is bound by a first antibody that is immobilized on a solid support, and thereafter a second antibody binds to the analyte, thus forming an insoluble three-part complex.
  • the second antibody may itself be labeled with a detectable moiety (direct sandwich assays) or may be measured using an anti-immunoglobulin antibody that is labeled with a detectable moiety (indirect sandwich assay).
  • one type of sandwich assay is an ELISA assay, in which case the detectable moiety is an enzyme.
  • the target cell population may be attached to the solid support using antibodies first attached to the support and that recognize different cell surface proteins. These first antibodies capture the cells to the support.
  • TMPRSS2 on the surface of the cells can then be detected by adding any of the anti-TMPRSS2 antibodies or antigen-binding fragments thereof described herein to the captured cells and detecting the amount of the anti-TMPRSS2 antibody or antigen binding fragment thereof attached to the cells.
  • fixed and permeabilized cells may be used, and in such instances, both surface TMPRSS2 and intracellular TMPRSS2 may be detected.
  • any of the antibodies or antigen binding fragments thereof provided herein are formulated for immunohistochemical analysis.
  • immunohistochemical analysis includes the use of samples.
  • immunohistochemical analysis includes the use of blood and/or tissue samples.
  • the sample may be fresh or frozen or may be embedded in paraffin and fixed with a preservative such as formalin.
  • the sample is a formalin-fixed paraffin-embedded (FFPE) sample.
  • FFPE formalin-fixed paraffin-embedded
  • the FFPE sample is saturated with formalin (i.e. formaldehyde) and then embedded in a block of paraffin wax.
  • the FFPE sample is stable at room temperature.
  • all of the structures in the FFPE sample are preserved.
  • the intracellular and surface proteins in the FFPE sample are preserved.
  • the mRNA in the FFPE sample is preserved.
  • the mRNA, intracellular and surface proteins in the FFPE sample are preserved.
  • the surface proteins in the FFPE sample are denatured.
  • any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting TMPRSS2 in a formalin-fixed paraffin-embedded sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting TMPRSS2 on the surface of a in a formalin-fixed paraffin-embedded sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting intracellular TMPRSS2 in a formalin-fixed paraffin-embedded sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting intracellular TMPRSS2, and TMPRSS2 on the surface of a formalin-fixed paraffin-embedded sample.
  • the sample is a fresh sample that has been frozen. In some embodiments, the sample is a fresh sample that has been cryogenically frozen. In some embodiments, the sample is flash frozen. In some embodiments, the sample if flash frozen and stored at 80°C. In some embodiments, all of the structures in the flash frozen sample are preserved. In some embodiments, the intracellular and surface proteins in the flash frozen sample are preserved. In some embodiments, the mRNA in the flash frozen sample is preserved. In some embodiments, the mRNA, intracellular and surface proteins in the flash frozen sample are preserved. In some embodiments, the surface proteins in the flash frozen sample are denatured.
  • any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting TMPRSS2 in a frozen sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting TMPRSS2 on the surface of a frozen sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting intracellular TMPRSS2 in a frozen sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting intracellular TMPRSS2, and TMPRSS2 on the surface of a frozen sample.
  • the antibodies herein also may be used for in vivo diagnostic assays.
  • the antibody is labeled with a radionuclide (such as 11 Un, 99Tc, 14C, 1311, 1251, 3H, 32P, or 35S) so that the bound target molecule can be localized using immunoscintillography.
  • a radionuclide such as 11 Un, 99Tc, 14C, 1311, 1251, 3H, 32P, or 35S
  • antibodies and methods for detecting TMPRSS2 are provided herein.
  • the biological sample is a solid tissue, fluid, or cell.
  • the TMPRSS2 is detected on the surface of the cell.
  • the TMPRSS2 is detected intracellularly.
  • the detection of TMPRSS2 is in vitro.
  • the detection of TMPRSS2 is in vivo.
  • the solid tissue may comprise solid tissue from one or more of adipose tissue, bladder, bone, brain breast cervix, endothelium, gallbladder, kidney, liver, lung, lymph, ovary, prostate, salivary gland, stomach, testis, thyroid, urethra, uterus, vagina, and vulva.
  • the fluid comprises one or more of amniotic fluid, bile, blood, breast milk, breast fluid, cerebrospinal fluid, lavage fluid, lymphatic fluid, mucous, plasma, saliva, semen, serum, spinal fluid, sputum, tears, umbilical cord blood, urine, and vaginal fluid.
  • the sample comprises immune cells.
  • the sample comprises a heterogeneous population of immune cells.
  • the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs).
  • pDCs plasmacytoid dendritic cells
  • mDCs myeloid dendritic cells
  • mast cells eosinophils, basophils, natural killer cells
  • PBMCs peripheral blood mononuclear cells
  • any of the antibodies or antigen binding fragments thereof provided herein can be used in the characterization of single cells by measurement of gene-expression levels and cellular proteins.
  • known single cell sequencing platforms suitable for integration with the antibodies or antigen binding fragments thereof described herein is the Drop-seq method, including, but not limited to, microfluidic, plate- based, or microwell, Seq-WellTM method and adaptations of the basic protocol, and InDropTM method.
  • a single cell sequencing platform suitable for integration with the antibodies or antigen binding fragments thereof described herein is lOx genomics single cell 3' solution or single cell V(D)J solution, either run on Chromium controller, or dedicated Chromium single cell controller.
  • sequencing methods include Wafergen iCell8TM method, Microwell-seq method, Fluidigm CITM method and equivalent single cell products.
  • Still other known sequencing protocols useful with the antibodies or antigen binding fragments thereof described herein include BD ResolveTM single cell analysis platform and ddSeq (from Illumina® Bio-Rad® SureCellTM WTA 3' Library Prep Kit for the ddSEQTM System, 2017, Pub. No. 1070-2016-014-B, Illumina Inc., Bio-Rad Laboratories, Inc.).
  • the antibodies or antigen binding fragments thereof described herein are useful with combinatorial indexing based approaches (sci-RNA-seqTM method or SPLiT-seqTM method) and Spatial Transcriptomics, or comparable spatially resolved sequencing approaches.
  • combinatorial indexing based approaches sci-RNA-seqTM method or SPLiT-seqTM method
  • Spatial Transcriptomics or comparable spatially resolved sequencing approaches.
  • the methods and compositions described herein can also be used as an added layer of information on standard index sorting (FACS) and mRNA-sequencing- based approaches.
  • any of the antibodies or antigen binding fragments thereof described herein can be used to detect the presence, absence or amount of the various nucleic acids, proteins, targets, oligonucleotides, amplification products and barcodes described herein.
  • the biological sample is from a healthy subject. In some embodiments, the sample is from a subject with a disease or condition. In some embodiments, the detection of TMPRSS2 indicates the presence or absence of a disease or disorder. In some embodiments, the disease or disorder is a cancer, an autoimmune disorder, an inflammatory disorder, a neurologic disorder, or an infection.
  • the cancer is the cancer is acute myeloid leukemia, acute lymphoblastic leukemia, colorectal, ovarian, gynecologic, liver, glioblastoma, Hodgkin lymphoma, chronic lymphocytic leukemia, esophagus, gastric, pancreas, colon, kidney, head and neck, lung and melanoma.
  • the disease or disorder is associated with TMPRSS2 expression, In some embodiments, the disease or disorder is associated with aberrant TMPRSS2 expression. In some embodiments, the disease or disorder is associated with Natural Killer (NK), alpha beta T cells, gamma delta T cells, CD8+ T cells, monocytes, or dendritic cells. In some embodiments, the disease or disorder is associated with Natural Killer (NK) cells. In some embodiments, the disease or disorder is associated with alpha beta T cells. In some embodiments, the disease or disorder is associated with gamma delta T cells. In some embodiments, the disease or disorder is associated with CD8+ T cells. In some embodiments, the disease or disorder is associated with monocytes.
  • NK Natural Killer
  • alpha beta T cells In some embodiments, the disease or disorder is associated with gamma delta T cells.
  • the disease or disorder is associated with CD8+ T cells. In some embodiments, the disease or disorder is associated with monocytes.
  • the disease or disorder is associated with dendritic cells.
  • the disease or disorder is chosen from non-viral cancers, virus-associated cancers, cancers associated with HBV infection, cancers associated with Epstein-Barr virus (EBV) infection, cancers associated with polyomavirus infection, erythema nodosum leprosum (ENL), autoimmune diseases, autoimmune inflammation, autoimmune thyroid diseases, B-cell lymphoma, T-cell lymphoma, acute myeloid leukemia, Hodgkin's Disease, acute myelogenous leukemia, acute myelomonocytic leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, B cell large cell lymphoma, malignant lymphoma, acute leukemia, lymphosarcoma cell leukemia, B-cell leukemias, myelodysplastic syndromes, solid phase cancer, herpes viral infections, and/
  • the disease or disorder is a cancer, an infectious disease, or an autoimmune disorder.
  • the disease or disorder is a cancer.
  • the cancer is metastatic melanoma, a solid tumor, bladder cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma, hepatic metastasis of colonic origin, papillary thyroid carcinoma, acute myeloid leukemia, or asymptomatic myeloma.
  • the disease or disorder is an infectious disease.
  • the infectious disease is human immunodeficiency virus (HIV), chronic hepatitis C, cytomegalovirus, or hantavirus.
  • HIV human immunodeficiency virus
  • chronic hepatitis C chronic hepatitis C
  • cytomegalovirus cytomegalovirus
  • hantavirus hantavirus
  • the disease or disorder is an autoimmune disorder.
  • the autoimmune disorder is Chrohn’s disease, multiple sclerosis, systemic sclerosis, ocular myasthenia gravis, psoriasis or rheumatoid arthritis.
  • the autoimmune disorder is Chrohn’s disease, multiple sclerosis, systemic sclerosis, ocular myasthenia gravis, psoriasis or rheumatoid arthritis.
  • any of the antibodies or antigen binding fragments thereof can be used in generating a nucleic acid molecule comprising all or a portion of the sequence of the oligonucleotide or a complement thereof.
  • the antibody or antigen binding fragment thereof can be used in a method of associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
  • any of the antibodies or antigen binding fragments thereof can be used in the construction of a protein library.
  • the construction of a protein library comprises sequencing.
  • the construction of a protein library comprises the use of flow cytometry.
  • a method of detecting TMPRSS2 comprising a) contacting a sample with the antibody or antigen binding fragment thereof of any of the antibodies or antigen binding fragments thereof under conditions to bind said antibody or antigen binding fragment thereof to a TMPRSS2 receptor on said sample, wherein the binding generates the production of a receptor/antibody or antigen binding fragment thereof of complex; b) detecting the presence of the receptor/antibody or antigen binding fragment thereof of complexes; c) wherein the detecting comprises the presence or absence of the TMPRSS2 receptor on said sample.
  • a method of treating or preventing a disease or disorder associated with TMPRSS2 in a subject comprising: a) contacting a sample known or suspected to contain TMPRSS2 with the antibody or antigen binding fragment thereof any of the antibodies or antigen binding fragments thereof, b) detecting the presence of complexes comprising TMPRSS2 and the antibody or antigen binding fragment thereof; wherein the presence of the complexes indicates the presence of a disease or disorder; and c) administering to the subject the antibody or antigen binding fragment thereof of any of the antibodies or antigen binding fragments thereof
  • a method of diagnosing a disease or disorder comprising: a) isolating a sample from a subject, b) incubating the sample with the antibody or antigen binding fragment thereof of any of any of the antibodies or antigen binding fragments thereof, for a period of time sufficient to generate TMPRSS2:anti- TMPRSS2 complexes; c) detecting the presence or absence of the TMPRSS2:anti-TMPRSS2 complexes from the isolated tissue, and d) associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
  • the increase of TMPRSS2 over a control level in the location of interest of the tissue sample is indicative of a disease or disorder in a subject.
  • the detection comprises hybridization of a detectable moiety to the antibody or antigen binding fragment thereof.
  • the sample is contacted with a second antibody.
  • the second antibody is an antibody comprising a detectable moiety.
  • the detectable moiety comprises an oligonucleotide.
  • the detectable moiety comprises a fluorescent label.
  • the measurement comprises sequencing.
  • the detectable moiety comprises immunofluorescence.
  • the sample is a formalin-fixed paraffin- embedded sample.
  • the sample comprises a cell.
  • the sample comprises a tissue sample.
  • kits for example, a packaged combination of reagents in predetermined amounts with instructions for use (e.g., instructions for performing a diagnostic assay; instructions for performing a laboratory assay).
  • the kit is a diagnostic kit configured to detect in a sample (e.g., a biological sample).
  • the kit may include an identical isotype negative control irrelevant antibody to control for non-specific binding of the antibody.
  • the kit may include substrates and cofactors required by the enzyme (e.g., substrate precursor which provides the detectable chromophore or fluorophore).
  • reagents may be included such as stabilizers, buffers (e.g., a block buffer or lysis buffer), and the like.
  • the relative amounts of the various reagents may be varied widely to provide for concentrations in solution of the reagents that substantially optimize the sensitivity of the assay.
  • reagents may be provided as dry powders (e.g., lyophilized powder), including excipients that on dissolution will provide a reagent solution having the appropriate concentration.
  • CDRH1 heavy chain complementary determining region 1
  • Xi is S, N or D
  • X2 is Y or S
  • X3 is G, W or D
  • X4 is V, M or I
  • X5 is S, Q or N;
  • a heavy chain complementary determining region 2 comprising the sequence X1IX2X3X4X5X6X7X8X9X10X11X12X13X14X15X16 (SEQ ID NO: 82), wherein Xi is V, A, Y or W; X2 is W,Y,S or N; X3 is G, P, S or T; X 4 is D, V, G or Y; X 5 is G, D or T; X 6 is S or G; X 7 is T, D, R, S or E; Xs is N, T, I or P; X9 is Y, R, K or T; X10 is H, Y or F; Xn is S, T, P, N or A; X12 is A, Q, D or E; X13 is L, K, T or G; Xi4 is I, F or V; X15 is S or K; and Xi6 is G or no amino acid; and
  • a heavy chain complementary determining region 3 comprising the sequence X1X2X3X4X5X6X7X8X9X10X11X12X13 (SEQ ID NO: 83), wherein Xi is P, L, D, A , S or I; X2 is G, S, F, Y or H; X3 is N, P, D, Y or G; X 4 is N, S, Y, R, D or G; X 5 is Y, N F or no amino acid; Xe is D, Y S or no amino acid; X7 is W, D, Y, H or no amino acid; Xs is Y, F, G, A, W or no amino acid; X9 is F, D, A, M, Y or no amino acid; X10 is D, C or no amino acid; X11 is V, F, D or no amino acid; X12 is D, V or no amino acid; and X13 is Y or no
  • a light chain complementary determining region 1 comprising the sequence X1ASX2X3IX4X5X6X7X8 (SEQ ID NO: 84) wherein Xi is K or R; X2 is Q or E; X3 is D, S, E or N; X 4 is N, G, S or Y; X 5 is K, T, S or V; X 6 is Y or N; X7 is I, M or L; and Xs is A, H, S or T; and
  • a light chain complementary determining region 2 comprising the sequence X1X2X3X4X5X6X7 (SEQ ID NO: 85) wherein Xi is Y, R or A; X2 is T, A or G; X3 is S, N or T; X4 is T, E, R or N; X5 is L or S; Xe is Q, I, E, D or A; and X7 is P, S or D; and (iii) a light chain complementary determining region 3 (CDRL3) comprising the sequence X1X2X3X4X5X6X7X8X9 (SEQ ID NO: 86), wherein Xi is L or Q; X2 is Q or H; X3 is Y, S or F; X 4 is A, Y, D, H or W; X 5 is N, E, S or G; X 6 is L, W, P, Y or T; X7 is L or P; Xs
  • the antibody or antigen binding fragment thereof of embodiment 1, wherein the immunoglobulin heavy chain variable domain comprises: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,
  • the antibody or antigen binding fragment thereof of embodiment 1 or embodiment 2, wherein the immunoglobulin heavy chain variable domain comprises: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56; and a CDRH3 comprising the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63.
  • the immunoglobulin light chain variable domain comprises: a CDRL1 comprising the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,
  • the immunoglobulin light chain variable domain comprises: a CDRL1 comprising a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74; and a CDRL3 comprising a sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 45, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 51, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 57, or a sequence of amino acids that exhibits at least
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 46, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46;
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:52;
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 47, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47;
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53;
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 48, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48;
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:54;
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%,
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 45, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45;
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51;
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 61 or a sequence of amino acids that exhibits at least 80%
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 49, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49;
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:55;
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 62 or a sequence of amino acids that exhibits at least 80%
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:50;
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56;
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%
  • the CDRL1 comprises the sequence set forth in SEQ ID NO: 64, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64;
  • the CDRL2 comprises the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70;
  • the CDRL3 comprises the sequence set forth in SEQ ID NO: 75 or a sequence of amino acids that exhibits at least 80%, 81%
  • CDRL1 comprises the sequence set forth in SEQ ID NO: 64
  • CDRL2 comprises the sequence set forth in SEQ ID NO: 70
  • CDRL3 comprises the sequence set forth in SEQ ID NO: 75.
  • the CDRL1 comprises the sequence set forth in SEQ ID NO: 65 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:65
  • the CDRL2 comprises the sequence set forth in SEQ ID NO: 71 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:71
  • the CDRL3 comprises the sequence set forth in SEQ ID NO: 76 or a sequence of amino acids that exhibits at least 80%, 8
  • CDRL1 comprises the sequence set forth in SEQ ID NO: 65
  • CDRL2 comprises the sequence set forth in SEQ ID NO: 71
  • CDRL3 comprises the sequence set forth in SEQ ID NO: 76.
  • the CDRL1 comprises the sequence set forth in SEQ ID NO: 66 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:66
  • the CDRL2 comprises the sequence set forth in SEQ ID NO: 72 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:72
  • the CDRL3 comprises the sequence set forth in SEQ ID NO: 77 or a sequence of amino acids that exhibits at least 80%, 8
  • CDRL1 comprises the sequence set forth in SEQ ID NO: 66
  • CDRL2 comprises the sequence set forth in SEQ ID NO: 72
  • CDRL3 comprises the sequence set forth in SEQ ID NO: 77.
  • the CDRL1 comprises the sequence set forth in SEQ ID NO: 67 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:67
  • the CDRL2 comprises the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73
  • the CDRL3 comprises the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%
  • CDRL1 comprises the sequence set forth in SEQ ID NO: 67
  • CDRL2 comprises the sequence set forth in SEQ ID NO: 73
  • CDRL3 comprises the sequence set forth in SEQ ID NO: 78.
  • the CDRL1 comprises the sequence set forth in SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64
  • the CDRL2 comprises the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70
  • the CDRL3 comprises the sequence set forth in SEQ ID NO: 79 or a sequence of amino acids that exhibits at least 80%, 81%
  • CDRL1 comprises the sequence set forth in SEQ ID NO: 64
  • CDRL2 comprises the sequence set forth in SEQ ID NO: 70
  • CDRL3 comprises the sequence set forth in SEQ ID NO: 79.
  • the CDRL1 comprises the sequence set forth in SEQ ID NO: 68 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:68
  • the CDRL2 comprises the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73
  • the CDRL3 comprises the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%
  • the CDRL1 comprises the sequence set forth in SEQ ID NO: 69 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:69
  • the CDRL2 comprises the sequence set forth in SEQ ID NO: 74 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:74
  • the CDRL3 comprises the sequence set forth in SEQ ID NO: 80 or a sequence of amino acids that exhibits at least 80%, 8
  • the CDRH1 comprises the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50;
  • the CDRH2 comprises a sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55, or 56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50
  • the CDRH1 comprises the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50
  • the CDRH2 comprises the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55, or 56
  • the CDRH3 comprises the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63
  • the CDRL1 comprises the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69
  • the CDRL2 comprises the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74
  • the CDRL3 comprises the sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 57 or a sequence of amino acids that exhibits at least 80%
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 46 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46;
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:52;
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%,
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 47 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 59 or a sequence of amino acids that exhibits at least 80%
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 48 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:54
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%, 8
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 61 or a sequence of amino acids that exhibits at least 80%
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 49 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:55
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 62 or a sequence of amino acids that exhibits at least 80%
  • the CDRH1 comprises the sequence set forth in SEQ ID NO: 50 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:50
  • the CDRH2 comprises the sequence set forth in SEQ ID NO: 56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56
  • the CDRH3 comprises the sequence set forth in SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 20-26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20-26.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 20 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:20.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:21.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 22, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:22.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 24, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:24.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:25.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 27-33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27-33.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 28.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 29.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 30.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 31.
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 33.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 20-26 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20-26
  • the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 27-33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27-33.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 20, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 20; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 21; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NOS: 28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:28.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 22, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 22; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:29.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 23, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 23; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:30.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 24, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 24; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:31.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 25; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 32, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:32.
  • the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 26; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:33.
  • TMPRSS2 for use in the detection of TMPRSS2 in a sample.
  • 118 The antibody or antigen binding fragment thereof of embodiment 116 or embodiment 117, wherein the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs).
  • pDCs plasmacytoid dendritic cells
  • lymphocytes lymphocytes
  • leukocytes T cells
  • monocytes macrophages
  • neutrophils neutrophils
  • mDCs myeloid dendritic cells
  • innate lymphoid cells mast cells
  • eosinophils basophils
  • natural killer cells natural killer cells
  • PBMCs peripheral blood mononuclear cells
  • the detection comprises the use of a single antibody or antigen binding fragment thereof to bind a portion of TMPRSS2.
  • the detection comprises the use of two antibody or antigen binding fragments thereof, each capable of binding to a different portion of TMPRSS2.
  • TMPRSS2 is on the surface of a cell.
  • TMPRSS2 wherein the detection of TMPRSS2 is intracellular.
  • TMPRSS2 indicates the presence or absence of a disease or disorder.
  • a diagnostic antibody or antigen binding fragment thereof comprising the antibody or antigen binding fragment thereof of any of embodiments 1-133.
  • a kit comprising the antibody or antigen binding fragment thereof of any one of embodiments 1-133 or the diagnostic antibody or antigen binding fragment thereof of embodiment 134.
  • kits of embodiment 135, comprising a diagnostics kit configured to detect TMPRSS2 in a biological sample.
  • composition comprising the antibody or antigen binding fragment thereof of any of embodiments 1-133, and a pharmaceutically acceptable excipient.
  • composition of embodiment 137, wherein the antibody or antigen binding fragment thereof of is used as an adjuvant or in conjunction with an adjuvant.
  • An isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of the antibody or antigen binding fragment thereof of any of embodiments 1-133.
  • An isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain and the immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any of embodiments 1-133.
  • nucleic acid of any of embodiments 139-157 wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in any of SEQ ID NOs: 6-12; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in any of SEQ ID NOs: 13-19.
  • nucleic acid of embodiment 158 wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 6; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 13.
  • nucleic acid of embodiment 158 wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 7; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 14.
  • nucleic acid of embodiment 158 wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 8; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 15.
  • nucleic acid of embodiment 158 wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 9; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 16.
  • nucleic acid of embodiment 158 wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 10; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 17.
  • nucleic acid of embodiment 158 wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 12; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 19.
  • a recombinant expression vector comprising the isolated nucleic acid of any of embodiments 139- 165.
  • 167 A recombinant expression vector comprising a first expression cassette and a second expression cassette, wherein the first expression cassette comprises a nucleic acid molecule comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of any one of embodiments 1-133, and the second expression cassette comprises a nucleic acid molecule comprising a nucleotide sequence that encodes an immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any one of embodiments 1-133.
  • a recombinant expression vector comprising a first expression cassette and a second expression cassette, wherein the first expression cassette comprises a nucleic acid molecule comprising the nucleotide sequence of any of embodiments 139-165, and the second expression cassette comprises a nucleic acid molecule comprising the nucleotide sequence of any of embodiments 139-165.
  • An agent-drag conjugate comprising antibody or antigen binding fragment thereof of any of embodiments 1 -133.
  • a composition comprising the antibody-drag conjugate of embodiment 171, and a pharmaceutically acceptable carrier.
  • a method of detecting TMPRSS2 comprising a) contacting a sample with the antibody or antigen binding fragment thereof of any of embodiments 1-133, under conditions to bind said antibody or antigen binding fragment thereof to a TMPRSS2 receptor on said sample, wherein the binding generates the production of a receptor/antibody or antigen binding fragment thereof of complex; b) detecting the presence of the receptor/antibody or antigen binding fragment thereof of complexes; c) wherein the detecting comprises the presence or absence of the TMPRSS2 receptor on said sample.
  • a method of treating or preventing a disease or disorder associated with TMPRSS2 in a subject comprising: a) contacting a sample known or suspected to contain TMPRSS2 with the antibody or antigen binding fragment thereof of any of embodiments 1-133, b) detecting the presence of complexes comprising TMPRSS2 and the antibody or antigen binding fragment thereof; wherein the presence of the complexes indicates the presence of a disease or disorder; and c) administering to the subject the antibody or antigen binding fragment thereof of any of embodiments 1-133.
  • a method of diagnosing a disease or disorder comprising: a) isolating a sample from a subject b) incubating the sample with the antibody or antigen binding fragment thereof of any of embodiments 1-133, for a period of time sufficient to generate TMPRSS2:anti- TMPRSS2 complexes; c) detecting the presence or absence of the TMPRSS2:anti-TMPRSS2 complexes from the isolated tissue, and d) associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
  • sample is a formalin-fixed paraffin-embedded sample.
  • the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs).
  • pDCs plasmacytoid dendritic cells
  • lymphocytes lymphocytes
  • leukocytes T cells
  • monocytes macrophages
  • neutrophils neutrophils
  • mDCs myeloid dendritic cells
  • innate lymphoid cells mast cells
  • eosinophils basophils
  • natural killer cells natural killer cells
  • PBMCs peripheral blood mononuclear cells
  • invention 193 or embodiment 194, wherein the disease or disorder is chosen from non-viral cancers, virus-associated cancers, cancers associated with HBV infection, cancers associated with Epstein-Barr virus (EBV) infection, cancers associated with polyomavirus infection, erythema nodosum leprosum (ENL), autoimmune diseases, autoimmune inflammation, autoimmune thyroid diseases, B-cell lymphoma, T-cell lymphoma, acute myeloid leukemia, Hodgkin's Disease, acute myelogenous leukemia, acute myelomonocytic leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, B cell large cell lymphoma, malignant lymphoma, acute leukemia, lymphosarcoma cell leukemia, B-cell leukemias, myelodysplastic syndromes, solid phase cancer, herpes viral infections, and/or rejection of transplanted
  • EXAMPLE 1 Generation anti-TMPRSS2 antibody expressing hybridomas.
  • This Example describes the generation and characterization of hybridomas that secrete monoclonal antibodies that react with TMPRSS2 using murine models.
  • mice were immunized with a TMPRSS2 immunogen, and hybridomas were formed using standard protocols to fuse myeloma cells with spleens, and lymph node cells were drained and harvested. Successful fusions were selected into HAT medium, and cloned into approximately one cell per well in microtiter plates, after which culture supernatants were tested against TMPRSS2-expressing cell transfectants by flow cytometry. Wells were selected by assessment of staining profiles and then sub-cultured into larger vessels and subcloned. Hybridoma sub-clones were further characterized by flow cytometry using TMPRSS2-transfected cells.
  • Candidate clones expressing exemplary anti-TMPRSS2 antibodies were selected and screened using various methods, including by flow cytometry against human blood cells divided into distinct subsets (e.g., lymphocytes, monocytes, and the like), and against one or more cell lines generated from diseased and/or infected human cells. The percentage of positive cells in each blood cell subset was quantified as compared to isotype control.
  • distinct subsets e.g., lymphocytes, monocytes, and the like
  • EXAMPLE 2 Sequencing of exemplary anti-TMPRSS2 antibody variable regions.
  • CDRs and Framework regions Amino acid sequences of the individual variable domains (CDRs and Framework regions), including the CDR1, CDR2, and CDR3 regions, for both the heavy and light chains for seven different antibodies (clones), designated AB 1-7 (also referred to herein as antibodies 1-7, and clones 1-7), are shown in FIGs. 1 A and IB.
  • the various heavy and light chain CDR sequences are shown in Table El, below.
  • EXAMPLE 3 Detection of TMPRSS2 expressing cells using exemplary anti-TMPRSS2 antibodies.
  • This Example describes the ability of exemplary generated anti-TMPRSS2 antibodies to detect cells expressing TMPRSS2 by flow cytometry and immunohi stochemi stry .
  • exemplary anti-TMPRSS2 antibodies were assessed on cells from an immortalized human colorectal adenocarcinoma cell line (Caco-2 cells; (ATCC® HTB-37).
  • Caco-2 cells were grown in DMEM media supplemented with 10% FBS in T75 culture flask, to about 80% confluency. Once cells reached 80% confluency, cells where dislodged from the flask using accutase and suspended in cell staining buffer. 2pg of exemplary generated anti-TMPRSS2 antibodies were added, and allowed to incubate for 15 minutes. Cells were then washed twice with FACS wash buffer and stained with anti-mouse IgG-PE secondary antibody for 15 minutes. Cells where washed with FACS buffer and analyzed on a BD LSRII flow cytometer. A commercially available antibody was used as positive control. As shown in FIGs.
  • FIGs. 2A-2E exemplary tested anti-TMPRSS2 antibodies AB1, AB2, AB4, and AB6 (FIGs. 2A-2D) demonstrated similar staining profiles to a commercial antibody (CA; FIG. 2E), compared to isotype control, on Caco-2 cells.
  • Exemplary antibodies were further assessed on non-TMPRSS2 expressing HELA cells (ATCC® CCL-2) and white blood cells (lymphocytes, monocytes and granulocytes) isolated from healthy volunteer donors. Briefly, HeLa cells were grown in DMEM media supplemented with 10% FBS in T75 culture flask, to about 80% confluency. White blood cells were stained in whole blood followed by red blood cell lysis. Lysed blood was washed twice with FACS wash buffer and stained with anti-mouse IgG-PE secondary antibody for 15 minutes. Cells where washed with FACS buffer and analyzed on a BD LSRII flow cytometer. As shown in FIG. 3 A-3F and FIGs. 4A-4C, exemplary tested antibodies did not show surface staining on HELA cells (FIG. 3 A-3F), lymphocytes (FIG. 4A), monocytes (FIG. 4B), or granulocytes (FIG. 4C).
  • FFPE formalin-fixed paraffin-embedded
  • Samples were stained with 5pg/ml of purified exemplary anti-human TMPRSS2 antibody AB1, AB2, AB3, AB5 or AB7 over night at 4C. Cells were then washed and stained with 2.5ug/ml of Alexa-555 conjugated anti-mouse IgG for 1 hour at room temperature in the dark, followed by two washes in PBS. Samples were mounted using Antifade gold with DAPI and imaged using with Metamorph software and analyzed using image J.
  • exemplary generated anti-TMPRSS2 antibodies AB1, AB2, AB3, AB5 and AB7 are capable of staining TMPRSS2 expressing human kidney paraffin sections.
  • exemplary tested antibodies AB1, AB2, AB5 and AB7 did not result in staining on non-TMPRSS2 expressing lymph node sections.
  • EXAMPLE 4 Assessment of antibody blocking ability of exemplary anti-TMPRSS2 antibodies.
  • This Example describes the ability of exemplary generated anti-TMPRSS2 antibodies to block binding of other anti-TMPRSS2 antibodies.
  • Percentage original MFI was calculated by dividing the MFI of samples blocked with AB1, AB2, AB4 or AB6 by the MFI of samples blocked with the corresponding isotype control, as shown in Table E2. This value was subtracted from 100 to get a blocking percentage. The formula is shown below: n / . . chorus > > , [MFI blocking] .
  • exemplary anti-TMPRSS2 antibodies AB1, AB2, AB4 and AB6 are capable of blocking binding of an exemplary tested commercial antibody. These results suggest that exemplary anti-TMPRSS2 antibodies AB1, AB2, AB4 and AB6 are capable of recognizing similar epitopes.
  • This Example describes the functional assessment of exemplary anti-TMPRSS2 antibodies as measured by the inhibition of TMPRSS2 protease activity and their effect on cell migration, as compared to both commercially available antibodies and the broad spectrum serine protease inhibitor Nafamostat, which has been shown to inhibit TMPRSS2 activity.
  • Boc-Gln-Ala-ArgAMC a protease substrate that exhibits fluorescence when cleaved by proteases, was added at a final concentration of 5pM. Fluorescence was measured after 3h at 37C, and protease activity was normalized to untreated controls (platelets and substrate only). As shown in Fig. 6A, AB1 showed inhibition of protease activity at multiple concentrations tested compared to control.
  • recombinant TMPRSS2 was incubated with AB 1 diluted at a final concentration of 2, 1, 0.5, 0.25, 0.125 and 0.6 pg per well.
  • the broad protease inhibitor Nafamostat Mesylate at final concentrations of 50pM, 25 pM, 12 pM and 6 pM and untreated platelets (platelets and substrate only) and untreated recombinant TMPRSS2 used as controls.
  • AB1 showed inhibition of protease activity at multiple concentrations tested compared to Nafamostat and control.
  • AB1 inhibited the ability of Caco-2 cells to invade the Matrigel and migrate to the bottom of the membrane to a higher extent than any commercially tested antibody, and to a degree similar to a broad spectrum serine protease inhibitor (Nafamostat).
  • anti-TMPRSS2 antibody AB1 is capable of inhibiting the functional activity of TMPRSS2 with stronger blocking activity than any tested commercially available antibody.
  • degree of inhibition was comparable to the potent inhibitory effects of the serine protease inhibitor Nafamostat.
  • Nafamostat has additional inhibitory effects on multiple types of serine proteases given it’s broad spectrum activity
  • anti-TMPRSS2 antibodies have the potential for more specific activity and less off target effects, providing potential advantages when used for TMPRSS2 specific purposes.

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Abstract

Compositions and methods comprising TMPRSS2 binding antibodies and antigen binding fragments thereof are provided.

Description

TMPRSS2 BINDING ANTIBODIES AND ANTIGEN BINDING FRAGMENTS
THEREOF
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims priority to U.S. Provisional Patent Application No. 63/297,492, filed on January 7, 2022, and U.S. Provisional Patent Application No. 63/416,370, filed on October 14, 2022, the disclosures of which are incorporated herein by reference in their entirety for all purposes.
FIELD OF THE INVENTION
[0002] The present disclosure relates, in some aspects, to antibodies or antigen-binding fragments thereof that bind TMPRSS2, as well as methods, systems and kits for detection of TMPRSS2. In certain aspects, the present disclosure relates to antibodies or antigen binding fragments thereof for use in determining levels of TMPRSS2 in a sample containing or suspected of containing TMPRSS2. In some aspects, the present disclosure relates to antibodies or antigen-binding fragments thereof for use in diagnosing or treating an individual with or suspected of having a disease or disorder associated with TMPRSS2.
BACKGROUND
[0003] The transmembrane protease serine 2 (TMPRSS2), is a plasma membrane anchored serine protease that has been associated with both physiological and pathological processes including digestion, tissue remodeling, blood coagulation, fertility, inflammatory responses, tumor cell invasion, apoptosis and pain. The function of TMPRSS2 has also been implicated in facilitating viral pathogenesis for the influenza A virus and human coronaviruses SARS- CoV and SARS-Cov-2. However, currently available anti-TMPRSS2 antibodies and associated methods are limited in their range of both in vitro and in vivo applications. Thus, there remains a need for reagents, devices and methods of detecting TMPRSS2, and diagnosing and/or treating TMPRSS2 related diseases or disorders. Provided herein are embodiments that meet such needs.
BRIEF SUMMARY
[0004] Provided herein are antibodies, including antigen-binding fragments thereof, that bind all or a portion thereof of TMPRSS2, compositions containing such antibodies or antigen-binding fragments thereof, combinations of such antibodies or antigen-binding fragments thereof and methods of use. In particular embodiments, the antibodies or antigenbinding fragments thereof are used in methods of detecting the presence of TMPRSS2 through imaging, including molecular, medical and diagnostic imaging.
[0005] Provided herein are antibodies or antigen-binding fragments thereof, including those that specifically bind to a TMPRSS2, such as a human TMPRSS2, wherein the antibodies or antigen-binding fragments contain particular complementarity determining regions (CDRs), including heavy chain CDRs (i.e., CDRH1, CDRH2, and/or CDRH3) and light chain CDRs (i.e., CDRL1, CDRL2, and/or CDRL3), such as any described herein. In some embodiments, the antibody or antigen-binding fragment thereof includes a heavy chain variable domain and a light chain variable domain, such as any described herein.
[0006] Provided herein is an isolated antibody or antigen binding fragment thereof that binds TMPRSS2 or a portion thereof, comprising a) an immunoglobulin heavy chain variable domain comprisingb (i) a heavy chain complementary determining region 1 (CDRH1) comprising the sequence X1X2X3X4X5 (SEQ ID NO: 81), wherein Xi is S, N or D; X2 is Y or S; X3 is G, W or D; X4 is V, M or I; and X5 is S, Q or N; and (ii) a heavy chain complementary determining region 2 (CDRH2) comprising the sequence
X1IX2X3X4X5X6X7X8X9X10X11X12X13X14X15X16 (SEQ ID NO: 82), wherein Xi is V, A, Y or W; X2 is W,Y,S or N; X3 is G, P, S or T; X4 is D, V, G or Y; X5 is G, D or T; X6 is S or G; X7 is T, D, R, S or E; Xs is N, T, I or P; X9 is Y, R, K or T; X10 is H, Y or F; Xu is S, T, P, N or A; X12 is A, Q, D or E; X13 is L, K, T or G; X14 is I, F or V; X15 is S or K; and Xi6 is G or no amino acid; and (iii) a heavy chain complementary determining region 3 (CDRH3) comprising the sequence X1X2X3X4X5X6X7X8X9X10X11X12X13 (SEQ ID NO: 83), wherein Xi is P, L, D, A , S or I; X2 is G, S, F, Y or H; X3 is N, P, D, Y or G; X4 is N, S, Y, R, D or G; X5 is Y, N F or no amino acid; Xe is D, Y S or no amino acid; X7 is W, D, Y, H or no amino acid; Xs is Y, F, G, A, W or no amino acid; X9 is F, D, A, M, Y or no amino acid; X10 is D, C or no amino acid; Xu is V, F, D or no amino acid; X12 is D, V or no amino acid; and X13 is Y or no amino acid; and b) an immunoglobulin light chain variable domain comprising (i) a light chain complementary determining region 1 (CDRL1) comprising the sequence X1ASX2X3IX4X5X6X7X8 (SEQ ID NO: 84) wherein Xi is K or R; X2 is Q or E; X3 is D, S, E or N; X4 is N, G, S or Y; X5 is K, T, S or V; X6 is Y or N; X7 is I, M or L; and Xs is A, H, S or T; and (ii) a light chain complementary determining region 2 (CDRL2) comprising the sequence X1X2X3X4X5X6X7 (SEQ ID NO: 85) wherein Xi is Y, R or A; X2 is T, A or G; X3 is S, N or T; X4 is T, E, R or N; X5 is L or S; Xe is Q, I, E, D or A; and X7 is P, S or D; and (iii) a light chain complementary determining region 3 (CDRL3) comprising the sequence X1X2X3X4X5X6X7X8X9 (SEQ ID NO: 86), wherein Xi is L or Q; X2 is Q or H; X3 is Y, S or F; X4 is A, Y, D, H or W; X5 is N, E, S or G; X6 is L, W, P, Y or T; X7 is L or P; Xs is T, L or Y; and X9 is T or no amino acid.
[0007] In some embodiments, the immunoglobulin heavy chain variable domain includes: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 51, 52, 53, 54, 55 or 56; and a CDRH3 comprising the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63.
[0008] In some embodiments, the immunoglobulin heavy chain variable domain includes: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56; and a CDRH3 comprising the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63.
[0009] In some embodiments, the immunoglobulin light chain variable domain includes: a CDRL1 comprising the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 70, 71, 72, 73 or 74; and a CDRL3 comprising the sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 75, 76, 77, 78, 79 or 80. In some embodiments, the immunoglobulin light chain variable domain includes: a CDRL1 comprising a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74; and a CDRL3 comprising a sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
[0010] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 45, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45, the CDRH2 includes the sequence set forth in SEQ ID NO: 51, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; and the CDRH3 includes the sequence set forth in SEQ ID NO: 57, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:57. In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 45, the CDRH2 includes the sequence set forth in SEQ ID NO: 51; and the CDRH3 includes the sequence set forth in SEQ ID NO: 57.
[0011] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 46, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46; the CDRH2 includes the sequence set forth in SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 52; and the CDRH3 includes the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:58. In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 46, the CDRH2 includes the sequence set forth in SEQ ID NO: 52; and the CDRH3 includes the sequence set forth in SEQ ID NO: 58.
[0012] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 47, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47; the CDRH2 includes the sequence set forth in SEQ ID NO: 53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53; and the CDRH3 includes the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:58.
[0013] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 47, the CDRH2 includes the sequence set forth in SEQ ID NO: 53; and the CDRH3 includes the sequence set forth in SEQ ID NO: 58.
[0014] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 48, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48; the CDRH2 includes the sequence set forth in SEQ ID NO: 54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 54; and the CDRH3 includes the sequence set forth in SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:60.
[0015] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 48, the CDRH2 includes the sequence set forth in SEQ ID NO: 54; and the CDRH3 includes the sequence set forth in SEQ ID NO: 60.
[0016] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 45, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45; the CDRH2 includes the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; and the CDRH3 includes the sequence set forth in SEQ ID NO: 61 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:61.
[0017] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 45, the CDRH2 includes the sequence set forth in SEQ ID NO: 51; and the CDRH3 includes the sequence set forth in SEQ ID NO: 61.
[0018] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 49, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49; the CDRH2 includes the sequence set forth in SEQ ID NO: 55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 55; and the CDRH3 includes the sequence set forth in SEQ ID NO: 62 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:62.
[0019] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 49, the CDRH2 includes the sequence set forth in SEQ ID NO: 55; and the CDRH3 includes the sequence set forth in SEQ ID NO: 62.
[0020] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 50; the CDRH2 includes the sequence set forth in SEQ ID NO: 56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56; and the CDRH3 includes the sequence set forth in SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:63.
[0021] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 50, the CDRH2 includes the sequence set forth in SEQ ID NO: 56; and the CDRH3 includes the sequence set forth in SEQ ID NO: 63.
[0022] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 64, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64; the CDRL2 includes the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and the CDRL3 includes the sequence set forth in SEQ ID NO: 75 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:75.
[0023] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 64, the CDRL2 includes the sequence set forth in SEQ ID NO: 70; and the CDRL3 includes the sequence set forth in SEQ ID NO: 75.
[0024] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 65 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:65, the CDRL2 includes the sequence set forth in SEQ ID NO: 71 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:71; and the CDRL3 includes the sequence set forth in SEQ ID NO: 76 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 76. [0025] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 65, the CDRL2 includes the sequence set forth in SEQ ID NO: 71; and the CDRL3 includes the sequence set forth in SEQ ID NO: 76.
[0026] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 66 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 66, the CDRL2 includes the sequence set forth in SEQ ID NO: 72 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:72; and the CDRL3 includes the sequence set forth in SEQ ID NO: 77 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 77.
[0027] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 66, the CDRL2 includes the sequence set forth in SEQ ID NO: 72; and the CDRL3 includes the sequence set forth in SEQ ID NO: 77.
[0028] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 67 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 67, the CDRL2 includes the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and the CDRL3 includes the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 78.
[0029] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 67, the CDRL2 includes the sequence set forth in SEQ ID NO: 73; and the CDRL3 includes the sequence set forth in SEQ ID NO: 78.
[0030] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64, the CDRL2 includes the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and the CDRL3 includes the sequence set forth in SEQ ID NO: 79 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 79.
[0031] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 64, the CDRL2 includes the sequence set forth in SEQ ID NO: 70; and the CDRL3 includes the sequence set forth in SEQ ID NO: 79.
[0032] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 68 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 68, the CDRL2 includes the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and the CDRL3 includes the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 78.
[0033] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 68, the CDRL2 includes the sequence set forth in SEQ ID NO: 73; and the CDRL3 includes the sequence set forth in SEQ ID NO: 78.
[0034] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 69 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 69, the CDRL2 includes the sequence set forth in SEQ ID NO: 74 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 74; and the CDRL3 includes the sequence set forth in SEQ ID NO: 80 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 80.
[0035] In some embodiments, the CDRL1 includes the sequence set forth in SEQ ID NO: 69, the CDRL2 includes the sequence set forth in SEQ ID NO: 74; and the CDRL3 includes the sequence set forth in SEQ ID NO: 80.
[0036] In some embodiments, the CDRH1 includes the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; the CDRH2 includes a sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55, or
56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 51, 52, 53, 54, 55, or 56; the CDRH3 includes a sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:
57, 58, 59, 60, 61, 62 or 63; the CDRL1 includes a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69; the CDRL2 includes a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 70, 71, 72, 73 or 74; the CDRL3 includes a sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 75, 76, 77, 78, 79 or 80. In some embodiments, CDRH1 includes the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50; the CDRH2 includes the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55, or 56; the CDRH3 includes the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63; the CDRL1 includes the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69; the CDRL2 includes the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74; and the CDRL3 includes the sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
[0037] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45, the CDRH2 includes the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; the CDRH3 includes the sequence set forth in SEQ ID NO: 57 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:57; the CDRL1 includes the sequence set forth in SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64, the CDRL2 includes the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and the CDRL3 includes the sequence set forth in SEQ ID NO: 75 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:75. In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 45, the CDRH2 includes the sequence set forth in SEQ ID NO: 51; the CDRH3 includes the sequence set forth in SEQ ID NO: 57; the CDRL1 includes the sequence set forth in SEQ ID NO: 64, the CDRL2 includes the sequence set forth in SEQ ID NO: 70; and the CDRL3 includes the sequence set forth in SEQ ID NO: 75.
[0038] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 46 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46; the CDRH2 includes the sequence set forth in SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 52; the CDRH3 includes the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:58; the CDRL1 includes the sequence set forth in SEQ ID NO: 65 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:65, the CDRL2 includes the sequence set forth in SEQ ID NO: 71 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:71; and the CDRL3 includes the sequence set forth in SEQ ID NO: 76 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:76. In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 46, the CDRH2 includes the sequence set forth in SEQ ID NO: 52; the CDRH3 includes the sequence set forth in SEQ ID NO: 58; the CDRL1 includes the sequence set forth in SEQ ID NO: 65, the CDRL2 includes the sequence set forth in SEQ ID NO: 71; and the CDRL3 includes the sequence set forth in SEQ ID NO: 76.
[0039] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 47 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47, the CDRH2 includes the sequence set forth in SEQ ID NO: 53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53; the CDRH3 includes the sequence set forth in SEQ ID NO: 59 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:59; the CDRL1 includes the sequence set forth in SEQ ID NO: 66 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 66, the CDRL2 includes the sequence set forth in SEQ ID NO: 72 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:72; and the CDRL3 includes the sequence set forth in SEQ ID NO: 77 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:77. In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 47, the CDRH2 includes the sequence set forth in SEQ ID NO: 53; the CDRH3 includes the sequence set forth in SEQ ID NO: 59; the CDRL1 includes the sequence set forth in SEQ ID NO: 66, the CDRL2 includes the sequence set forth in SEQ ID NO: 72; and the CDRL3 includes the sequence set forth in SEQ ID NO: 77.
[0040] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 48 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48, the CDRH2 includes the sequence set forth in SEQ ID NO: 54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 54; the CDRH3 includes the sequence set forth in SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 60; the CDRL1 includes the sequence set forth in SEQ ID NO: 67 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 67, the CDRL2 includes the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and the CDRL3 includes the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:78. In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 48, the CDRH2 includes the sequence set forth in SEQ ID NO: 54; the CDRH3 includes the sequence set forth in SEQ ID NO: 60; the CDRL1 includes the sequence set forth in SEQ ID NO: 67, the CDRL2 includes the sequence set forth in SEQ ID NO: 73; and the CDRL3 includes the sequence set forth in SEQ ID NO: 78.
[0041] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO:
45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45, the CDRH2 includes the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; the CDRH3 includes the sequence set forth in SEQ ID NO: 61 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:61; the CDRL1 includes the sequence set forth in SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64, the CDRL2 includes the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and the CDRL3 includes the sequence set forth in SEQ ID NO: 79 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:79. In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 45, the CDRH2 includes the sequence set forth in SEQ ID NO: 51; the CDRH3 includes the sequence set forth in SEQ ID NO: 61; the CDRL1 includes the sequence set forth in SEQ ID NO: 64, the CDRL2 includes the sequence set forth in SEQ ID NO: 70; and the CDRL3 includes the sequence set forth in SEQ ID NO: 79.
[0042] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 49 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49, the CDRH2 includes the sequence set forth in SEQ ID NO: 55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 55; the CDRH3 includes the sequence set forth in SEQ ID NO: 62 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 62; the CDRL1 includes the sequence set forth in SEQ ID NO: 68 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 68, the CDRL2 includes the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and the CDRL3 includes the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:78. In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 49, the CDRH2 includes the sequence set forth in SEQ ID NO: 55; the CDRH3 includes the sequence set forth in SEQ ID NO: 62; the CDRL1 includes the sequence set forth in SEQ ID NO: 68, the CDRL2 includes the sequence set forth in SEQ ID NO: 73; and the CDRL3 includes the sequence set forth in SEQ ID NO: 78.
[0043] In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 50 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:50, the CDRH2 includes the sequence set forth in SEQ ID NO: 56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56; the CDRH3 includes the sequence set forth in SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 63; the CDRL1 includes the sequence set forth in SEQ ID NO: 69 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 69, the CDRL2 includes the sequence set forth in SEQ ID NO: 74 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 74; and the CDRL3 includes the sequence set forth in SEQ ID NO: 80 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:80. In some embodiments, the CDRH1 includes the sequence set forth in SEQ ID NO: 50, the CDRH2 includes the sequence set forth in SEQ ID NO: 56; the CDRH3 includes the sequence set forth in SEQ ID NO: 63; the CDRL1 includes the sequence set forth in SEQ ID NO: 69, the CDRL2 includes the sequence set forth in SEQ ID NO: 74; and the CDRL3 includes the sequence set forth in SEQ ID NO: 80.
[0044] In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 20-26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20-26. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 20-26.
[0045] In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 20 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:20. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 20. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:21. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 21. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 22, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:22. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 22. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 23, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:23. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 23. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 24, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:24. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 24. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:25. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 25. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:26. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 26.
[0046] In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 27-33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27-33. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 27-33. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 27. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 28. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 28. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 9. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 9. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 30. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 30. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 31.
[0047] In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 31. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 32, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 32. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 32. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 33. In some embodiments, the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 33.
[0048] In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 20-26 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20-26, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 27-33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27-33. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 20-26, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in any of SEQ ID NOS: 27-33.
[0049] In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 20, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 20; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 20, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 27. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 21; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NOS: 28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:28. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 21, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 28. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 22, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 22; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:29. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 22, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 29. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 23, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 23; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:30. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 23, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 30. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 24, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 24; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:31. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 24, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 31. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 25; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 32, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:32. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO:25, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO:32. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 26; and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:33. In some embodiments, the immunoglobulin heavy chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 26, and the immunoglobulin light chain variable domain includes the amino acid sequence set forth in SEQ ID NO: 33.
[0050] In some embodiments, any of the antibody or antigen binding fragment thereof includes one immunoglobulin heavy chain variable domain and one immunoglobulin light chain variable domain. In some embodiments, any of the antibody or antigen binding fragment thereof includes two immunoglobulin heavy chain variable domains and two immunoglobulin light chain variable domains.
[0051] In some of any of the embodiments herein, the antibody or antigen-binding fragment is isolated. In some of any of the embodiments herein, the antibody is a humanized antibody, a chimeric antibody or a human antibody. In some of any of the embodiments herein, the antibody is a murine antibody.
[0052] In some of any of the embodiments herein, the antibody is an antigen-binding fragment thereof. In a further embodiment, the antigen-binding fragment thereof is a Fab, Fab'- SH, Fv, scFv, or (Fab')2 fragment. In some of any of the embodiments herein, the antibody is a full length or intact antibody.
[0053] In some of any of the embodiments herein, the antibody further comprises a heavy chain constant domain and/or a light chain constant domain. In a further embodiment, the heavy chain and/or light constant domain is murine or human. In some further embodiments, the heavy chain constant domain is IgGl, IgG2a, IgG2b or IgM.
[0054] In some of any of the embodiments herein, the antibody is a monoclonal antibody.
[0055] In some of any of the embodiments herein, the antibody is attached to a label. In a further embodiment, the label is a fluorescent dye, a fluorescent protein, a radioisotope, a chromophores, a metal ion, gold particles, silver particles, magnetic particles, a polypeptides, an enzyme, streptavidin, biotin, a luminescent compound, or an oligonucleotide. In some embodiments, the oligonucleotide includes a sample barcode sequence. In some embodiments, the oligonucleotide includes a binding site for a primer and an anchor. [0056] In some embodiments, the detectable marker or label is conjugated directly to the antigen or antigen binding fragment thereof. In some embodiments, the detectable marker or label is conjugated to the oligonucleotide. In some embodiments, the antibody or antigen binding fragment thereof is non-diffusively immobilized on a solid support.
[0057] In another aspect, the disclosure provides an isolated antibody that specifically binds to, wherein the isolated antibody competes binding to the with an antibody described herein.
[0058] In some embodiments of the antibodies described herein, the antibody is a monoclonal antibody. In certain embodiments, the antibody is a humanized antibody. In certain embodiments, the antibody comprises one or more human framework regions.
[0059] In some aspects, provided herein is a combination of antibodies or antigen-binding fragments thereof, wherein the combination comprises two or more anti-TMPRSS2 antibodies or antigen-binding fragments described herein. In some embodiments, the two or more antibodies or antigen-binding fragments comprise one or more first antibody or antigenbinding fragment thereof that binds to a first epitope or region within TMPRSS2; and one or more second antibody or antigen-binding fragment thereof that binds to a second epitope or region within TMPRSS2. In some further embodiments, the one or more first antibody or antigen-binding fragments thereof, and the one or more second antibody or antigen-binding fragments thereof bind to a non-overlapping epitope or region of TMPRSS2 (e.g., human TMPRSS2) and/or do not compete for binding to TMPRSS2. Further, in some embodiments, the antibody is conjugated to a detectable marker or label. In some embodiments, the at least one of the antibodies or antigen-binding fragments of the combination of two or more anti- TMPRSS2 antibodies or antigen-binding fragments described herein, optionally the one or more first antibody or antigen-binding fragment thereof or the one or more second antibody or antigen-binding fragment thereof, is conjugated to a label. In some embodiments, the at least one of the antibodies or antigen-binding fragments, optionally the one or more first antibody or antigen-binding fragment thereof or the one or more second antibody or antigen-binding fragment thereof, is attached or immobilized to a solid support. In some embodiments, the one or more first or second antibody or antigen-binding fragment is attached or immobilized to a solid support and the other of the one or more first or second antibody or antigen-binding fragment is conjugated to a label. In some embodiments, the label is a fluorescent dye, a fluorescent protein, a radioisotope, a chromophore, a metal ion, gold particles, silver particles, magnetic particles, a polypeptide, an enzyme, streptavidin, biotin, a luminescent compound, or an oligonucleotide. In some embodiments, the solid support is a bead, a column, an array, an assay plate, a microwell, a stick, a filter, or a strip. In certain embodiments, the antibody is non- diffusively immobilized on a solid support. In a further embodiment, the device is a rapid detection device or a rapid diagnostic device.
[0060] In another aspect, the disclosure features an isolated nucleic acid encoding an isolated antibody described herein. The disclosure also provides an expression vector comprising the nucleic acid described herein. Further, the disclosure also provides an isolated host cell comprising the expression vector described herein.
[0061] In some embodiments, the antibody or antigen binding fragment thereof provided herein can be used in the detection of TMPRSS2 in a sample. In some embodiments, the antibody or antigen binding fragment thereof binds to a cell expressing TMPRSS2 in a sample. In some embodiments, the sample includes immune cells. In some embodiments, the sample includes a heterogenous population of immune cells. In some embodiments, the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs). In some embodiments, the sample includes a cell with a disease or disorder. In some embodiments, the disease or disorder is a cancer, an autoimmune disorder, an inflammatory disorder, a neurologic disorder, or an infection. In some embodiments, the cancer is acute myeloid leukemia, acute lymphoblastic leukemia, colorectal, ovarian, gynecologic, liver, glioblastoma, Hodgkin lymphoma, chronic lymphocytic leukemia, esophagus, gastric, pancreas, colon, kidney, head and neck, lung and melanoma. In some embodiments, the detection includes the use of a single antibody or antigen binding fragment thereof to bind a portion of TMPRSS2. In some embodiments, the detection includes the use of two antibody or antigen binding fragments thereof, each capable of binding to a different portion of TMPRSS2. In some embodiments, the detection of TMPRSS2 is on the surface of a cell. In some embodiments, the detection of TMPRSS2 is intracellular. In some embodiments, the detection of TMPRSS2 indicates the presence or absence of a disease or disorder. In some embodiments, the detection is performed in vitro. In some embodiments, the detection is performed in vivo. [0062] In some embodiments, the antibody or antigen binding fragment thereof binds to a TMPRSS2 expressing cell. In some embodiments, the binding to the TMPRSS2 expressing cell decreases the production of androgenic hormones. In some embodiments, the binding to the TMPRSS2 expressing cell inhibits proteolytic cleavage of ACE2 receptor. In some embodiments, the binding to the TMPRSS2 expressing cell inhibits or reduces cleavage of coronavirus spike glycoproteins. In some embodiments, the binding to the TMPRSS2 expressing cell inhibits or reduces viral uptake into a host cell.
[0063] Provided herein is a diagnostic antibody or antigen binding fragment thereof which includes any of the antibody or antigen binding fragment thereof described herein. Provided herein is a kit comprising the antibody or antigen binding fragment thereof of any one of embodiments described herein. In some embodiments, the kit is a diagnostics kit configured to detect TMPRSS2 in a biological sample.
[0064] Provided herein is a composition comprising the antibody or antigen binding fragment thereof of any of the embodiments described herein and a pharmaceutically acceptable excipient. In some embodiments, the antibody or antigen binding fragment thereof of is used as an adjuvant or in conjunction with an adjuvant.
[0065] Provided herein is an isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of the agent of any of embodiments described herein. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in any of SEQ ID NOs: 6-12. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 6. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 7. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 8. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 9. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 10. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 11. In some embodiments, the immunoglobulin heavy chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 12.
[0066] Provided herein is an isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin light chain variable domain of the agent of any of the embodiments described herein. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in any of SEQ ID NOs: 13-19. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 13. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 14. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 15. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 16. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 17. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 18. In some embodiments, the immunoglobulin light chain variable domain includes the sequence of nucleotides set forth in SEQ ID NO: 19.
[0067] Provided herein is an isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain and the immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any of the embodiments described herein. In some embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in any of SEQ ID NOs: 6-12; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in any of SEQ ID NOs: 13-19. In some embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 6; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 13. In some embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 7; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 14. In some embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 8; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 15. In some embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 9; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 16. In some embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 10; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 17. In some embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 11; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 18. In some embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain includes the sequence set forth in SEQ ID NO: 12; and the immunoglobulin light chain variable domain includes the sequence of amino acids set forth in SEQ ID NO: 19.
[0068] Provided herein is a recombinant expression vector comprising the isolated nucleic acid of any of the embodiments described herein. Provided herein is a recombinant expression vector comprising a first expression cassette and a second expression cassette, wherein the first expression cassette includes a nucleic acid molecule comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of any one of the embodiments described herein and the second expression cassette includes a nucleic acid molecule comprising a nucleotide sequence that encodes an immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any one of any of the embodiments described herein.
[0069] Provided herein is a recombinant expression vector comprising a first expression cassette and a second expression cassette, wherein the first expression cassette includes a nucleic acid molecule comprising the nucleotide sequence of any of the embodiments described herein, and the second expression cassette includes a nucleic acid molecule comprising the nucleotide sequence of any of the embodiments described herein. In some embodiments, the first and second expression cassettes include a promoter.
[0070] Provided herein is a host cell transfected with the recombinant expression vector of any of the embodiments described herein. [0071] Provided herein is an agent-drug conjugate comprising antibody or antigen binding fragment thereof of any of the embodiments described herein. Provided herein is a composition comprising the antibody-drug conjugate and a pharmaceutically acceptable carrier. Provided herein is a method of detecting TMPRSS2, comprising a) contacting a sample with the antibody or antigen binding fragment thereof of any of the embodiments described herein under conditions to bind said antibody or antigen binding fragment thereof to a TMPRSS2 receptor on said sample, wherein the binding generates the production of a receptor/antibody or antigen binding fragment thereof of complex; b) detecting the presence of the receptor/antibody or antigen binding fragment thereof of complexes; c) wherein the detecting includes the presence or absence of the TMPRSS2 receptor on said sample.
[0072] Provided herein is a method of treating or preventing a disease or disorder associated with TMPRSS2 in a subject, comprising: a) contacting a sample known or suspected to contain TMPRSS2 with the antibody or antigen binding fragment thereof of any of the embodiments described herein b) detecting the presence of complexes comprising TMPRSS2 and the antibody or antigen binding fragment thereof; wherein the presence of the complexes indicates the presence of a disease or disorder; and c) administering to the subject the antibody or antigen binding fragment thereof of any of any of the embodiments described herein.
[0073] Provided herein is a method of diagnosing a disease or disorder, comprising: a) isolating a sample from a subject b) incubating the sample with the antibody or antigen binding fragment thereof of any of any of the embodiments described herein, for a period of time sufficient to generate TMPRSS2:anti-TMPRSS2 complexes; c) detecting the presence or absence of the TMPRSS2:anti-TMPRSS2 complexes from the isolated tissue, and d) associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
[0074] In some embodiments, the increase of TMPRSS2 over a control level in the location of interest of the tissue sample is indicative of a disease or disorder in a subject.
[0075] In some embodiments, the method is performed in vitro. In some embodiments, the method is performed in vivo. In some embodiments, the detection includes intracellular detection. In some embodiments, the detection includes detection on the surface of a cell. In some embodiments, the detection includes hybridization of a detectable moiety to the antibody or antigen binding fragment thereof. In some embodiments, the sample is contacted with a second antibody. In some embodiments, the second antibody is an antibody comprising a detectable moiety. In some embodiments, the detectable moiety includes an oligonucleotide. In some embodiments, the detectable moiety includes a fluorescent label. In some embodiments, the measurement includes sequencing. In some embodiments, the detectable moiety includes immunofluorescence. In some embodiments, the sample is a formalin-fixed paraffin-embedded sample. In some embodiments, the sample includes a cell. In some embodiments, the sample includes a tissue sample.
[0076] In some embodiments, the sample includes immune cells. In some embodiments, the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs). In some embodiments, the sample includes a tissue or cells associated with a disease or disorder. In some embodiments, the disease or disorder is a cancer, an autoimmune disorder, an inflammatory disorder, or an infection. In some embodiments, the disease or disorder is chosen from non-viral cancers, virus-associated cancers, cancers associated with HBV infection, cancers associated with Epstein-Barr virus (EBV) infection, cancers associated with polyomavirus infection, erythema nodosum leprosum (ENL), autoimmune diseases, autoimmune inflammation, autoimmune thyroid diseases, B-cell lymphoma, T-cell lymphoma, acute myeloid leukemia, Hodgkin's Disease, acute myelogenous leukemia, acute myelomonocytic leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, B cell large cell lymphoma, malignant lymphoma, acute leukemia, lymphosarcoma cell leukemia, B-cell leukemias, myelodysplastic syndromes, solid phase cancer, herpes viral infections, and/or rejection of transplanted tissues or organs.
[0077] In some embodiments, the antibody or antigen binding fragment thereof can be used in a method of associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
[0078] In some embodiments, the antibody or antigen binding fragment thereof can be used in a method of detecting TMPRSS2 in a tissue sample. In some embodiments, the method includes generating a nucleic acid molecule comprising all or a portion of the sequence of the oligonucleotide or a complement thereof. [0079] In some embodiments, the antibody or antigen binding fragment thereof can be used in the construction of a protein library. In some embodiments, the construction of a protein library includes sequencing. In some embodiments, the construction of a protein library includes the use of flow cytometry.
BRIEF DESCRIPTION OF THE DRAWINGS
[0080] The drawings illustrate certain embodiments of the technology and are not limiting. For clarity and ease of illustration, the drawings are not made to scale and, in some instances, various aspects may be shown exaggerated or enlarged to facilitate an understanding of particular embodiments.
[0081] FIGs. 1A-1B show the amino acid sequences of the variable domains of the immunoglobulin heavy (SEQ ID NOS:20-26 and light (SEQ ID NOS: 27-33) chains of the seven different anti-TMPRSS2 antibodies (AB 1-7) provided herein. The CDR regions of each of the heavy and light chains are shown in bold and underline. In each of the alignments, three characters
Figure imgf000031_0001
and “.”) are used: indicates positions that have a single, fully conserved residue; indicates that one of the following “strong” residue groups is fully conserved: STA; NEQK; NHQK; NDEQ; QHRK; MILV; MILF; HY; and FYW; and indicates that one of the following “weaker” residue groups is fully conserved: CSA; ATV; SAG; STNK; STPA; SGND; SNDEQK; NDEQHK; NEQHRK; FVLIM; and HFY. These are all the positively scoring residue groups that occur in the Gonnet Pam250 matrix. The “strong” and “weak” residue groups are defined as “strong” score > 0.5 and “weak” score < 0.5, respectively.
[0082] FIGs. 2A-2E depict staining profiles of exemplary tested anti-TMPRSS2 antibodies AB1, AB2, AB4, and AB6 (FIGs. 2A-2D) compared to a commercially available antibody (FIG. 2E) on Caco-2 cells, assessed by flow cytometry.
[0083] FIGs. 3A-3F depict staining profiles of exemplary tested anti-TMPRSS2 antibodies
AB1, AB2, AB4, and AB6 (FIGs. 3B-3E) compared to a commercially available antibody (FIG. 3F) or control (FIG. 3 A) on HeLa cells, assessed by flow cytometry. [0084] FIGs. 4A-4C depict staining profiles of exemplary tested anti-TMPRSS2 antibodies AB1, AB2, AB4, and AB6 compared to control on lymphocytes (FIG. 4A), monocytes (FIG. 4B), or granulocytes (FIG. 4C), assessed by flow cytometry.
[0085] FIGs. 5A-5B depict staining of formalin-fixed paraffin-embedded (FFPE) samples with exemplary anti-human TMPRSS2 antibodies AB1, AB2, AB3, AB5 or AB7 on human kidney paraffin sections (FIG. 5 A) and non-TMPRSS2 expressing lymph node sections (FIG. 5B), assessed by immunohistochemistry.
[0086] FIGs. 6A-6D depict assessment of the functional activity of exemplary anti-human TMPRSS2 antibody AB1. The effect of AB1 on TMPRSS2 protease activity was compared to Nafamostat or control in both platelet-rich plasma (FIG. 6A) and using recombinant TMPRSS2 (FIG. 6B). The effect of AB1 on TMPRSS2 induced cell migration was compared to Nafamostat or control in CaCo-2 cells (FIG. 6C). The effect of AB1 on TMPRSS2 induced cell migration was compared to Nafamostat or four separate commercially available antibodies in CaCo-2 cells (FIG. 6D).
DESCRIPTION
[0087] Provided herein are antibodies that bind TMPRSS2, including antigen-binding fragments thereof, nucleic acids encoding such antibodies and antigen-binding fragments, and cells, such as recombinant cells for expressing and production of these antibodies and antigenbinding fragments that can bind to under physiological and/or in vitro conditions. Also provided are methods of producing and using the antibodies and antigen-binding fragments such as in methods for detecting TMPRSS2 in a sample from an individual, including methods for laboratory/ research purposes (e.g., flow cytometry, ELISA, and/or Western blot), and/or for the use and treatment and/or prevention of various diseases or disorders through the delivery of pharmaceutical or other compositions that contain such antibodies or antigen-binding fragments thereof.
[0088] All references cited herein, including patent applications, patent publications, and scientific literature and databases, are herein incorporated by reference in their entirety for all purposes to the same extent as if each individual reference were specifically and individually indicated to be incorporated by reference. [0089] For clarity of disclosure, and not by way of limitation, the detailed description is divided into the subsections that follow. The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.
I. Definitions
[0090] Unless defined otherwise, all terms of art, notations and other technical and scientific terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the subject matter pertains. 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 substantial difference over what is generally understood in the art. It is to be understood that the disclosure provided herein is not limited to particular compositions or biological systems. It is also understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not to be construed as limiting.
[0091] The term "antibody" as used herein includes antigen binding fragments thereof that retain binding specificity. For example, there are a number of well characterized antigen binding fragments. Thus, for example, pepsin digests an antibody C-terminal to the disulfide linkages in the hinge region to produce F(ab)'2, a dimer of Fab which itself is a light chain joined to VH-CH1 by a disulfide bond. The F(ab)'2 may be reduced under mild conditions to break the disulfide linkage in the hinge region thereby converting the (Fab')2 dimer into an Fab' monomer. The Fab' monomer is essentially an Fab with part of the hinge region (see, Fundamental Immunology, W.E. Paul, ed., Raven Press, N.Y. (1993), for a more detailed description of other antigen binding fragments). While various antigen binding fragments are defined in terms of the digestion of an intact antibody, one of skill will appreciate that fragments can be synthesized de novo either chemically or by utilizing recombinant DNA methodology. Thus, the term antibody, as used herein also includes antigen binding fragments either produced by the modification of whole antibodies or synthesized using recombinant DNA methodologies.
[0092] An antibody as described herein can consist of one or more polypeptides substantially encoded by immunoglobulin genes or fragments of immunoglobulin genes. The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA, IgD and IgE, respectively. In some embodiments, the antibody is IgG (e.g., IgGl, IgG2, IgG3, IgG4), IgM, IgA, IgD, or IgE.
[0093] A typical immunoglobulin (antibody) structural unit is known to comprise a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminus 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 chains respectively.
[0094] In an antibody, substitution variants have at least one amino acid residue removed and a different residue inserted in its place. The sites of greatest interest for substitutional mutagenesis include the hypervariable regions, but framework alterations are also contemplated. Examples of conservative substitutions are described above.
[0095] Substantial modifications in the biological properties of the antibody are accomplished by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a P-sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain. Naturally occurring residues are divided into groups based on common side-chain properties:
(1) Non-polar: Norleucine, Met, Ala, Vai, Leu, He;
(2) Polar without charge: Cys, Ser, Thr, Asn, Gin;
(3) Acidic (negatively charged): Asp, Glu;
(4) Basic (positively charged): Lys, Arg;
(5) Residues that influence chain orientation: Gly, Pro; and
(6) Aromatic: Trp, Tyr, Phe, His.
Non-conservative substitutions are made by exchanging a member of one of these classes for another class. [0096] One type of substitution that can be made is to change one or more cysteines in the antibody, which may be chemically reactive, to another residue, such as, without limitation, alanine or serine. For example, there can be a substitution of a non-canonical cysteine. The substitution can be made in a CDR or framework region of a variable domain or in the constant region of an antibody. In some embodiments, the cysteine is canonical (e.g., involved in disulfide bond formation). Any cysteine residue not involved in maintaining the proper conformation of the antibody also may be substituted, generally with serine, to improve the oxidative stability of the molecule and prevent aberrant cross-linking. Conversely, cysteine bond(s) may be added to the antibody to improve its stability, particularly where the antibody is an antigen binding fragment such as an Fv fragment.
[0097] Antibodies include VH-VL dimers, including single chain antibodies (antibodies that exist as a single polypeptide chain), such as single chain Fv antibodies (sFv or scFv) in which a variable heavy and a variable light domains are joined together (directly or through a peptide linker) to form a continuous polypeptide. The single chain Fv antibody is a covalently linked VH-VL which may be expressed from a nucleic acid including VH- and VL- encoding sequences either joined directly or joined by a peptide-encoding linker (e.g., Huston, et al. Proc. Nat. Acad. Sci. USA, 85:5879-5883, 1988). While the VH and VL are connected to each as a single polypeptide chain, the VH and VL domains associate non-covalently. Alternatively, the antibody can be another fragment. Other fragments can also be generated, e.g., using recombinant techniques, as soluble proteins or as fragments obtained from display methods. Antibodies can also include diantibodies and miniantibodies. Antibodies of the disclosure also include heavy chain dimers, such as antibodies from camelids. In some embodiments an antibody is dimeric. In other embodiments, the antibody may be in a monomeric form that has an active isotype. In some embodiments the antibody is in a multivalent form, e.g., a trivalent or tetravalent form.
[0098] An “antibody fragment” or “antigen binging fragment thereof’ comprises a portion of an intact antibody, the antigen binding and/or the variable region of the intact antibody. Atibody fragments or antigen binding fragments thereof, include but are not limited to Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, disulfide-linked Fvs (dsFv), Fd fragments, Fd' fragments; diabodies; linear antibodies (see U.S. Pat. No.5, 641, 870, Example 2; Zapata et al, Protein Eng. 8(10): 1057-1062 [1995]); single-chain antibody molecules, including single-chain Fvs (scFv) or single-chain Fabs (scFab); antigen-binding fragments of any of the above and multispecific antibodies from from antibody fragments.
[0099] "Fv" is composed of one heavy- and one light-chain variable region domain linked by non-covalent association. From the folding of these two domains emanate six complementarity determining regions (CDR) (3 in each from the heavy and light chain) that contribute the amino acid residues for antigen binding and confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although, in some cases, at a lower affinity than the entire binding site.
[0100] " dsFv" refers to an Fv with an engineered intermolecular disulfide bond, which stabilizes the VH-VL pair.
[0101] An "Fd fragment" is a fragment of an antibody containing a variable domain (VH) and one constant region domain (CHI) of an antibody heavy chain.
[0102] A "Fab fragment" is an antibody fragment that results from digestion of a full-length immunoglobulin with papain, or a fragment having the same structure that is produced synthetically, e.g., by recombinant methods. A Fab fragment contains a light chain (containing a VL and CL) and another chain containing a variable domain of a heavy chain (VH) and one constant region domain of the heavy chain (CHI).
[0103] A "F(ab')2 fragment" is an antibody fragment that results from digestion of an immunoglobulin with pepsin at pH 4.0-4.5, or a fragment having the same structure that is produced synthetically, e.g., by recombinant methods. The F(ab')2 fragment essentially contains two Fab fragments where each heavy chain portion contains an additional few amino acids, including cysteine residues that form disulfide linkages joining the two fragments.
[0104] A "Fab1 fragment" is a fragment containing one half (one heavy chain and one light chain) of the F(ab')2 fragment.
[0105] An "Fd1 fragment" is a fragment of an antibody containing one heavy chain portion of a F(ab')2 fragment. [0106] An "Fv1 fragment" is a fragment containing only the VH and VL domains of an antibody molecule.
[0107] An "scFv fragment" refers to an antibody fragment that contains a variable light chain (VL) and variable heavy chain (VH), covalently connected by a polypeptide linker in any order. The linker is of a length such that the two variable domains are bridged without substantial interference. Exemplary linkers are (Gly-Ser)n residues with some Glu or Lys residues dispersed throughout to increase solubility.
[0108] "Diabodies" are dimeric scFv; diabodies typically have shorter peptide linkers than scFvs, and preferentially dimerize.
[0109] As used herein, the terms “variable region” and “variable domain” refer to the portions of the light and heavy chains of an antibody that include amino acid sequences of complementary determining regions (CDRs, e.g., HCDR1, HCDR2, HCR3, LCDR1, LCDR2, and LCDR3) and framework regions (FRs). The variable domain for the heavy and light chains is commonly designated VH and VL, respectively. The variable domain is included on Fab, F(ab’)2, Fv and scFv antigen binding fragments described herein, and is involved in specific antigen recognition.
[0110] As used herein, "complementarity-determining region (CDR)" refers to the three hypervariable regions in each chain that interrupt the four framework regions established by the light and heavy chain variable regions. The CDRs are primarily responsible for binding to an epitope of an antigen. The CDRs of each chain are typically referred to as CDR1, CDR2, and CDR3, numbered sequentially starting from the N-terminus, and are also typically identified by the chain in which the particular CDR is located. Thus, a VH CDR3 is located in the variable domain of the heavy chain of the antibody in which it is found, whereas a VL CDR1 is the CDR1 from the variable domain of the light chain of the antibody in which it is found.
[OHl] The sequences of the framework regions of different light or heavy chains are relatively conserved within a species. The framework region of an antibody, that is the combined framework regions of the constituent light and heavy chains, serves to position and align the CDRs in three-dimensional space.
[0112] The amino acid sequences of the CDRs and framework regions can be determined using various well-known definitions in the art, e.g., Kabat, North method (see, e.g., North et al., J Mol Biol. 406(2):228-256, 2011), Chothia, international ImMunoGeneTics database (IMGT), and AbM (see, e.g., Johnson etal, supra, Chothia & Lesk, 1987, Canonical structures for the hypervariable regions of immunoglobulins. J. Mol. Biol. 196, 901-917; Chothia C. et al., 1989, Conformations of immunoglobulin hypervariable regions. Nature 342, 877-883; Chothia C. et al., 1992, structural repertoire of the human VH segments J. Mol. Biol. 227, 799- 817; Al-Lazikani et al., J.Mol.Biol 1997, 273(4)). Definitions of antigen combining sites are also described in the following: Ruiz et al., IMGT, the international ImMunoGeneTics database. Nucleic Acids Res., 28, 219-221 (2000); and Lefranc,M.-P. IMGT, the international ImMunoGeneTics database. Nucleic Acids Res. Jan l;29(l):207-9 (2001); MacCallum et al, Antibody-antigen interactions: Contact analysis and binding site topography, J. Mol. BioL, 262 (5), 732-745 (1996); and Martin et al, Proc. Natl Acad. Sci. USA, 86, 9268-9272 (1989); Martin, et al, Methods Enzymol., 203, 121-153, (1991); Pedersen et al, Immunomethods, 1, 126, (1992); and Rees et al, In Sternberg M.J.E. (ed.), Protein Structure Prediction. Oxford University Press, Oxford, 141-172 1996).
[0113] As used herein, "chimeric antibody" refers to an immunoglobulin molecule in which (a) the constant region, or a portion thereof, is altered, replaced or exchanged so that the antigen binding site (variable region) is linked to a constant region of a different or altered class, effector function and/or species, or an entirely different molecule which confers new properties to the chimeric antibody, e.g., an enzyme, toxin, hormone, growth factor, drug, etc.; or (b) the variable region, or a portion thereof, is altered, replaced or exchanged with a variable region, or portion thereof, having a different or altered antigen specificity; or with corresponding sequences from another species or from another antibody class or subclass.
[0114] As used herein, "humanized antibody" refers to an immunoglobulin molecule in CDRs from a donor antibody are grafted onto human framework sequences. Humanized antibodies may also comprise residues of donor origin in the framework sequences. The humanized antibody can also comprise at least a portion of a human immunoglobulin constant region. Humanized antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. Humanization can be performed using methods known in the art (e.g., Jones et al., Nature 321 :522-525; 1986; Riechmann et al., Nature 332:323-327, 1988; Verhoeyen etal., Science 239: 1534-1536, 1988); Presta, Curr. Op. Struct. Biol. 2:593-596, 1992; U.S. Patent No. 4,816,567), including techniques such as “superhumanizing" antibodies (Tan et al., J. Immunol. 169: 1119, 2002) and "resurfacing” (e.g., Staelens et al., Mol. Immunol. 43: 1243, 2006; and Roguska et al., Proc. Natl. Acad. Sci USA 91 : 969, 1994).
[0115] The term “recombinant” when used with reference, e.g., to a cell, or nucleic acid, protein, or vector, indicates that the cell, nucleic acid, protein or vector, has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found within the native (non-recombinant) form of the cell or express native genes that are otherwise abnormally expressed, under expressed or not expressed at all.
[0116] The terms “antigen,” “immunogen,” “antibody target,” “target analyte,” and like terms are used herein to refer to a molecule, compound, or complex that is recognized by an antibody, i.e., can be specifically bound by the antibody. The term can refer to any molecule that can be specifically recognized by an antibody, e.g., a polypeptide, polynucleotide, carbohydrate, lipid, chemical moiety, or combinations thereof (e.g., phosphorylated or glycosylated polypeptides, etc.). One of skill will understand that the term does not indicate that the molecule is immunogenic in every context, but simply indicates that it can be targeted by an antibody.
[0117] Antibodies bind to an “epitope” on an antigen. The epitope is the localized site on the antigen that is recognized and bound by the antibody. Epitopes can include a few amino acids or portions of a few amino acids, e.g., 5 or 6, or more, e.g., 20 or more amino acids, or portions of those amino acids. In some cases, the epitope includes non-protein components, e.g., from a carbohydrate, nucleic acid, or lipid. In some cases, the epitope is a three- dimensional moiety. Thus, for example, where the target is a protein, the epitope can be comprised of consecutive amino acids, or amino acids from different parts of the protein that are brought into proximity by protein folding (e.g., a discontinuous epitope). The same is true for other types of target molecules that form three-dimensional structures. An epitope typically includes at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial conformation. Methods of determining spatial conformation of epitopes include, for example, x-ray crystallography and 2-dimensional nuclear magnetic resonance. See, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, Glenn E. Morris, Ed (1996). [0118] A “label” or a “detectable moiety” is a diagnostic agent or component detectable by spectroscopic, radiological, photochemical, biochemical, immunochemical, chemical, or other physical means. Exemplary labels include radiolabels (e.g., inIn, "mTc, 131I, 67Ga) and other FDA-approved imaging agents. Additional labels include 32P, fluorescent dyes, electron-dense reagents, enzymes, biotin, digoxigenin, or haptens and proteins or other entities which can be made detectable, e.g., by incorporating a radiolabel into the targeting agent. Any method known in the art for conjugating a nucleic acid or nanocarrier to the label may be employed, e.g., using methods described in Hermanson, Bioconjugate Techniques 1996, Academic Press, Inc., San Diego.
[0119] A “labeled” or “tagged” antibody or agent is one that is bound, either covalently, through a linker or a chemical bond, or noncovalently, through ionic, van der Waals, electrostatic, or hydrogen bonds to a label such that the presence of the antibody or agent may be detected by detecting the presence of the label bound to the antibody or agent.
[0120] Techniques for conjugating detectable and therapeutic agents to antibodies are well known (see, e.g., Amon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., "Antibodies For Drug Delivery"in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review" in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); and Thorpe et al., "The Preparation And Cytotoxic Properties Of Antibody- Toxin Conjugates", Immunol. Rev., 62: 119-58 (1982)).
[0121] The terms “specific for,” “specifically binds,” and like terms refer to a molecule (e.g., antibody or antigen binding fragment) that binds to a target with at least 2-fold greater affinity than non-target compounds, e.g., at least any of 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 25-fold, 50-fold, or 100-fold greater affinity. For example, an antibody that specifically binds a target (e.g., TMPRSS2) will typically bind the target with at least a 2-fold greater affinity than a non-target. Specificity can be determined using standard methods, e.g., solid-phase ELISA immunoassays (see, e.g., Harlow & Lane, Using Antibodies, A Laboratory Manual (1998) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity). [0122] The term “binds” with respect to an antibody target (e.g., antigen, analyte, immune complex), typically indicates that an antibody binds a majority of the antibody targets in a pure population (assuming appropriate molar ratios). For example, an antibody that binds a given antibody target typically binds to at least 2/3 of the antibody targets in a solution (e.g., at least any of 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100%). One of skill will recognize that some variability will arise depending on the method and/or threshold of determining binding.
[0123] A “control” sample or value refers to a sample that serves as a reference, usually a known reference, for comparison to a test sample. For example, a test sample can be taken from a test condition, e.g., in the presence of a test compound, and compared to samples from known conditions, e.g., in the absence of the test compound (negative control), or in the presence of a known compound (positive control). A control can also represent an average value or a range gathered from a number of tests or results. One of skill in the art will recognize that controls can be designed for assessment of any number of parameters. For example, a control can be devised to compare therapeutic benefit based on pharmacological data (e.g., half-life) or therapeutic measures (e.g., comparison of benefit and/or side effects). Controls can be designed for in vitro applications. One of skill in the art will understand which controls are valuable in a given situation and be able to analyze data based on comparisons to control values. Controls are also valuable for determining the significance of data. For example, if values for a given parameter are widely variant in controls, variation in test samples will not be considered as significant.
[0124] The terms “identical” or percent “identity,” in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region, when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual alignment and visual inspection (see, e.g., NCBI web site ncbi.nlm.nih.gov/BLAST/ or the like). Such sequences are then said to be “substantially identical.” As described below, the preferred algorithms can account for gaps and the like. Preferably, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or more preferably over a region that is 50-100 or more amino acids or nucleotides in length.
[0125] 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 entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Preferably, default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.
[0126] A “comparison window”, as used herein, includes reference to a segment of any one of the number of contiguous positions selected from the group consisting of from 20 to 600, usually about 50 to about 200, more usually about 100 to about 150 in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are well- known in the art.
[0127] An algorithm that is suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. 25:3389-3402 (1977) and Altschul et al., J. Mol. Biol. 215:403-410 (1990), respectively. BLAST and BLAST 2.0 are used, with the parameters described herein, to determine percent sequence identity for the nucleic acids and proteins of the disclosure. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov/). This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always > 0) and N (penalty score for mismatching residues; always < 0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negativescoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M=5, N=-4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89: 10915 (1989)) alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands.
[0128] The term “nucleic acid” refers to deoxyribonucleotides or ribonucleotides and polymers thereof in either single- or double-stranded form, and complements thereof. The term encompasses nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).
[0129] Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions) and complementary sequences, as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer c/ al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); Rossolini etal., Mol. Cell. Probes 8:91-98 (1994)).
[0130] The terms “polypeptide,” “peptide” and “protein” are used interchangeably herein to refer to a polymer of amino acid residues. The terms encompass to amino acid polymers in which one or more amino acid residue is an artificial chemical mimetic of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non- naturally occurring amino acid polymer. [0131] The term “amino acid” refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified, e.g., hydroxyproline, y- carboxyglutamate, and O-phosphoserine. Amino acid analogs refers to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Amino acid mimetics refers to chemical compounds that have a structure that is different from the general chemical structure of an amino acid, but that functions in a manner similar to a naturally occurring amino acid.
[0132] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes.
[0133] The term “compete”, as used herein with regard to an antibody, means that a first antibody, or an antigen-binding portion thereof, competes for binding with a second antibody, or an antigen-binding portion thereof, where binding of the first antibody with its cognate epitope is detectably decreased in the presence of the second antibody compared to the binding of the first antibody in the absence of the second antibody. The alternative, where the binding of the second antibody to its epitope is also detectably decreased in the presence of the first antibody, can, but need not be the case. That is, a first antibody can inhibit the binding of a second antibody to its epitope without that second antibody inhibiting the binding of the first antibody to its respective epitope. However, where each antibody detectably inhibits the binding of the other antibody with its cognate epitope or ligand, whether to the same, greater, or lesser extent, the antibodies are said to “cross-compete” with each other for binding of their respective epitope(s). Both competing and cross-competing antibodies are encompassed by the present disclosure. Regardless of the mechanism by which such competition or crosscompetition occurs (e.g., steric hindrance, conformational change, or binding to a common epitope, or portion thereof, and the like), the skilled artisan would appreciate, based upon the teachings provided herein, that such competing and/or cross-competing antibodies are encompassed and can be useful for the methods disclosed herein.
[0134] Numerous types of competitive binding assays are known, for example: solid phase direct or indirect radioimmunoassay (RIA), solid phase direct or indirect enzyme immunoassay (EIA), sandwich competition assay (see Stahli et al., Methods in Enzymology 9:242-253 (1983)); solid phase direct biotin-avidin EIA (see Kirkland et al., J. Immunol. 137:3614-3619 (1986)); solid phase direct labeled assay, solid phase direct labeled sandwich assay (see Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Press (1988)); solid phase direct label RIA using 1-125 label (see Morel et al., Molec. Immunol. 25(1):7-15 (1988)); solid phase direct biotin-avidin EIA (Cheung et al., Virology 176:546-552 (1990)); and direct labeled RIA (Moldenhauer et al., Scand. J. Immunol. 32:77-82 (1990)). Typically, such an assay involves the use of purified antigen bound to a solid surface or cells bearing either of these, an un-labelled test immunoglobulin and a labeled reference immunoglobulin. Competitive inhibition is measured by determining the amount of label bound to the solid surface or cells in the presence of the test immunoglobulin. Usually the test immunoglobulin is present in excess. Antibodies identified by competition assay (competing antibodies) 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. Usually, when a competing antibody is present in excess, it will inhibit specific binding of a reference antibody to a common antigen by at least 50 or 75%.
[0135] The term “TMPRSS2” as used herein refers to human TMPRSS2 proteins, isoforms or variants thereof, including naturally occurring variants of human TMPRSS2, such as splice variants or allelic variants. The amino acid sequence of an exemplary human TMPRSS2 is shown in SEQ ID NO: 1. The amino acid sequence of another exemplary human TMPRSS2 is shown in SEQ ID NO: 2. In some embodiments, human TMPRSS2 can refer to a variant, such as an allelic variant or splice variant, that exhibits at least or at least about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to any of SEQ ID NOs: 1-2. In some embodiments, it is understood that the provided antibodies or antigen-binding fragments may exhibit cross-reactive binding to another mammalian TMPRSS2 protein, such as murine TMPRSS2, or a primate TMPRSS2.
Human TMPRSS2 isoform 1 amino acid sequence (SEQ ID NO:1; UniProt 015393-1) MALNSGSPPAIGPYYENHGYQPENPYPAQPTVVPTVYEVHPAQYYPSPVPQYAPRVL TQASNPVVCTQPKSPSGTVCTSKTKKALCITLTLGTFLVGAALAAGLLWKFMGSKCS NSGIECDSSGTCINPSNWCDGVSHCPGGEDENRCVRLYGPNFILQVYSSQRKSWHPV CQDDWNENYGRAACRDMGYKNNFYSSQGIVDDSGSTSFMKLNTSAGNVDIYKKLY HSDACSSKAVVSLRCIACGVNLNSSRQSRIVGGESALPGAWPWQVSLHVQNVHVCG GSIITPEWIVTAAHCVEKPLNNPWHWTAFAGILRQSFMFYGAGYQVEKVISHPNYDS KTKNNDIALMKLQKPLTFNDLVKPVCLPNPGMMLQPEQLCWISGWGATEEKGKTSE VLNAAKVLLIETQRCNSRYVYDNLITPAMICAGFLQGNVDSCQGDSGGPLVTSKNNI WWLIGDTSWGSGCAKAYRPGVYGNVMVFTDWIYRQMRADG
Human TMPRSS2 isoform 2 amino acid sequence (SEQ ID NO:2; UniProt 015393-2)
MPPAPPGGESGCEERGAAGHIEHSRYLSLLDAVDNSKMALNSGSPPAIGPYYENHGY QPENPYPAQPTVVPTVYEVHPAQYYPSPVPQYAPRVLTQASNPVVCTQPKSPSGTVC TSKTKKALCITLTLGTFLVGAALAAGLLWKFMGSKCSNSGIECDSSGTCINPSNWCD GVSHCPGGEDENRCVRLYGPNFILQVYSSQRKSWHPVCQDDWNENYGRAACRDMG YKNNFYSSQGIVDDSGSTSFMKLNTSAGNVDIYKKLYHSDACSSKAVVSLRCIACGV NLNSSRQSRIVGGESALPGAWPWQVSLHVQNVHVCGGSIITPEWIVTAAHCVEKPLN NPWHWTAFAGILRQSFMFYGAGYQVEKVISHPNYDSKTKNNDIALMKLQKPLTFND LVKPVCLPNPGMMLQPEQLCWISGWGATEEKGKTSEVLNAAKVLLIETQRCNSRYV YDNLITPAMICAGFLQGNVDSCQGDSGGPLVTSKNNIWWLIGDTSWGSGCAKAYRP GVYGNVMVFTDWIYRQMRADG
[0136] In some embodiments, any of the exemplary human TMPRSS2 amino acid sequences comprises a cytoplasmic domain. In some embodiments, the cytoplasmic domain comprises the sequence set forth in the SEQ ID NO: 3. In some embodiments, any of the exemplary human TMPRSS2 amino acid sequences comprises a transmembrane domain. In some embodiments, the transmembrane domain comprises the sequence set forth in the SEQ ID NO: 4. In some embodiments, any of the exemplary human TMPRSS2 amino acid sequences comprises an extracellular domain. In some embodiments, the extracellular domain comprises the sequence set forth in the SEQ ID NO: 5.
[0137] By "solid support" is meant a non-aqueous matrix to which an antibody according to the provided disclosure can adhere or attach. For example, solid supports include, but are not limited to, a microtiter plate, a membrane (e.g. , nitrocellulose), a bead, a dipstick, a thin-layer chromatographic plate, or other solid medium.
[0138] As used herein, an "individual" or a "subject" is a mammal. A "mammal" for purposes of treatment includes humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, horses, rabbits, cattle, pigs, hamsters, gerbils, mice, ferrets, rats, cats, etc. In some embodiments, the individual or subject is human.
IL Antibodies that bind TMPRSS2
[0139] Provided herein are antibodies, including antigen binding fragments thereof, that specifically bind to TMPRSS2. In some embodiments, the provided antibodies include monoclonal antibodies and antigen-binding fragments thereof that bind TMPRSS2 that provide superior target specificity, signal-to-noise ratios, and the like as compared to other reported antibodies. Also provided herein are methods for producing anti-TMPRSS2 antibodies, and methods for detecting and using such antibodies.
[0140] Transmembrane serine protease 2 (TMPRSS2) is s type II transmembrane protein containing approximately 492 amino acids encoded by the TMPRSS2 gene. The TMPRSS2 gene is widely conserved and has two isoforms, which differ only in the N-terminal, cytoplasmic tail, with isoform 1 containing 37 additional amino acids on the tail which are not present in isoform 2. Both TMPRSS2 isoforms are autocatalytically activated from the inactive zymogen form.
[0141] TMPRSS2 has been associated with physiological and pathological processes such as digestion, tissue remodeling, blood coagulation, fertility, inflammatory responses, tumor cell invasion, apoptosis and pain, and is upregulated by androgenic hormones in prostate cancer cells. TMPRSS2 activates several substrates that include pro-hepatocyte growth factor/HGF, the protease activated receptor-2/F2RLl or matriptase/ST14 leading to extracellular matrix disruption and metastasis of prostate cancer cells. TMPRSS2 has been suggested to facilitate human coronaviruse SARS-CoV and SARS-CoV-2 infections via two independent mechanisms: proteolytic cleavage of ACE2 receptor which promotes viral uptake, and cleavage of coronavirus spike glycoproteins which activates the glycoprotein for host cell entry. In addition, TMPRSS2 has been shown to activate the spike glycoproteins of human coronavirus 229E (HCoV-229E) and human coronavirus EMC (HCoV-EMC) and the fusion glycoproteins F0 of Sendai virus (SeV), human metapneumovirus (HMPV), human parainfluenza 1, 2, 3, 4a and 4b viruses (HPIV). Both SARS-CoV and SARS-CoV-2 (COVID-19) use angiotensinconverting enzyme 2 (ACE2) and TMPRSS2 to facilitate entry to cells, but with SARS-CoV-2 human-to-human transmission is much higher than SARS-CoV (Thunders et al. J Clin Pathol 73(12):773-776 (2020); Lucas et al. Cancer Discovery 4:1310-1325 (2014); Wilson et al. Biochem J. 388-967-972 (2005); Ko et al Cancer Res. 75:2949-2960 (2015)).
[0142] In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of TMPRSS2. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the extracellular domain of TMPRSS2. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the extracellular domain of TMPRSS2 set forth in SEQ ID NO: 5. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the transmembrane domain of TMPRSS2. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the transmembrane domain of TMPRSS2 set forth in SEQ ID NO: 4. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the cytoplasmic domain of TMPRSS2. In some embodiments, any of the antibodies of antigen binding fragments thereof provided herein bind all or a portion of the cytoplasmic domain of TMPRSS2 set forth in SEQ ID NO: 3.
[0143] In some embodiments, any of the antibodies or antigen binding fragments thereof is a TMPRSS2 antibody or antigen binding fragment thereof. In some embodiments, the antibody or antigen binding fragment thereof is isolated (e.g., separated from a component of its natural environment (e.g., an animal, a biological sample)). In some embodiments, the anti- antibody is a humanized antibody, or an antigen binding fragment thereof. In some embodiments, the antibody is a derivative of a humanized antibody that binds. In some embodiments, the antibody binds under laboratory conditions (e.g., binds in vitro, binds in a flow cytometry assay, binds in an ELISA). In some embodiments, the antibody binds under physiological conditions (e.g., binds in a cell in a subject).
[0144] Generally, the antibodies provided herein comprise at least one immunoglobulin heavy chain variable domain and at least one immunoglobulin light chain variable domain. In some embodiments, an antibody described herein comprises two immunoglobulin heavy chain variable domains and two immunoglobulin light chain variable domains. Typically, each immunoglobulin heavy chain variable domain of the antibody comprises first, second, and third heavy chain complementarity determining regions (CDRs; HCDR1, HCDR2, and HCDR3), and each immunoglobulin light chain variable domain of the antibody comprises first, second, and third light chain CDRs (LCDR1, LCDR2, and LCDR3).
[0145] In some embodiments, the antibodies are antigen binding fragments such as Fab, F(ab’)2, Fv or scFv. The antigen binding fragments can be generated using any means known in the art including, chemical digestion (e.g., papain or pepsin) and recombinant methods. Methods for isolating and preparing recombinant nucleic acids are known to those skilled in the art (see, Sambrook et al., Molecular Cloning. A Laboratory Manual (2d ed. 1989); Ausubel et al., Current Protocols in Molecular Biology (1995)). The antibodies can be expressed in a variety of host cells, including E. coli, other bacterial hosts, yeast, and various higher eukaryotic cells such as the COS, CHO, and HeLa cells lines and myeloma cell lines.
[0146] The disclosure provides an isolated antibody or antigen binding fragment thereof that specifically binds to CD56, wherein the isolated antibody comprises: a) an immunoglobulin heavy chain variable domain comprising: (i) a heavy chain complementary determining region 1 (CDRH1) comprising the sequence comprising the sequence X1X2X3X4X5 (SEQ ID NO: 81), wherein Xi is S, N or D; X2 is Y or S; X3 is G, W or D; X4 is V, M or I; and X5 is S, Q or N; (ii) a heavy chain complementary determining region 2 (CDRH2) comprising the sequence X1IX2X3X4X5X6X7X8X9X10X11X12X13X14X15X16 (SEQ ID NO: 82), wherein Xi is V, A, Y or W; X2 is W,Y,S or N; X3 is G, P, S or T; X4 is D, V, G or Y; X5 is G, D or T; X6 is S or G; X7 is T, D, R, S or E; Xs is N, T, I or P; X9 is Y, R, K or T; X10 is H, Y or F; Xu is S, T, P, N or A; X12 is A, Q, D or E; X13 is L, K, T or G; Xi4 is I, F or V; X15 is S or K; and Xi6 is G or no amino acid; and (iii) a heavy chain complementary determining region 3 (CDRH3) comprising the sequence X1X2X3X4X5X6X7X8X9X10X11X12X13 (SEQ ID NO: 83), wherein Xi is P, L, D, A , S or I; X2 is G, S, F, Y or H; X3 is N, P, D, Y or G; X4 is N, S, Y, R, D or G; X5 is Y, N F or no amino acid; Xe is D, Y S or no amino acid; X7 is W, D, Y, H or no amino acid; Xs is Y, F, G, A, W or no amino acid; X9 is F, D, A, M, Y or no amino acid; X10 is D, C or no amino acid; Xu is V, F, D or no amino acid; X12 is D, V or no amino acid; and X13 is Y or no amino acid; and b) an immunoglobulin light chain variable domain comprising: (i) a light chain complementary determining region 1 (CDRL1) comprising the sequence X1ASX2X3IX4X5X6X7X8 (SEQ ID NO: 84) wherein Xi is K or R; X2 is Q or E; X3 is D, S, E or N; X4 is N, G, S or Y; X5 is K, T, S or V; Xe is Y or N; X7 is I, M or L; and Xs is A, H, S or T; (ii) a light chain complementary determining region 2 (CDRL2) comprising the sequence X1X2X3X4X5X6X7 (SEQ ID NO: 85) wherein Xi is Y, R or A; X2 is T, A or G; X3 is S, N or T; X4 is T, E, R or N; X5 is L or S; Xe is Q, I, E, D or A; and X7 is P, S or D; and (iii) a light chain complementary determining region 3 (CDRL3) comprising the sequence X1X2X3X4X5X6X7X8X9 (SEQ ID NO: 86), wherein Xi is L or Q; X2 is Q or H; X3 is Y, S or F; X4 is A, Y, D, H or W; X5 is N, E, S or G; X6 is L, W, P, Y or T; X7 is L or P; Xs is T, L or Y; and X9 is T or no amino acid.
[0147] In some embodiments, antibodies of the disclosure can comprise sequences of a heavy chain complementary determining region 1 (HCDR1), an HCDR2, an HCDR3, a light chain complementary determining region 1 (LCDR1), a LCDR2, a LCDR3. Exemplary CDR amino acid sequences and associated SEQ ID NOs are set forth in Table 1, and exemplary amino acid sequences for heavy chain variable domains (VH), and light chain variable domains( VL) and associated SEQ ID NOs are set forth in Table 2.
Table 1. Amino acid sequences of CDRs from anti-TMPRSS2 antibodies shown according to Kabat numbering.
Figure imgf000050_0001
Figure imgf000051_0001
Table 2. Amino acid sequences of Variable Heavy (VHs) and variable light (VL) domains from anti-TMPRSS2 antibodies.
Figure imgf000051_0002
Atorney Docket: 102738-1363851-002410PC
[0148] In some embodiments, any of the antibody or antigen binding fragments thereof of the disclosure comprises: (1) an CDRH1 having a sequence of any one of SEQ ID NOS:45, 46, 47, 48, 49 and 50 or a variant thereof that has a sequence having one amino acid substitution relative to a sequence of any one of SEQ ID NOS: 45, 46, 47, 48, 49 and 50; (2) an CDRH2 having a sequence of any one of SEQ ID NOS: 51, 52, 53, 54, 55 and 56 a variant thereof that has a sequence having one, two, or three amino acid substitutions relative to a sequence of any one of SEQ ID NOS: 51, 52, 53, 54, 55 and 56; (3) an CDRH3 having a sequence of any one of SEQ ID NOS: 57, 58, 59, 60, 61, 62 and 63, or a variant thereof that has a sequence having one or two amino acid substitutions relative to a sequence of any one of SEQ ID NOS: 57, 58, 59, 60, 61, 62 and 63; (4) a CDRL1 having a sequence of any one of SEQ ID NOS: 64, 65, 66, 67, 68 and 69 a variant thereof that has a sequence having one or two amino acid substitutions relative to a sequence of any one of SEQ ID NOS: 64, 65, 66, 67, 68 and 69; (5) a CDRL2 having a sequence of any one of SEQ ID NOS:70, 71, 72, 73, and 74 or a variant thereof that has a sequence having one substitution relative to a sequence of any one of SEQ ID NOS: 70, 71, 72, 73, and 74; and (6) a CDRL3 having a sequence of any one of SEQ ID NOS: 75, 76, 77, 78, 79 and 80, or a variant thereof that has a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 75, 76, 77, 78, 79 and 80.
[0149] In some embodiments, the immunoglobulin heavy chain variable domain comprises a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 51, 52, 53, 54, 55 or 56; and a CDRH3 comprising the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63.
[0150] In some embodiments, the immunoglobulin heavy chain variable domain comprises: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56; and a CDRH3 comprising the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63.
[0151] In some embodiments, the immunoglobulin light chain variable domain comprises: a CDRL1 comprising the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 70, 71, 72, 73 or 74; and a CDRL3 comprising the sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 75, 76, 77, 78, 79 or 80.
[0152] In some embodiments, the immunoglobulin light chain variable domain comprises: a CDRL1 comprising a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74; and a CDRL3 comprising a sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
[0153] In some embodiments, the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein contains an immunoglobulin heavy chain variable domain with the amino acid sequence set forth in any of SEQ ID NOS:20, 21, 22, 23, 24, 25 or 26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20, 21, 22, 23, 24, 25 or 26. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 20, 21, 22, 23, 24, 25 or 26.
[0154] In some embodiments, the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein contains an immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS:27, 28, 29, 30, 31, 32 or 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27, 28, 29, 30, 31, 32 or 33. In some of any embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 27, 28, 29, 30, 31, 32 or 33.
[0155] In some of any embodiments, any of the antibodies or antigen binding fragments thereof further comprises a signal peptide. In some embodiments, the immunoglobulin heavy chain variable domain further comprises a signal peptide of any of SEQ ID NOS: 34-38. In some of any embodiments, the immunoglobulin light chain variable domain further comprises a signal peptide of SEQ ID NO: 39-44.
AB1
[0156] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:45 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:45; (2) a CDRH2 having the sequence of SEQ ID NO:51 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:51; (3) a CDRH3 having the sequence of SEQ ID NO:57 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:57. In some embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:64 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:64; (5) a CDRL2 having the sequence of SEQ ID NO:70 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:75 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 75.
[0157] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45; (2) a CDRH2 having the sequence of SEQ ID NO:51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; (3) a CDRH3 having the sequence of SEQ ID NO:57 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:57. In some embodiments, the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64; (5) a CDRL2 having the sequence of SEQ ID NO:70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:75 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 75.
[0158] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:45; (2) an CDRH2 having the sequence of SEQ ID NO: 51; (3) an CDRH3 having the sequence of SEQ ID NO: 57; (4) a CDRL1 having the sequence of SEQ ID NO: 64; (5) a CDRL2 having the sequence of SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:75.
[0159] In some embodiments, the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:20 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:20. In some embodiments, the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:20. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:27.
[0160] In certain embodiments, the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:20 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:20; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:27 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:27.
[0161] In some embodiments, the antibody or antigen binding fragment thereof can comprise
(1) a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:45, 51 and 57, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 45, 51 and 57 and
(2) a light chain variable domain having a CDRL1, a CDRL2, and a CDRL3 of SEQ ID NOS:64, 70 and 75, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 64, 70 and 75. Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
AB2
[0162] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:46 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:46; (2) a CDRH2 having the sequence of SEQ ID NO: 52 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:52; (3) a CDRH3 having the sequence of SEQ ID NO:58 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 58. In some embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:65 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 65; (5) a CDRL2 having the sequence of SEQ ID NO:71 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:71; and (6) a CDRL3 having the sequence of SEQ ID NO:76 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:76.
In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:46 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46; (2) a CDRH2 having the sequence of SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:52; (3) a CDRH3 having the sequence of SEQ ID NO:58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 58. In some embodiments, the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:65 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 65; (5) a CDRL2 having the sequence of SEQ ID NO:71 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:71; and (6) a CDRL3 having the sequence of SEQ ID NO:76 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 76.
In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:46; (2) an CDRH2 having the sequence of SEQ ID NO:52; (3) an CDRH3 having the sequence of SEQ ID NO:58; (4) a CDRL1 having the sequence of SEQ ID NO: 65; (5) a CDRL2 having the sequence of SEQ ID NO:71; and (6) a CDRL3 having the sequence of SEQ ID NO:76.
In some embodiments, the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:21 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:21. In some embodiments, the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:21. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:28. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:28.
[0163] In certain embodiments, the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:21 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:21; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:28 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:28.
[0164] In some embodiments, the antibody or antigen binding fragment thereof can comprise
(1) a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:46, 52 and 58, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 46, 52 and 58 and
(2) a light chain variable domain having a CDRL1, a CDRL2, and a CDRL3 of SEQ ID NOS:65, 71 and 76, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 65, 71 and 76. Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
AB3
[0165] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:47 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:47; (2) a CDRH2 having the sequence of SEQ ID NO:53 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:53; (3) a CDRH3 having the sequence of SEQ ID NO:58 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 58. In some embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:66 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 66; (5) a CDRL2 having the sequence of SEQ ID NO: 72 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:72; and (6) a CDRL3 having the sequence of SEQ ID NO:77 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:77. [0166] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:47 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47; (2) a CDRH2 having the sequence of SEQ ID NO:53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53; (3) a CDRH3 having the sequence of SEQ ID NO:59 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:59. In some embodiments, the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:66 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:66; (5) a CDRL2 having the sequence of SEQ ID NO:72 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:72; and (6) a CDRL3 having the sequence of SEQ ID NO:77 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:77.
[0167] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:47; (2) an CDRH2 having the sequence of SEQ ID NO:53; (3) an CDRH3 having the sequence of SEQ ID NO:59; (4) a CDRL1 having the sequence of SEQ ID NO: 66; (5) a CDRL2 having the sequence of SEQ ID NO:72; and (6) a CDRL3 having the sequence of SEQ ID NO:77.
[0168] In some embodiments, the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:22 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:22. In some embodiments, the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:22. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:29. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:29.
[0169] In certain embodiments, the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:22 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:22; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:29 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:29.
[0170] In some embodiments, the antibody or antigen binding fragment thereof can comprise (1) a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:47, 53 and 59, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 47, 53 and 59and (2) a light chain variable domain having a CDRL1, a CDRL2, and a CDRL3 of SEQ ID NOS: 66, 72 and 77, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 66, 72 and 77. Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
AB4
[0171] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:48 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:48; (2) a CDRH2 having the sequence of SEQ ID NO: 54 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:54; (3) a CDRH3 having the sequence of SEQ ID NO:60 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:60. In some embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:67 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 67; (5) a CDRL2 having the sequence of SEQ ID NO: 73 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:78.
[0172] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:48 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48; (2) a CDRH2 having the sequence of SEQ ID NO:54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:54; (3) a CDRH3 having the sequence of SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:60. In some embodiments, the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:67 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:67; (5) a CDRL2 having the sequence of SEQ ID NO:73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:78.
[0173] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:48; (2) an CDRH2 having the sequence of SEQ ID NO: 54; (3) an CDRH3 having the sequence of SEQ ID NO: 60; (4) a CDRL1 having the sequence of SEQ ID NO: 67; (5) a CDRL2 having the sequence of SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78.
[0174] In some embodiments, the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:23 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:23. In some embodiments, the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:23. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:30. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:30.
[0175] In certain embodiments, the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:20 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:20; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:27 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:27.
[0176] In some embodiments, the antibody or antigen binding fragment thereof can comprise
(1) a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:48, 54 and 60, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 48, 54 and 60 and
(2) a light chain variable domain having a CDRL1, a CDRL2, and a CDRL3 of SEQ ID NOS:67, 73 and 78, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 67, 73 and 78. Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
AB5
[0177] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:45 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:45; (2) a CDRH2 having the sequence of SEQ ID NO:51 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:51; (3) a CDRH3 having the sequence of SEQ ID NO:61 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:61. In some embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:64 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:64; (5) a CDRL2 having the sequence of SEQ ID NO:70 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:79 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:79.
[0178] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45; (2) a CDRH2 having the sequence of SEQ ID NO:51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; (3) a CDRH3 having the sequence of SEQ ID NO:61 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:61. In some embodiments, the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64; (5) a CDRL2 having the sequence of SEQ ID NO:70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:79 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:79.
[0179] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:45; (2) an CDRH2 having the sequence of SEQ ID NO:51; (3) an CDRH3 having the sequence of SEQ ID NO:61; (4) a CDRL1 having the sequence of SEQ ID NO: 64; (5) a CDRL2 having the sequence of SEQ ID NO:70; and (6) a CDRL3 having the sequence of SEQ ID NO:79. In some embodiments, the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:24 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:24. In some embodiments, the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:24. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:31. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:31.
[0180] In certain embodiments, the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:24 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:24; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:31 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 31.
[0181] In some embodiments, the antibody or antigen binding fragment thereof can comprise
(1) a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:45, 51 and 61, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 45, 51 and 61 and
(2) a light chain variable domain having a CDRL1, a CDRL2, and a CDRL3 of SEQ ID NOS:64, 70 and 79, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 64, 70 and 79. Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
AB6
[0182] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:49 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:49; (2) a CDRH2 having the sequence of SEQ ID NO: 55 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:55; (3) a CDRH3 having the sequence of SEQ ID NO:62 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:62. In some embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:68 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:68; (5) a CDRL2 having the sequence of SEQ ID NO:73 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:78.
In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:49 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49; (2) a CDRH2 having the sequence of SEQ ID NO:55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:55; (3) a CDRH3 having the sequence of SEQ ID NO:62 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:62. In some embodiments, the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:68 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:68; (5) a CDRL2 having the sequence of SEQ ID NO:73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 78.
In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:49; (2) an CDRH2 having the sequence of SEQ ID NO:55; (3) an CDRH3 having the sequence of SEQ ID NO:62; (4) a CDRL1 having the sequence of SEQ ID NO:68; (5) a CDRL2 having the sequence of SEQ ID NO:73; and (6) a CDRL3 having the sequence of SEQ ID NO:78.
In some embodiments, the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:25 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:25. In some embodiments, the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:25. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:32, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:32. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:32.
[0183] In certain embodiments, the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:35 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:25; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:32 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:32.
[0184] In some embodiments, the antibody or antigen binding fragment thereof can comprise
(1) a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:49, 55 and 62, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 49, 55 and 62; and
(2) a light chain variable domain having a CDRL1, a CDRL2, and a CDRL3 of SEQ ID NOS:68, 73 and 78, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 68, 73 and 78. Such an antibody can be an IgGl, IgG2, IgG3, or IgG4. AB7
[0185] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:50 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:50; (2) a CDRH2 having the sequence of SEQ ID NO:56 or a sequence having one, two or three amino acid substitutions relative to the sequence of SEQ ID NO:56; (3) a CDRH3 having the sequence of SEQ ID NO:63 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:63. In some embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:69 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO: 69; (5) a CDRL2 having the sequence of SEQ ID NO: 74 or a sequence having one amino acid substitution relative to the sequence of SEQ ID NO:74; and (6) a CDRL3 having the sequence of SEQ ID NO:80 or a sequence having one or two amino acid substitutions relative to the sequence of SEQ ID NO:80.
[0186] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) a CDRH1 having the sequence of SEQ ID NO:50 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:50; (2) a CDRH2 having the sequence of SEQ ID NO:56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56; (3) a CDRH3 having the sequence of SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 63. In some embodiments, the antibody or antigen binding fragment thereof of the disclosure can comprise: (4) a CDRL1 having the sequence of SEQ ID NO:69 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:69; (5) a CDRL2 having the sequence of SEQ ID NO:74 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:74; and (6) a CDRL3 having the sequence of SEQ ID NO:80 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:80.
[0187] In particular embodiments, an antibody or antigen binding fragment thereof of the disclosure can comprise: (1) an CDRH1 having the sequence of SEQ ID NO:50; (2) an CDRH2 having the sequence of SEQ ID NO:56; (3) an CDRH3 having the sequence of SEQ ID NO:63; (4) a CDRL1 having the sequence of SEQ ID NO: 69; (5) a CDRL2 having the sequence of SEQ ID NO:74; and (6) a CDRL3 having the sequence of SEQ ID NO:80.
[0188] In some embodiments, the antibody or antigen binding fragment thereof of any of the embodiments disclosed herein can include an immunoglobulin heavy chain variable domain comprising the amino acid sequence set forth in SEQ ID NO:26 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:26. In some embodiments, the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:26. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:33. In some embodiments, the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:33.
[0189] In certain embodiments, the antibody or antigen binding fragment thereof comprises (i) a heavy chain comprising a heavy chain variable domain having the sequence of SEQ ID NO:26 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:26; and (ii) a light chain comprising a light chain variable domain having the sequence of SEQ ID NO:33 and a sequence having at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO:33.
[0190] In some embodiments, the antibody or antigen binding fragment thereof can comprise
(1) a heavy chain variable domain having an CDRH1, an CDRH2, and an CDRH3 of SEQ ID NOS:50, 56 and 63, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 50, 56 and 63; and
(2) a light chain variable domain having a CDRL1, a CDRL2, and a CDRL3 of SEQ ID NOS:69, 74 and 80, respectively, and at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 69, 74 and 80. Such an antibody can be an IgGl, IgG2, IgG3, or IgG4.
IV. Fc polypeptide
[0191] An antibody provided herein can comprise a fragment crystallizable region (Fc region), also referred to as an Fc polypeptide herein. An Fc polypeptide is part of each of the two heavy chains in the antibody and can interact with certain cell surface receptors and certain components of the complement system. An Fc polypeptide typically includes the CH2 domain and the CH3 domain, which are immunoglobulin constant region domain polypeptides. In some embodiments, the Fc polypeptide in an antibody described herein can be a wild-type Fc polypeptide, e.g., a human IgGl Fc polypeptide. In certain embodiments, an antibody described herein can comprise a wild-type Fc polypeptide having the sequence of SEQ ID NO:87:
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
[0192] In other embodiments, an antibody described herein can comprise a variant of the wild-type Fc polypeptide that has at least 90% (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%) identity to the sequence of a wild-type Fc polypeptide (e.g., SEQ ID NO:87) and at least one amino acid substitution relative to the sequence of a wild-type Fc polypeptide (e.g., SEQ ID NO:87).
[0193] In some embodiments, an Fc polypeptide includes one or more modifications (e.g., one or more amino acid substitutions, insertions, or deletions relative to a comparable wildtype Fc region). Antibodies comprising modified Fc polypeptides typically have altered phenotypes relative to antibodies comprising wild-type Fc polypeptides. For example, antibodies comprising modified Fc polypeptides can have altered serum half-life, altered stability, altered susceptibility to cellular enzymes, and/or altered effector function (e.g., as assayed in an NK-dependent or macrophage-dependent assay). [0194] In some embodiments, an Fc polypeptide in an antibody described herein can include amino acid substitutions that modulate effector function. In certain embodiments, an Fc polypeptide in an antibody described herein can include amino acid substitutions that reduce or eliminate effector function. Illustrative Fc polypeptide amino acid substitutions that reduce effector function include, but are not limited to, substitutions in a CH2 domain, e.g., at positions 4 and 5 (position numbering relative to the sequence of SEQ ID NO:47) (see, e.g., Lund et al., J Immunol. 147(8):2657-62, 1991). For example, in some embodiments, one or both Fc polypeptides in an antibody described herein can comprise L4A and L5A substitutions.
[0195] Additional Fc polypeptide amino acid substitutions that modulate an effector function include, e.g., substitution at position 99 (position numbering relative to the sequence of SEQ ID NO:5). For example, in some embodiments, one or both Fc polypeptides in an antibody described herein can comprise P99G substitution. In certain embodiments, one or both Fc polypeptides in an antibody described herein can have L4A, L5A, and P99G substitutions.
[0196] In some embodiments, an Fc polypeptide includes one or more modifications that alter (relative to a wild-type Fc polypeptide) the Ratio of Affinities of the modified Fc polypeptide to an activating FcyR (such as FcyRIIA or FcyRIIIA) relative to an inhibiting FcyR (such as FcyRIIB):
Wild-Type to Variant Change in Affinity to FcyR A
Ratio of Affinities = .
Figure imgf000070_0001
Wild-Type to Variant Change in Affinity to FcyR
Figure imgf000070_0002
[0197] Where a modified Fc polypeptide has a Ratio of Affinities greater than 1, an antibody herein may have particular use in providing a therapeutic or prophylactic treatment of a disease, disorder, or infection, or the amelioration of a symptom thereof, where an enhanced efficacy of effector cell function (e.g., ADCC) mediated by FcyR is desired, e.g., cancer or infectious disease. Where a modified Fc region has a Ratio of Affinities less than 1, an antibody herein may have particular use in providing a therapeutic or prophylactic treatment of a disease or disorder, or the amelioration of a symptom thereof, where a decreased efficacy of effector cell function mediated by FcyR is desired, e.g., autoimmune or inflammatory disorders. Table 2 lists examples of single, double, triple, quadruple, and quintuple amino acid substitutions in an Fc polypeptide that provide a Ratio of Affinities greater than 1 or less than 1 (see e.g., PCT Publication Nos. WO 04/063351; WO 06/088494; WO 07/024249; WO 06/113665; WO 7/021841; WO 07/106707; WO 2008/140603). Amino acid positions are numbered according o EU numbering scheme.
Table 2
Figure imgf000071_0001
Figure imgf000072_0001
III. Antibodies that competitively bind with an anti-TMPRSS2 antibody
[0198] Also provided herein are antibodies that competitively bind, or are capable of competitively binding (e.g., competitor antibodies), with one or more TMPRSS2 antibodies described herein. In certain instances, an antibody (i.e., competitor antibody) may be considered to compete for binding to when the competitor binds to the same general region of as an antibody described herein. In certain instances, an antibody (i.e., competitor antibody) may be considered to compete for binding to when the competitor binds to the exact same region of as an antibody described herein (e.g., exact same peptide (linear epitope) or exact same surface amino acids (conformational epitope)). In certain instances, an antibody (i.e., competitor antibody) may be considered capable of competing for binding to when the competitor binds to the same general region of as an antibody described herein (i.e., extracellular region or leucine-rich binding domain) under suitable assay conditions. In certain instances, an antibody (i.e., competitor agent) may be considered capable of competing for binding to when the competitor binds to the exact same region of as an antibody described herein (e.g., exact same peptide (linear epitope) or exact same surface amino acids (conformational epitope)) under suitable assay conditions.
[0199] In certain instances, an antibody (i.e., competitor antibody) may be considered to compete for binding to TMPRSS2 when the competitor blocks the binding of one or more antibodies described herein to TMPRSS2, for example, under suitable assay conditions. Whether a competitor blocks the binding of one or more antibodies described herein to may be determined using a suitable competition assay or blocking assay, such as, for example, a blocking assay as described in herein. A competitor antibody may block binding of one or more antibodies described herein to in a competition or blocking assay by 50% or more (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, or 100%), and conversely, one or more antibodies described herein may block binding of the competitor antibody to in a competition or blocking assay by about 50% or more (e.g., e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more, or 100%).
[0200] In certain instances, an antibody (i.e., competitor antibody) may be considered to compete for binding to TMPRSS2 when the competitor binds to with a similar affinity as one or more antibodies described herein, for example, under suitable assay conditions. In some embodiments, an antibody (i.e., competitor antibody) is considered to compete for binding to when the competitor binds to with an affinity that is at least about 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%) of the affinity of one or more antibodies described herein.
[0201] Also provided herein are antibodies that bind to, or are capable of binding to, the same epitope as one or more antibodies described herein. In particular, provided herein are antibodies that compete with one or more antibodies described herein for binding to the same epitope (e.g., same peptide (linear epitope) or same surface amino acids (conformational epitope)) on TMPRSS2. Such antibodies that bind the same epitope may be referred to as epitope competitors.
IV. Polyclonal and monoclonal antibodies
[0202] Polyclonal antibodies may be raised in animals (vertebrate or invertebrates, including mammals, birds and fish, including cartilaginous fish) by multiple subcutaneous (sc) or intraperitoneal (ip) injections of a relevant antigen and an adjuvant. It may be useful to conjugate the relevant antigen to a protein or other carrier that is immunogenic in the species to be immunized, e.g., keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, or soybean trypsin inhibitor using a bifunctional or derivatizing agent, for example, maleimidobenzoyl sulfosuccinimide ester (conjugation through cysteine residues), N- hydroxysuccinimide (through lysine residues), glutaraldehyde, succinic anhydride, SOC12, or R1N=C=NR, where R and R1 are different alkyl groups. Non-protein carriers (e.g., colloidal gold) also may be used for antibody production.
[0203] Animals can be immunized against the antigen, immunogenic conjugates, or derivatives by combining, e.g., 100 pg or 5 pg of the protein or conjugate (for rabbits or mice, respectively) with three volumes of Freund's complete adjuvant and injecting the solution intradermally at multiple sites. One month later the animals are boosted with one-fifth to one- tenth of the original amount of peptide or conjugate in Freund's complete adjuvant by subcutaneous injection at multiple sites. Seven to 14 days later the animals are bled and the serum is assayed for antibody titer. Animals are boosted until the titer plateaus. Often, the animal is boosted with the conjugate of the same antigen, but conjugated to a different protein and/or through a different cross-linking reagent. Conjugates also can be made in recombinant cell culture as protein fusions. Also, aggregating agents such as alum are suitably used to enhance the immune response.
[0204] Monoclonal antibodies may be made using a hybridoma, e.g., the hybridoma method first described by Kohler et al., Nature, 256:495 (1975), or may be made by other methods such as recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567). In the hybridoma method, a mouse or other appropriate host animal, such as a hamster or macaque monkey, is immunized to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the protein used for immunization. Alternatively, lymphocytes may be immunized in vitro. Lymphocytes then are fused with myeloma cells using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell (see, e.g., Goding, Monoclonal Antibodies: Principles and Practice, pp.59-103 (Academic Press, 1986)).
[0205] The hybridoma cells thus prepared are seeded and grown in a suitable culture medium that may contain one or more substances that inhibit the growth or survival of the unfused, parental myeloma cells. For example, if the parental myeloma cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (HAT medium), which substances prevent the growth of HGPRT-deficient cells. Preferred myeloma cells are those that fuse efficiently, support stable high-level production of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. Among these, preferred myeloma cell lines are murine myeloma lines, such as SP-2 or X63- Ag8-653 cells available from the American Type Culture Collection, Rockville, Md. USA. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol., 133:3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp. 51-63 (Marcel Dekker, Inc., New York, 1987)).
[0206] Culture medium in which hybridoma cells are growing is assayed for production of monoclonal antibodies directed against the antigen. The binding specificity of monoclonal antibodies produced by hybridoma cells may be determined by immunoprecipitation, by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbant assay (ELISA), or by flow cytometric analysis of cells expressing the membrane antigen. The binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson et al., Anal. Biochem., 107:220 (1980). [0207] After hybridoma cells are identified that produce antibodies of the desired specificity, affinity, and/or activity, the clones may be subcloned by limiting dilution procedures and grown by standard methods (see, e.g., Goding, Monoclonal Antibodies: Principles and Practice, pp.59-103 (Academic Press, 1986)). Suitable culture media for this purpose include, for example, D-MEM or RPMI-1640 medium. In addition, the hybridoma cells may be grown in vivo as ascites tumors in an animal. The monoclonal antibodies secreted by the subclones are suitably separated from the culture medium, ascites fluid, or serum by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
[0208] DNA encoding the monoclonal antibodies can 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 monoclonal antibodies). Alternatively, cDNA may be prepared from mRNA and the cDNA then subjected to DNA sequencing. The hybridoma cells serve as a preferred source of such genomic DNA or RNA for cDNA preparation. Once isolated, the DNA may be placed into expression vectors, which are well known in the art, and which are then transfected into host cells such as E. coll cells, simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells.
V. Humanization and amino acid variants
[0209] General methods for humanization of antibodies are described, for example, in U.S. Patent Nos. 5861155, 6479284, 6407213, 6639055, 6500931, 5530101, 5585089, 5693761, 5693762, 6180370, 5714350, 6350861, 5777085, 5834597, 5882644, 5932448, 6013256, 6129914, 6210671, 6329511, 5225539, 6548640, and 5624821. In certain embodiments, it may be desirable to generate amino acid sequence variants of these humanized antibodies, particularly where these improve the binding affinity or other biological properties (e.g., halflife) of the antibody.
[0210] In some embodiments, the antibody is a humanized antibody, i.e., an antibody that retains the reactivity of a non-human antibody while being less immunogenic in humans. This can be achieved, for instance, by retaining the non-human CDR regions and replacing the remaining parts of the antibody with their human counterparts. See, e.g., Morrison et al., PNAS USA, 81 :6851-6855 (1984); Morrison and Oi, Adv. Immunol., 44:65-92 (1988); Verhoeyen et al., Science, 239: 1534-1536 (1988); Padlan, Molec. Immun., 28:489-498 (1991); Padlan, Molec. Immun., 31(3): 169-217 (1994). Techniques for humanizing antibodies are well known in the art and are described in e.g., U.S. Patent Nos. 4,816,567; 5,530,101; 5,859,205; 5,585,089; 5,693,761; 5,693,762; 5,777,085; 6,180,370; 6,210,671; and 6,329,511; WO 87/02671; EP Patent Application 0173494; Jones et al. (1986) Nature 321 :522; and Verhoyen et al. (1988) Science 239: 1534. Humanized antibodies are further described in, e.g., Winter and Milstein ( \ 99 \ ) Nature 349:293. For example, polynucleotides comprising a first sequence coding for humanized immunoglobulin framework regions and a second sequence set coding for the desired immunoglobulin complementarity determining regions can be produced synthetically or by combining appropriate cDNA and genomic DNA segments. Human constant region DNA sequences can be isolated in accordance with well-known procedures from a variety of human cells. The CDRs for producing the immunoglobulins of the present disclosure can be similarly derived from monoclonal antibodies capable of specifically binding to .
[0211] Amino acid sequence variants of the antibody can be prepared by introducing appropriate nucleotide changes into the antibody DNA, or by peptide synthesis. Such variants include, for example, deletions from, and/or insertions into and/or substitutions of, residues within the amino acid sequences of the antibodies for the examples herein. Any combination of deletion, insertion, and substitution is made to arrive at the final construct, provided that the final construct possesses the desired characteristics. The amino acid changes also may alter post-translational processes of the humanized or variant antibody, such as changing the number or position of glycosylation sites.
[0212] One method for identification of certain residues or regions of the antibody that are preferred locations for mutagenesis is called “alanine scanning mutagenesis,” as described by, e.g., Cunningham and Wells, Science, 244: 1081-1085 (1989). Here, a residue or group of target residues are identified (e.g., charged residues such as Arg, Asp, His, Lys, and Glu) and replaced by a neutral or negatively charged amino acid (most preferably Ala or poly-Ala) to affect the interaction of the amino acids with antigen. Those amino acid locations demonstrating functional sensitivity to the substitutions then are refined by introducing further or other variants at, or for, the sites of substitution. Thus, while the site for introducing an amino acid sequence variation is predetermined, the nature of the mutation per se need not be predetermined. For example, to analyze the performance of a mutation at a given site, alanine scanning or random mutagenesis is conducted at the target codon or region and the expressed antibody variants are screened for the desired activity. Amino acid sequence insertions include amino- and/or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include an N-terminal methionyl residue or the antibody fused to an epitope tag. Other insertional variants include the fusion of an enzyme or a polypeptide that increases the serum half-life of the antibody to the N- or C- terminus of the antibody.
[0213] Another type of variant is an amino acid substitution variant. These variants have at least one amino acid residue removed from the antibody molecule and a different residue inserted in its place. The sites of greatest interest for substitutional mutagenesis include the hypervariable regions, but FR alterations are also contemplated. Conservative substitutions are preferred, but more substantial changes may be introduced and the products may be screened. Examples of substitutions are listed below:
Ala (A): Vai; Leu; He; Vai
Arg (R): Lys; Gin; Asn; Lys
Asn (N): Gin; His; Asp, Lys; Gin; Arg
Asp (D): Glu; Asn
Cys (C): Ser; Ala
Gin (Q): Asn; Glu
Glu (E): Asp; Gin
Gly (G): Ala
His (H): Asn; Gin; Lys; Arg
He (I): Leu; Vai; Met; Ala; Leu; Phe; Norleucine
Leu (L): Norleucine; He; Vai; lie; Met; Ala; Phe Lys (K): Arg; Gin; Asn
Met (M): Leu; Phe; He
Phe (F): Leu; Vai; He; Ala; Tyr
Pro (P): Ala
Ser (S): Thr
Thr (T): Ser
Trp (W): Tyr; Phe
Tyr (Y): Trp; Phe; Thr; Ser
Vai (V): He; Leu; Met; Phe; Ala; Norleucine
[0214] Substantial modifications in the biological properties of an antibody are accomplished by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain. Naturally occurring residues are divided into groups based on common sidechain properties:
(1) hydrophobic: Norleucine, Met, Ala, Vai, Leu, He;
(2) neutral hydrophilic: Cys, Ser, Thr;
(3) acidic: Asp, Glu;
(4) basic: Asn, Gin, His, Lys, Arg;
(5) residues that influence chain orientation: Gly, Pro; and
(6) aromatic: Trp, Tyr, Phe
[0215] Non-conservative substitutions will entail exchanging a member of one of the above classes for another class. [0216] Any cysteine residue not involved in maintaining the proper conformation of the antibody also may be substituted, to improve the oxidative stability of the molecule and prevent aberrant crosslinking. Conversely, cysteine bond(s) may be added to the antibody to improve its stability (particularly where the antibody is an antigen binding fragment such as an Fv fragment).
[0217] One type of substitutional variant involves substituting one or more hypervariable region residues of a parent antibody. Generally, the resulting variant(s) selected for further development will have improved biological properties relative to the parent antibody from which they are generated. A convenient way for generating such substitutional variants is affinity maturation using phage display. Briefly, several hypervariable region sites (e.g., 6-7 sites) are mutated to generate all possible amino acid substitutions at each site. The antibody variants thus generated can be displayed in the monovalent fashion from filamentous phage particles as fusions to the gene III product of M13 packaged within each particle. The phage- displayed variants are then screened for their biological activity (e.g., binding affinity) as herein disclosed. In order to identify candidate hypervariable region sites for modification, alanine- scanning mutagenesis can be performed to identify hypervariable region residues contributing significantly to antigen binding. Alternatively, or in addition, it may be beneficial to analyze a crystal structure of the antigen-antibody complex to identify contact points between the antibody and antigen. Such contact residues and neighboring residues are candidates for substitution according to the techniques elaborated herein. Once such variants are generated, the panel of variants is subjected to screening as described herein and antibodies with superior properties in one or more relevant assays may be selected for further development.
[0218] Another type of amino acid variant of the antibody alters the original glycosylation pattern of the antibody. By altering is meant deleting one of more carbohydrate moieties found in the antibody, and/or adding one or more glycosylation sites that are not present in the antibody. Glycosylation of antibodies is typically either N-linked and/or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the most common recognition sequences for enzymatic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O- linked glycosylation refers to the attachment of one of the sugars N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5- hydroxyproline or 5-hydroxylysine may also be used. Addition of glycosylation sites to the antibody can be accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). The alteration may also be made by the addition of, or substitution by, one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites).
VI. Other modifications
[0219] Other modifications of an antibody are contemplated. For example, technology herein also pertains to immunoconjugates comprising an antibody described herein conjugated to a cytotoxic agent such as a toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant or animal origin, or fragments thereof), or a radioactive isotope (for example, a radioconjugate), or a cytotoxic drug. Such conjugates are sometimes referred to as “antibodydrug conjugates” or “ADC.” Conjugates can be made using a variety of bifunctional protein coupling agents such asN-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bis-azido compounds (such as bis-(p-azidobenzoyl)hexanediamine), bis-diazonium derivatives (such as bis-(p- diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as tolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as l,5-difluoro-2,4-dinitrobenzene).
[0220] In some of any embodiments, any of the antibodies or antigen binding fragments thereof disclosed herein may be formulated as immunoliposomes. Liposomes containing an antibody are prepared by methods know in the art, such as described in Epstein et al., Proc. Natl. Acad. Sci. USA 82:3688 (1985); Hwang et al., Proc. Natl. Acad. Sci. USA 77:4030 (1980); and U.S. Pat. Nos. 4,485,045 and 4,544,545. Liposomes with enhanced circulation time are disclosed in U.S. Pat. No. 5,013,556. For example, liposomes can be generated by the reverse phase evaporation method with a lipid composition comprising phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to yield liposomes with the desired diameter. Fab' fragments of an antibody provided herein can be conjugated to the liposomes as described in Martin et al., J. Biol. Chem. 257:286-288 (1982) via a disulfide interchange reaction. Another active ingredient is optionally contained within the liposome. [0221] Enzymes or other polypeptides can be covalently bound to an antibody by techniques well known in the art such as the use of the heterobifunctional cross-linking reagents discussed above. In some embodiments, fusion proteins comprising at least the antigen binding region of an antibody provided herein linked to at least a functionally active portion of an enzyme can be constructed using recombinant DNA techniques well known in the art (see, e.g., Neuberger et al., Nature 312:604-608 (1984)).
[0222] In certain embodiments, it may be desirable to use an antigen binding fragment, rather than an intact antibody, to increase penetration of target tissues and cells, for example. In such instances, it may be desirable to modify the antigen binding fragment in order to increase its serum half-life. This may be achieved, for example, by incorporation of a salvage receptor binding epitope into the antigen binding fragment (e.g., by mutation of the appropriate region in the antigen binding fragment or by incorporating the epitope into a peptide tag that is then fused to the antigen binding fragment at either end or in the middle, e.g., by DNA or peptide synthesis; see, e.g., WO96/32478 published Oct. 17, 1996).
[0223] In some embodiments, any of the antibodies or antigen fragments thereof disclosed herein are conjugated or hybridized to an oligonucleotide. In some embodiments, the oligonucleotide includes a sample barcode sequence, a binding site for a primer and an anchor. In some embodiments, the oligonucleotide can be conjugated or hybridized to any of the detectable markers or labels disclosed herein. . In some embodiments, the oligonucleotide is a polymeric sequence. In some embodiments, the terms “oligonucleotide” and “polynucleotide” are used interchangeably to refer to a single-stranded multimer of nucleotides from about 2 to about 500 nucleotides in length. In some embodiments, any of the oligonucleotides described herein can be synthetic, made enzymatically (e.g., via polymerization), or using a “split-pool” method. In some embodiments, any of the oligonucleotides described herein can include ribonucleotide monomers (i.e., can be oligoribonucleotides) and/or deoxyribonucleotide monomers (i.e., oligodeoxyribonucleotides). In some embodiments, any of the oligonucleotides described herein can include a combination of both deoxyribonucleotide monomers and ribonucleotide monomers in the oligonucleotide (e.g., random or ordered combination of deoxyribonucleotide monomers and ribonucleotide monomers). In some embodiments, the oligonucleotide can be 4 to 10, 10 to 20, 21 to 30, 31 to 40, 41 to 50, 51 to 60, 61 to 70, 71 to 80, 80 to 100, 100 to 150, 150 to 200, 200 to 250, 250 to 300, 300 to 350, 350 to 400, or 400-500 nucleotides in length. In some embodiments, any of the oligonucleotides described herein can include one or more functional moieties that are attached (e.g., covalently or non-covalently) to another structure. In some embodiments, any of the oligonucleotides described herein can include one or more detectable labels (e.g., a radioisotope or fluorophore). In some embodiments, the anchor is a defined polymer, e.g., a polynucleotide or oligonucleotide sequence, which is designed to hybridize to a complementary oligonucleotide sequence. In some embodimentsthe anchor is designed for the purpose of generating a double stranded construct oligonucleotide sequence. In some embodiments, the anchor is positioned at the 3’ end of the construct oligonucleotide sequence. In other embodiments, the anchor is positioned at the 5’ end of the construct oligonucleotide sequence. Each anchor is specific for its intended complementary sequence.
[0224] In some embodiments, the sample barcode sequence is a polymer, e.g., a polynucleotide, which when it is a functional element, is specific for a single ligand. In some embodiments, the sample barcode sequence can be used for identifying a particular cell or substrate, e.g., Drop-seq microbead. In some embodiments, the sample barcode sequence can be formed of a defined sequence of DNA, RNA, modified bases or combinations of these bases, as well as any other polymer defined above. In some embodiments, the sample barcode sequence is about 2 to 4 monomeric components, e.g., nucleotide bases, in length. In other embodiments, the barcode is at least about 1 to 100 monomeric components, e.g., nucleotides, in length. Thus in various embodiments, the barcode is formed of a sequence of at least 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, 50, 51,
52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76,
77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 80, 91, 92, 93, 94, 95, 96, 97, 98, 99 or up to
100 monomeric components, e.g., nucleic acids. In some embodiments, the sample barcode sequence is a particular barcode that can be unique relative to other barcodes.
[0225] In some of any embodiments, the sample barcode sequences can have a variety of different formats. For example, sample barcode sequences can include polynucleotide barcodes, random nucleic acid and/or amino acid sequences, and synthetic nucleic acid and/or amino acid sequences. A sample barcode sequence can be attached to an analyte or to another moiety or structure in a reversible or irreversible manner. A sample barcode sequences can be added to, for example, a fragment of a deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) sample before or during sequencing of the sample. Sample barcode sequences can allow for identification and/or quantification of individual sequencing-reads (e.g., a barcode can be or can include a unique molecular identifier or “UMI”).
[0226] Sample barcode sequences can spatially-resolve molecular components found in biological samples, for example, at single-cell resolution (e.g., a barcode can be or can include a “spatial barcode”). In some embodiments, a barcode includes both a UMI and a spatial barcode. In some embodiments, a barcode includes two or more sub-barcodes that together function as a single barcode. For example, a polynucleotide barcode can include two or more polynucleotide sequences (e.g., sub-barcodes) that are separated by one or more non-barcode sequences.
[0227] In some embodiments, the binding site for a primer is a functional component of the oligonucleotide which itself is an oligonucleotide or polynucleotide sequence that provides an annealing site for amplification of the oligonucleotide. The binding site for a primer can be formed of polymers of DNA, RNA, PNA, modified bases or combinations of these bases, or polyamides, etc. In some embodiments, the binding site for a primer is about 10 of such monomeric components, e.g., nucleotide bases, in length. In other embodiments, the binding site for a primer is at least about 5 to 100 monomeric components, e.g., nucleotides, in length. Thus in various embodiments, the binding site for a primer is formed of a sequence of at least 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, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 80, 91, 92, 93, 94, 95, 96, 97, 98, 99 or up to 100 monomeric components, e.g., nucleic acids. In certain embodiments, the binding site for a primer can be a generic sequence suitable as a annealing site for a variety of amplification technologies. Amplification technologies include, but are not limited to, DNA-polymerase based amplification systems, such as polymerase chain reaction (PCR), real-time PCR, loop mediated isothermal amplification (LAMP, MALBAC), strand displacement amplification (SDA), multiple displacement amplification (MDA), recombinase polymerase amplification (RPA) and polymerization by any number of DNA polymerases (for example, T4 DNA polymerase, Sulfulobus DNA polymerase, Klenow DNA polymerase, Bst polymerase, Phi29 polymerase) and RNA-polymerase based amplification systems (such as T7-, T3-, and SP6-RNA- polymerase amplification), nucleic acid sequence based amplification (NASBA), selfsustained sequence replication (3 SR), rolling circle amplification (RCA), ligase chain reaction (LCR), helicase dependent amplification (I), ramification amplification method and RNA-seq. Methods for conjugating or hybridizing an oligonucleotide can be performed in a manner set forth in WO/2018/144813, WO/2016/018960, WO/2018/089438, WO/2014/182528, WO/2018/026873, WO/2021/188838.
[0228] In some embodiments, a modification can optionally be introduced into the antibodies (e.g., within the polypeptide chain or at either the N- or C-terminal), e.g., to extend in vivo half-life, such as PEGylation or incorporation of long-chain polyethylene glycol polymers (PEG). Introduction of PEG or long chain polymers of PEG increases the effective molecular weight of the polypeptides, for example, to prevent rapid filtration into the urine. In some embodiments, a lysine residue in the sequence is conjugated to PEG directly or through a linker. Such linker can be, for example, a Glu residue or an acyl residue containing a thiol functional group for linkage to the appropriately modified PEG chain. An alternative method for introducing a PEG chain is to first introduce a Cys residue at the C-terminus or at solvent exposed residues such as replacements for Arg or Lys residues. This Cys residue is then site- specifically attached to a PEG chain containing, for example, a maleimide function. Methods for incorporating PEG or long chain polymers of PEG are known in the art (described, for example, in Veronese, F. M., et al., Drug Disc. Today 10: 1451-8 (2005); Greenwald, R. B., et al., Adv. Drug Deliv. Rev. 55: 217-50 (2003); Roberts, M. J., et al., Adv. Drug Deliv. Rev., 54: 459-76 (2002)), the contents of which are incorporated herein by reference.
[0229] Covalent modifications of an antibody are also included within the scope of this technology. For example, modifications may be made by chemical synthesis or by enzymatic or chemical cleavage of an antibody. Other types of covalent modifications of an antibody are introduced into the molecule by reacting targeted amino acid residues of the antibody with an organic derivatizing agent that is capable of reacting with selected side chains or the N- or C- terminal residues. Example covalent modifications of polypeptides are described in U.S. Pat. No. 5,534,615, specifically incorporated herein by reference. A preferred type of covalent modification of the antibody comprises linking the antibody to one of a variety of non- proteinaceous polymers, e.g., polyethylene glycol, polypropylene glycol, or polyoxyalkylenes, in the manner set forth in, e.g., U.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192 or 4,179,337. VII. Nucleic acids, vectors, host cells, and recombinant methods
[0230] The disclosure also provides isolated nucleic acids encoding an antibody, vectors and host cells comprising the nucleic acid, and recombinant techniques for the production of the antibody. A nucleic acid herein may include one or more subsequences, each referred to as a polynucleotide.
[0231] Provided herein are nucleic acids (e.g., isolated nucleic acids) comprising a nucleotide sequence that encodes an antibody, or fragment thereof. In some embodiments, a nucleic acid encodes an immunoglobulin heavy chain variable domain of an antibody provided herein. In some embodiments, a nucleic acid encodes an immunoglobulin light chain variable domain of an antibody provided herein. In some embodiments, a nucleic acid encodes an immunoglobulin heavy chain variable domain and an immunoglobulin light chain variable domain of an antibody provided herein. In some embodiments, a nucleic acid comprises a nucleotide sequence that encodes an amino acid sequence of any one of SEQ ID NOS: 6-12 or 13-19.
[0232] In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in any of SEQ ID NOs: 6-12. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 6. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 7. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 8. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 9. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 10. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 11. In some of any embodiments, the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 12.
[0233] In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in any of SEQ ID NOs: 13-19. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 13. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 14. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 15. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 16. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 17. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 18. In some of any embodiments, the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 19.
[0234] Provided herein is a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain and the immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any of any of the antibodies or antigen binding fragments provided herein.
[0235] In some of any embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 6; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 13. In some of any embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 7; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 14. In some of any embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 8; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 15. In some of any embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 9; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 16. In some of any embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 10; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 17. In some of any embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 11; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 18. In some of any embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 12; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 19.
[0236] In some of any embodiments, the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in any of SEQ ID NOs:6-12; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in any of SEQ ID NOs: 13-19.
[0237] For recombinant production of an antibody, a nucleic acid encoding the antibody may be isolated and inserted into a replicable vector for further cloning (amplification of the DNA) or for expression. In certain instances, an antibody may be produced by homologous recombination. DNA encoding an antibody can 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). Many vectors are available. The vector components generally include, but are not limited to, one or more of the following: a signal sequence, and origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence.
[0238] Suitable host cells for cloning or expressing DNA in vectors herein can be prokaryote, yeast, or higher eukaryote cells. Suitable prokaryotes for this purpose include eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as Escherichia, e.g., E. coli, Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium, Serratia, e.g., Serratia marcescans, and Shigella, as well as Bacilli such as //. subtilis and B. licheniformis, Pseudomonas such as /< aeruginosa, and Streptomyces. One preferred E. coli cloning host is E. coli 294 (ATCC 31,446), although other strains such as E. coli B, E. coli X1776 (ATCC 31,537), and E. coli W3110 (ATCC 27,325) can also be suitable. These examples are illustrative rather than limiting.
[0239] In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for antibody-encoding vectors. Saccharomyces cerevisiae, or common baker’s yeast, is the most commonly used among lower eukaryotic host microorganisms. A number of other genera, species, and strains are commonly available and useful herein, such as Schizosaccharomyces pom be , Kluyveromyces hosts such as, e.g., K. lactis, K. fragilis (ATCC 12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K. waltii (ATCC 56,500), K. drosophilarum (ATCC 36,906), K. thermotolerans, and K. marxianus: yarrowia (EP 402,226); Pichia pastoris (EP 183,070); Candida,' Trichoderma reesia (EP 244,234); Neurospora crassa: Schwanniomyces such as Schwanniomyces occidenlahs and filamentous fungi such as, e.g., Neurospora, Penicillium, Tolypocladium, and Aspergillus hosts such as A. nidulans and A. niger.
[0240] Suitable host cells for the expression of antibodies can also be derived from multicellular organisms. Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains and variants and corresponding permissive insect host cells from hosts such as Spodoptera frugiperda (caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito), Drosophila melanogaster (fruitfly), and Bombyx mori (silk moth) have been identified. A variety of viral strains for transfection are publicly available, e.g., the L-l variant of Autographa californica NPV and the Bm-5 strain of Bombyx mori NPV, and such viruses may be used as the virus herein according to the present technology, particularly for transfection of Spodoptera frugiperda cells. Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato, and tobacco can also be utilized as hosts.
[0241] Suitable host cells for the expression of antibodies also may include vertebrate cells (e.g., mammalian cells). Vertebrate cells may be propagated in culture (tissue culture). Examples of useful mammalian host cell lines include monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BEK, ATCC CCL 10); Chinese hamster ovary cells/-DHFR (CHO, Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); mouse Sertoli cells (TM4, Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL 70); African green monkey kidney cells (VERO-76, ATCC CRL-1587); human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3 A, ATCC CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells (Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51); TRI cells (Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982)); MRC 5 cells; FS4 cells; and a human hepatoma line (Hep G2). [0242] Host cells may be transformed with the above-described expression or cloning vectors for antibody production and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences. Host cells used to produce antibodies provided herein may be cultured in a variety of media. Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPMI-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing the host cells. In addition, any of the media described in Ham et al., Meth. Enz. 58:44 (1979), Barnes et al., Anal. Biochem.102:255 (1980), U.S. Pat. Nos. 4,767,704; 4,657,866; 4,927,762; 4,560,655; or 5,122,469; WO 90/03430; WO 87/00195; or U.S. Pat. Re. 30,985 may be used as culture media for the host cells. Any of these media may be supplemented as necessary with hormones and/or other growth factors (such as insulin, transferrin, or epidermal growth factor), salts (such as sodium chloride, calcium, magnesium, and phosphate), buffers (such as HEPES), nucleotides (such as adenosine and thymidine), antibiotics (such as GENTAMYCIN™), trace elements (defined as inorganic compounds usually present at final concentrations in the micromolar range), and glucose or an equivalent energy source. Any other necessary supplements may also be included at appropriate concentrations that would be known to those skilled in the art. The culture conditions, such as temperature, pH, and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.
[0243] When using recombinant techniques, antibodies can be produced intracellularly, in the periplasmic space, or directly secreted into the medium. If the antibody is produced intracellularly, as a first step, the particulate debris, either host cells or lysed fragments, is removed, for example, by centrifugation or ultrafiltration. Carter et al., Bio/Technology 10: 163-167 (1992) describe a procedure for isolating antibodies that are secreted to the periplasmic space of E. coli. Briefly, cell paste is thawed in the presence of sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min. Cell debris can be removed by centrifugation. Where the antibody is secreted into the medium, supernatants from such expression systems are generally first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit. A protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants. [0244] The antibody composition prepared from the cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography, with affinity chromatography being the preferred purification technique. The suitability of protein A as an affinity ligand depends on the species and isotype of any immunoglobulin Fc domain that is present in the antibody. Protein A can be used to purify antibodies that are based on human heavy chains (Lindmark et al., J. Immunol. Meth. 62: 1-13 (1983)). Protein G is recommended for all mouse isotypes and for human y3 (Guss et al., EMBO J. 5: 15671575 (1986)). The matrix to which the affinity ligand is attached is most often agarose, but other matrices are available. Mechanically stable matrices such as controlled pore glass or poly(styrenedivinyl)benzene allow for faster flow rates and shorter processing times than can be achieved with agarose. Where the antibody comprises a CH3 domain, Bakerbond ABX.TM. resin (J. T. Baker, Phillipsburg, N.J.) is useful for purification. Other techniques for protein purification, such as fractionation on an ion-exchange column, ethanol precipitation, Reverse Phase HPLC, chromatography on silica, chromatography on heparin SEPHAROSE™, chromatography on an anion or cation exchange resin (such as a polyaspartic acid column), chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are also available depending on the antibody to be recovered.
[0245] Following any preliminary purification step(s), the mixture comprising the antibody of interest and contaminants may be subjected to low pH hydrophobic interaction chromatography using an elution buffer at a pH between, e.g., about 2.5-4.5, and may be performed at low salt concentrations (e.g., from about 0-0.2 5M salt).
VIII. Pharmaceutical formulations, dosing, and routes of administration
[0246] The present disclosure provides antibodies and related compositions, which may be useful for elimination of -expressing pathogens from the body, for example, and for identification and quantification of the number of TMPRSS2-expressing pathogens in biological samples, for example.
[0247] In some embodiments, any of the antibodies or antigen binding fragments thereof may be formulated in a pharmaceutical composition that is useful for a variety of purposes, including the treatment of diseases or disorders. Pharmaceutical compositions comprising one or more antibodies may be administered using a pharmaceutical device to a patient in need thereof, and according to one embodiment of the technology, kits are provided that include such devices. Such devices and kits may be designed for routine administration, including selfadministration, of the pharmaceutical compositions herein.
[0248] Therapeutic formulations of an antibody may be prepared for storage by mixing the agent or antibody having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers (Remington’s Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues ) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionic surfactants such as TWEENTM, PLURONICSTM, or polyethylene glycol (PEG).
[0249] In some embodiments, the disease or disorder is associated with TMPRSS2 expression. In some embodiments, the disease or disorder is associated with aberrant TMPRSS2 expression. In some embodiments, the disease or disorder is associated with Natural Killer (NK), alpha beta T cells, gamma delta T cells, CD8+ T cells, monocytes, or dendritic cells. In some embodiments, the disease or disorder is associated with Natural Killer (NK) cells. In some embodiments, the disease or disorder is associated with alpha beta T cells. In some embodiments, the disease or disorder is associated with gamma delta T cells. In some embodiments, the disease or disorder is associated with CD8+ T cells. In some embodiments, the disease or disorder is associated with monocytes. In some embodiments, the disease or disorder is associated with dendritic cells.
[0250] In some embodiments, the disease or disorder is a cancer, an infectious disease, or an autoimmune disorder. [0251] In some embodiments, the disease or disorder is a cancer. In some embodiments, the cancer is metastatic melanoma, a solid tumor, bladder cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma, hepatic metastasis of colonic origin, papillary thyroid carcinoma, acute myeloid leukemia, or asymptomatic myeloma.
[0252] In some embodiments, the disease or disorder is an infectious disease. In some embodiments, the infectious disease is human immunodeficiency virus (HIV), chronic hepatitis C, cytomegalovirus, or hantavirus.
[0253] In some embodiments, the disease or disorder is an autoimmune disorder. In some embodiments, the autoimmune disorder is Chrohn’s disease, multiple sclerosis, systemic sclerosis, ocular myasthenia gravis, psoriasis or rheumatoid arthritis.
[0254] In some embodiments, any of the antibodies or antigen binding fragments thereof described herein can be used to decrease the production of androgenic hormones in prostate cancer cells.
[0255] In some embodiments, any of the antibodies or antigen binding fragments thereof described herein can be used to inhibit proteolytic cleavage of ACE2 receptor.
[0256] In some embodiments, any of the antibodies or antigen binding fragments thereof described herein can be used to inhibit or reduce cleavage of coronavirus spike glycoproteins. In some embodiments, any of the antibodies or antigen binding fragments thereof described herein can be used to inhibit or reduce viral uptake into a host cell.
[0257] Formulations herein may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Such molecules are suitably present in combination in amounts that are effective for the purpose intended.
[0258] Formulations for in vivo administration generally are sterile. This may be accomplished for instance by filtration through sterile filtration membranes, for example.
[0259] Sustained-release preparations may be prepared. Suitable examples of sustained- release preparations include semipermeable matrices of solid hydrophobic polymers containing the agent/antibody, which matrices are in the form of shaped articles, e.g., films, or microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly (vinyl alcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and gamma ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the Lupron Depot® (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3 -hydroxybutyric acid. While polymers such as such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydrogels release proteins for shorter time periods. When encapsulated agents/antibodies remain in the body for a long time, they may denature or aggregate as a result of exposure to moisture at 37 °C, resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular S-S bond formation through thiol-disulfide interchange, stabilization may be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.
[0260] For therapeutic applications, antibodies provided herein are administered to a mammal, e.g., a human, in a pharmaceutically acceptable dosage form such as those discussed above, including those that may be administered to a human intravenously as a bolus or by continuous infusion over a period of time, or by intramuscular, intraperitoneal, intra- cerebrospinal, subcutaneous, intra-articular, intrasynovial, intrathecal, oral, topical, or inhalation routes. For the prevention or treatment of disease, the appropriate dosage of agent or antibody will depend on the type of disease to be treated, as defined above, the severity and course of the disease, whether the antibody is administered for preventative or therapeutic purposes, previous therapy, the patient’s clinical history and response to the antibody, and the discretion of the attending physician. The antibody is suitably administered to the patient at one time or over a series of treatments.
[0261] Depending on the type and severity of the disease, about 1 pg/kg to about 50 mg/kg (e.g., 0.1-20 mg/kg) of antibody may be an initial candidate dosage for administration to the patient, whether, for example, by one or more separate administrations, or by continuous infusion. A typical daily or weekly dosage might range from about 1 pg/kg to about 20 mg/kg or more, depending on the factors mentioned above. For repeated administrations over several days or longer, depending on the condition, the treatment is repeated until a desired suppression of disease symptoms occurs. However, other dosage regimens may be useful. The progress of this therapy is easily monitored by conventional techniques and assays, including, for example, radiographic imaging. Detection methods using the antibody to determine TMPRSS2 levels in bodily fluids or tissues may be used in order to optimize patient exposure to the therapeutic antibody.
[0262] In some embodiments, a composition comprising an antibody herein can be administered as a monotherapy, and in some embodiments, the composition comprising the antibody can be administered as part of a combination therapy. In some cases, the effectiveness of the antibody in preventing or treating diseases may be improved by administering the antibody serially or in combination with another drug that is effective for those purposes, such as a chemotherapeutic drug for treatment of cancer or a microbial infection. In other cases, the antibody may serve to enhance or sensitize cells to chemotherapeutic treatment, thus permitting efficacy at lower doses and with lower toxicity. Certain combination therapies include, in addition to administration of the composition comprising an antibody that reduces the number of -expressing cells, delivering a second therapeutic regimen selected from the group consisting of a chemotherapeutic agent, radiation therapy, surgery, and a combination of any of the foregoing. Such other agents may be present in the composition being administered or may be administered separately. Also, the antibody may be suitably administered serially or in combination with the other agent or modality, e.g., chemotherapeutic drug or radiation for treatment of cancer, infection, and the like, or an immunosuppressive drug.
IX.. Research and diagnostic
[0263] Also provided herein are diagnostic reagents comprising an antibody described herein. For example, antibodies provided herein may be used to detect and/or purify TMPRSS2 from bodily fluid(s) or tissues. Also provided herein are methods for detecting TMPRSS2. For example, a method may comprise contacting a sample (e.g., a biological sample known or suspected to contain ) with an antibody provided herein, and, if the sample contains TMPRSS2, detecting TMPRSS2: antibody complexes. Also provided herein are reagents comprising an antibody described herein and methods for detecting for research purposes.
[0264] Any of the antibodies or antigen binding fragments disclosed herein can be useful in diagnostic assays for detecting its presence in specific cells, tissues, or bodily fluids. Such diagnostic methods may be useful in diagnosis, e.g., of a hyperproliferative disease or disorder. Thus, clinical diagnostic uses as well as research uses are comprehended herein. In some embodiments, an antibody comprises a detectable marker or label. In some embodiments, an antibody is conjugated to a detectable marker or label. For example, for research and diagnostic applications, an antibody may be labeled with a detectable moiety. Numerous labels are available which are generally grouped into the following categories:
(a) Radioisotopes, such as 35S, 14C, 1251, 3H, and 1311. The antibody can be labeled with the radioisotope using the techniques described in Current Protocols in Immunology, Volumes 1 and 2, Coligen et al., Ed. Wiley-Interscience, New York, N.Y., Pubs. (1991), for example, and radioactivity can be measured using scintillation counting.
(b) Fluorescent labels such as rare earth chelates (europium chelates) or fluorescein and its derivatives, rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin, Texas Red and Brilliant VioletTM are available. The fluorescent labels can be conjugated to the antibody using the techniques disclosed in Current Protocols in Immunology, supra, for example. Fluorescence can be quantified using a flow cytometer, imaging microscope or fluorimeter.
(c) Various enzyme-substrate labels are available. The enzyme generally catalyzes a chemical alteration of the chromogenic substrate that can be measured using various techniques. For example, the enzyme may catalyze a color change in a substrate, which can be measured spectrophotometrically. Alternatively, the enzyme may alter the fluorescence or chemiluminescence of the substrate. Techniques for quantifying a change in fluorescence are described above. The chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light that can be measured (using a chemilluminometer, for example) or donates energy to a fluorescent acceptor. Examples of enzymatic labels include luciferases (e.g., firefly luciferase and bacterial luciferase), luciferin, 2,3- dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, beta-galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclicoxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like. Techniques for conjugating enzymes to antibodies are described in O'Sullivan et al., Methods for the Preparation of Enzyme- Antibody Conjugates for use in Enzyme Immunoassay, in Methods in Enzym. (ed J. Langone & H. Van Vunakis), Academic press, New York, 73: 147-166 (1981). [0265] Examples of enzyme-substrate combinations include, for example:
(i) Horseradish peroxidase (HRP) with hydrogen peroxidase as a substrate, where the hydrogen peroxidase oxidizes a dye precursor (e.g., orthophenylene diamine (OPD) or 3, 3', 5,5'- tetramethyl benzidine hydrochloride (TMB));
(ii) alkaline phosphatase (AP) with para-Nitrophenyl phosphate as chromogenic substrate; and
(iii) P-D -galactosidase (P-D-Gal) with a chromogenic substrate (e.g., p-nitrophenyl-P-D- galactosidase) or fluorogenic substrate 4-methylumbelliferyl-P-D-galactosidase.
[0266] In certain instances, the label is indirectly conjugated with the agent or antibody. The skilled artisan will be aware of various techniques for achieving this. For example, an antibody can be conjugated with biotin and any of the three broad categories of labels mentioned above can be conjugated with avidin, or vice versa. Biotin binds selectively to avidin and thus, the label can be conjugated with the antibody in this indirect manner. Alternatively, to achieve indirect conjugation of the label with the antibody, the antibody is conjugated with a small hapten (e.g., digoxin) and one of the different types of labels mentioned above is conjugated with an anti-hapten antibody (e.g., anti-digoxin antibody). Thus, indirect conjugation of the label with the antibody can be achieved.
[0267] In some embodiments, and antibody or antigen binding fragments thereof need not be labeled, and the presence thereof can be detected, e.g., using a labeled antibody which binds to an antibody.
[0268] In some embodiments, an antibody herein is immobilized on a solid support or substrate. In some embodiments, an antibody herein is non-diffusively immobilized on a solid support (e.g., the antibody does not detach from the solid support). A solid support or substrate can be any physically separable solid to which an antibody can be directly or indirectly attached including, but not limited to, surfaces provided by microarrays and wells, and particles such as beads (e.g., paramagnetic beads, magnetic beads, microbeads, nanobeads), microparticles, and nanoparticles. Solid supports also can include, for example, chips, columns, optical fibers, wipes, filters (e.g., flat surface filters), one or more capillaries, glass and modified or functionalized glass (e.g., controlled-pore glass (CPG)), quartz, mica, diazotized membranes (paper or nylon), polyformaldehyde, cellulose, cellulose acetate, paper, ceramics, metals, metalloids, semiconductive materials, quantum dots, coated beads or particles, other chromatographic materials, magnetic particles; plastics (including acrylics, polystyrene, copolymers of styrene or other materials, polybutylene, polyurethanes, TEFLON™, polyethylene, polypropylene, polyamide, polyester, polyvinylidenedifluoride (PVDF), and the like), polysaccharides, nylon or nitrocellulose, resins, silica or silica-based materials including silicon, silica gel, and modified silicon, Sephadex®, Sepharose®, carbon, metals (e.g., steel, gold, silver, aluminum, silicon and copper), inorganic glasses, conducting polymers (including polymers such as polypyrole and polyindole); micro or nanostructured surfaces such as nucleic acid tiling arrays, nanotube, nanowire, or nanoparticulate decorated surfaces; or porous surfaces or gels such as methacrylates, acrylamides, sugar polymers, cellulose, silicates, or other fibrous or stranded polymers. In some embodiments, the solid support or substrate may be coated using passive or chemically-derivatized coatings with any number of materials, including polymers, such as dextrans, acrylamides, gelatins or agarose. Beads and/or particles may be free or in connection with one another (e.g., sintered). In some embodiments, a solid support or substrate can be a collection of particles. In some embodiments, the particles can comprise silica, and the silica may comprise silica dioxide. In some embodiments the silica can be porous, and in certain embodiments the silica can be non-porous. In some embodiments, the particles further comprise an agent that confers a paramagnetic property to the particles. In certain embodiments, the agent comprises a metal, and in certain embodiments the agent is a metal oxide, (e.g., iron or iron oxides, where the iron oxide contains a mixture of Fe2+ and Fe3+). An antibody may be linked to a solid support by covalent bonds or by non-covalent interactions and may be linked to a solid support directly or indirectly (e.g., via an intermediary agent such as a spacer molecule or biotin).
[0269] Antibodies and antigen binding fragments thereof provided herein may be employed in any known assay method, such as flow cytometry, immunohistochemistry, immunofluorescence, mass cytometry (e.g., Cytof instrument), competitive binding assays, direct and indirect sandwich assays, and immunoprecipitation assays. Zola, Monoclonal Antibodies: A Manual of Techniques, pp. 147-158 (CRC Press, Inc. 1987). Flow cytometry and mass cytometry assays generally involve the use of a single primary antibody to specifically identify the presence of the target molecule expressed on the surface of a dispersed suspension of individual cells. The dispersed cells are typically obtained from a biological fluid sample, e.g., blood, but may also be obtained from a dispersion of single cells prepared from a solid tissue sample such as spleen or tumor biopsy. The primary antibody may be directly conjugated with a detectable moiety, e.g., a fluorophore such as phycoerythrin for flow cytometry or a heavy metal chelate for mass cytometry. Alternatively, the primary antibody may be unlabeled or labeled with an undetectable tag such as biotin, and the primary antibody is then detected by a detectably labeled secondary antibody that specifically recognizes the primary antibody itself or the tag on the primary antibody. The labeled cells are then analyzed in an instrument capable of single cell detection, e.g., flow cytometer, mass cytometer, fluorescence microscope or brightfield light microscope, to identify those individual cells in the dispersed population or tissue sample that express the target recognized by the primary antibody. Detailed description of the technological basis and practical application of flow cytometry principles may be found in, e.g., Shapiro, Practical Flow Cytometry, 4th Edition, Wiley, 2003.
[0270] Sandwich assays involve the use of two antibodies, each capable of binding to a different immunogenic portion, or epitope, of the protein that is detected. In a sandwich assay, the test sample analyte is bound by a first antibody that is immobilized on a solid support, and thereafter a second antibody binds to the analyte, thus forming an insoluble three-part complex. See, e.g., U.S. Pat. No. 4,376,110. The second antibody may itself be labeled with a detectable moiety (direct sandwich assays) or may be measured using an anti-immunoglobulin antibody that is labeled with a detectable moiety (indirect sandwich assay). For example, one type of sandwich assay is an ELISA assay, in which case the detectable moiety is an enzyme. In a cell ELISA, the target cell population may be attached to the solid support using antibodies first attached to the support and that recognize different cell surface proteins. These first antibodies capture the cells to the support. TMPRSS2 on the surface of the cells can then be detected by adding any of the anti-TMPRSS2 antibodies or antigen-binding fragments thereof described herein to the captured cells and detecting the amount of the anti-TMPRSS2 antibody or antigen binding fragment thereof attached to the cells. In certain instances, fixed and permeabilized cells may be used, and in such instances, both surface TMPRSS2 and intracellular TMPRSS2 may be detected.
[0271] In some embodiments, any of the antibodies or antigen binding fragments thereof provided herein are formulated for immunohistochemical analysis. In some embodiments, immunohistochemical analysis includes the use of samples. In some embodiments, immunohistochemical analysis includes the use of blood and/or tissue samples. In some embodiments, the sample may be fresh or frozen or may be embedded in paraffin and fixed with a preservative such as formalin. In some embodiments, the sample is a formalin-fixed paraffin-embedded (FFPE) sample. In some embodiments, the FFPE sample is saturated with formalin (i.e. formaldehyde) and then embedded in a block of paraffin wax. In some embodiments, the FFPE sample is stable at room temperature. In some embodiments, all of the structures in the FFPE sample are preserved. In some embodiments, the intracellular and surface proteins in the FFPE sample are preserved. In some embodiments, the mRNA in the FFPE sample is preserved. In some embodiments, the mRNA, intracellular and surface proteins in the FFPE sample are preserved. In some embodiments, the surface proteins in the FFPE sample are denatured.
[0272] In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting TMPRSS2 in a formalin-fixed paraffin-embedded sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting TMPRSS2 on the surface of a in a formalin-fixed paraffin-embedded sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting intracellular TMPRSS2 in a formalin-fixed paraffin-embedded sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting intracellular TMPRSS2, and TMPRSS2 on the surface of a formalin-fixed paraffin-embedded sample.
[0273] In some embodiments, the sample is a fresh sample that has been frozen. In some embodiments, the sample is a fresh sample that has been cryogenically frozen. In some embodiments, the sample is flash frozen. In some embodiments, the sample if flash frozen and stored at 80°C. In some embodiments, all of the structures in the flash frozen sample are preserved. In some embodiments, the intracellular and surface proteins in the flash frozen sample are preserved. In some embodiments, the mRNA in the flash frozen sample is preserved. In some embodiments, the mRNA, intracellular and surface proteins in the flash frozen sample are preserved. In some embodiments, the surface proteins in the flash frozen sample are denatured.
[0274] In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting TMPRSS2 in a frozen sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting TMPRSS2 on the surface of a frozen sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting intracellular TMPRSS2 in a frozen sample. In some embodiments, any of the anti-TMPRSS2 antibodies or antigen binding fragments thereof provided herein are capable of detecting intracellular TMPRSS2, and TMPRSS2 on the surface of a frozen sample.
[0275] The antibodies herein also may be used for in vivo diagnostic assays. Generally, the antibody is labeled with a radionuclide (such as 11 Un, 99Tc, 14C, 1311, 1251, 3H, 32P, or 35S) so that the bound target molecule can be localized using immunoscintillography.
X. Detection of TMPRSS2
[0276] Provided herein are antibodies and methods for detecting TMPRSS2. In some embodiments, antibodies and methods are provided for detecting TMPRSS2 in a biological sample. In some embodiments, the biological sample is a solid tissue, fluid, or cell. In some embodiments, the TMPRSS2 is detected on the surface of the cell. In some embodiments, the TMPRSS2 is detected intracellularly. In some embodiments, the detection of TMPRSS2 is in vitro. In some embodiments, the detection of TMPRSS2 is in vivo.
[0277] The solid tissue may comprise solid tissue from one or more of adipose tissue, bladder, bone, brain breast cervix, endothelium, gallbladder, kidney, liver, lung, lymph, ovary, prostate, salivary gland, stomach, testis, thyroid, urethra, uterus, vagina, and vulva. In some embodiments, the fluid comprises one or more of amniotic fluid, bile, blood, breast milk, breast fluid, cerebrospinal fluid, lavage fluid, lymphatic fluid, mucous, plasma, saliva, semen, serum, spinal fluid, sputum, tears, umbilical cord blood, urine, and vaginal fluid.
[0278] In some embodiments, the sample comprises immune cells. In some embodiments, the sample comprises a heterogeneous population of immune cells. In some embodiments, the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs).
[0279] In some of any embodiments, any of the antibodies or antigen binding fragments thereof provided herein can be used in the characterization of single cells by measurement of gene-expression levels and cellular proteins. Among such known single cell sequencing platforms suitable for integration with the antibodies or antigen binding fragments thereof described herein is the Drop-seq method, including, but not limited to, microfluidic, plate- based, or microwell, Seq-Well™ method and adaptations of the basic protocol, and InDrop™ method. In another embodiment, a single cell sequencing platform suitable for integration with the antibodies or antigen binding fragments thereof described herein is lOx genomics single cell 3' solution or single cell V(D)J solution, either run on Chromium controller, or dedicated Chromium single cell controller. Other suitable sequencing methods include Wafergen iCell8™ method, Microwell-seq method, Fluidigm CI™ method and equivalent single cell products. Still other known sequencing protocols useful with the antibodies or antigen binding fragments thereof described herein include BD Resolve™ single cell analysis platform and ddSeq (from Illumina® Bio-Rad® SureCell™ WTA 3' Library Prep Kit for the ddSEQ™ System, 2017, Pub. No. 1070-2016-014-B, Illumina Inc., Bio-Rad Laboratories, Inc.). In still other embodiment, the antibodies or antigen binding fragments thereof described herein are useful with combinatorial indexing based approaches (sci-RNA-seq™ method or SPLiT-seq™ method) and Spatial Transcriptomics, or comparable spatially resolved sequencing approaches. The methods and compositions described herein can also be used as an added layer of information on standard index sorting (FACS) and mRNA-sequencing- based approaches.
[0280] In some of any embodiments, any of the antibodies or antigen binding fragments thereof described herein can be used to detect the presence, absence or amount of the various nucleic acids, proteins, targets, oligonucleotides, amplification products and barcodes described herein.
[0281] In some embodiments, the biological sample is from a healthy subject. In some embodiments, the sample is from a subject with a disease or condition. In some embodiments, the detection of TMPRSS2 indicates the presence or absence of a disease or disorder. In some embodiments, the disease or disorder is a cancer, an autoimmune disorder, an inflammatory disorder, a neurologic disorder, or an infection. In some embodiments, the cancer is the cancer is acute myeloid leukemia, acute lymphoblastic leukemia, colorectal, ovarian, gynecologic, liver, glioblastoma, Hodgkin lymphoma, chronic lymphocytic leukemia, esophagus, gastric, pancreas, colon, kidney, head and neck, lung and melanoma.
[0282] In some embodiments, the disease or disorder is associated with TMPRSS2 expression, In some embodiments, the disease or disorder is associated with aberrant TMPRSS2 expression. In some embodiments, the disease or disorder is associated with Natural Killer (NK), alpha beta T cells, gamma delta T cells, CD8+ T cells, monocytes, or dendritic cells. In some embodiments, the disease or disorder is associated with Natural Killer (NK) cells. In some embodiments, the disease or disorder is associated with alpha beta T cells. In some embodiments, the disease or disorder is associated with gamma delta T cells. In some embodiments, the disease or disorder is associated with CD8+ T cells. In some embodiments, the disease or disorder is associated with monocytes. In some embodiments, the disease or disorder is associated with dendritic cells. In some of any embodiments, the disease or disorder is chosen from non-viral cancers, virus-associated cancers, cancers associated with HBV infection, cancers associated with Epstein-Barr virus (EBV) infection, cancers associated with polyomavirus infection, erythema nodosum leprosum (ENL), autoimmune diseases, autoimmune inflammation, autoimmune thyroid diseases, B-cell lymphoma, T-cell lymphoma, acute myeloid leukemia, Hodgkin's Disease, acute myelogenous leukemia, acute myelomonocytic leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, B cell large cell lymphoma, malignant lymphoma, acute leukemia, lymphosarcoma cell leukemia, B-cell leukemias, myelodysplastic syndromes, solid phase cancer, herpes viral infections, and/or rejection of transplanted tissues or organs.
[0283] In some embodiments, the disease or disorder is a cancer, an infectious disease, or an autoimmune disorder.
[0284] In some embodiments, the disease or disorder is a cancer. In some embodiments, the cancer is metastatic melanoma, a solid tumor, bladder cancer, head and neck squamous cell carcinoma, hepatocellular carcinoma, hepatic metastasis of colonic origin, papillary thyroid carcinoma, acute myeloid leukemia, or asymptomatic myeloma.
[0285] In some embodiments, the disease or disorder is an infectious disease. In some embodiments, the infectious disease is human immunodeficiency virus (HIV), chronic hepatitis C, cytomegalovirus, or hantavirus.
[0286] In some embodiments, the disease or disorder is an autoimmune disorder. In some embodiments, the autoimmune disorder is Chrohn’s disease, multiple sclerosis, systemic sclerosis, ocular myasthenia gravis, psoriasis or rheumatoid arthritis. In some embodiments, the autoimmune disorder is Chrohn’s disease, multiple sclerosis, systemic sclerosis, ocular myasthenia gravis, psoriasis or rheumatoid arthritis.
[0287] In some embodiments, any of the antibodies or antigen binding fragments thereof can be used in generating a nucleic acid molecule comprising all or a portion of the sequence of the oligonucleotide or a complement thereof. In some of any embodiments, the antibody or antigen binding fragment thereof can be used in a method of associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
[0288] In some embodiments, any of the antibodies or antigen binding fragments thereof can be used in the construction of a protein library. In some of any embodiments, the construction of a protein library comprises sequencing. In some of any embodiments, the construction of a protein library comprises the use of flow cytometry.
[0289] In some of any embodiments, provided herein is a method of detecting TMPRSS2, comprising a) contacting a sample with the antibody or antigen binding fragment thereof of any of the antibodies or antigen binding fragments thereof under conditions to bind said antibody or antigen binding fragment thereof to a TMPRSS2 receptor on said sample, wherein the binding generates the production of a receptor/antibody or antigen binding fragment thereof of complex; b) detecting the presence of the receptor/antibody or antigen binding fragment thereof of complexes; c) wherein the detecting comprises the presence or absence of the TMPRSS2 receptor on said sample.
[0290] In some of any embodiments, provided herein is a method of treating or preventing a disease or disorder associated with TMPRSS2 in a subject, comprising: a) contacting a sample known or suspected to contain TMPRSS2 with the antibody or antigen binding fragment thereof any of the antibodies or antigen binding fragments thereof, b) detecting the presence of complexes comprising TMPRSS2 and the antibody or antigen binding fragment thereof; wherein the presence of the complexes indicates the presence of a disease or disorder; and c) administering to the subject the antibody or antigen binding fragment thereof of any of the antibodies or antigen binding fragments thereof
[0291] In some of any embodiments, provided herein is a method of diagnosing a disease or disorder, comprising: a) isolating a sample from a subject, b) incubating the sample with the antibody or antigen binding fragment thereof of any of any of the antibodies or antigen binding fragments thereof, for a period of time sufficient to generate TMPRSS2:anti- TMPRSS2 complexes; c) detecting the presence or absence of the TMPRSS2:anti-TMPRSS2 complexes from the isolated tissue, and d) associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
[0292] In some of any embodiments, the increase of TMPRSS2 over a control level in the location of interest of the tissue sample is indicative of a disease or disorder in a subject.
[0293] In some of any embodiments, the detection comprises hybridization of a detectable moiety to the antibody or antigen binding fragment thereof. In some of any embodiments, the sample is contacted with a second antibody. In some of any embodiments, the second antibody is an antibody comprising a detectable moiety. In some of any embodiments, the detectable moiety comprises an oligonucleotide. In some of any embodiments, the detectable moiety comprises a fluorescent label. In some of any embodiments, the measurement comprises sequencing. In some of any embodiments, the detectable moiety comprises immunofluorescence. In some of any embodiments, the sample is a formalin-fixed paraffin- embedded sample. In some of any embodiments, the sample comprises a cell. In some of any embodiments, the sample comprises a tissue sample.
XL Kits incorporating anti- antibodies
[0294] An antibody herein may be provided in a kit, for example, a packaged combination of reagents in predetermined amounts with instructions for use (e.g., instructions for performing a diagnostic assay; instructions for performing a laboratory assay). In some embodiments, the kit is a diagnostic kit configured to detect in a sample (e.g., a biological sample). Where the antibody is labeled with a fluorophore, the kit may include an identical isotype negative control irrelevant antibody to control for non-specific binding of the antibody. Where the antibody is labeled with an enzyme, the kit may include substrates and cofactors required by the enzyme (e.g., substrate precursor which provides the detectable chromophore or fluorophore). Additional additives may be included such as stabilizers, buffers (e.g., a block buffer or lysis buffer), and the like. The relative amounts of the various reagents may be varied widely to provide for concentrations in solution of the reagents that substantially optimize the sensitivity of the assay. In certain instances, reagents may be provided as dry powders (e.g., lyophilized powder), including excipients that on dissolution will provide a reagent solution having the appropriate concentration. EXEMPLARY EMBODIMENTS
[0295] 1. An antibody or antigen binding fragment thereof that binds TMPRSS2 or a portion thereof, comprising: a) an immunoglobulin heavy chain variable domain comprising:
(i) a heavy chain complementary determining region 1 (CDRH1) comprising the sequence X1X2X3X4X5 (SEQ ID NO: 81), wherein Xi is S, N or D; X2 is Y or S; X3 is G, W or D; X4 is V, M or I; and X5 is S, Q or N; and
(ii) a heavy chain complementary determining region 2 (CDRH2) comprising the sequence X1IX2X3X4X5X6X7X8X9X10X11X12X13X14X15X16 (SEQ ID NO: 82), wherein Xi is V, A, Y or W; X2 is W,Y,S or N; X3 is G, P, S or T; X4 is D, V, G or Y; X5 is G, D or T; X6 is S or G; X7 is T, D, R, S or E; Xs is N, T, I or P; X9 is Y, R, K or T; X10 is H, Y or F; Xn is S, T, P, N or A; X12 is A, Q, D or E; X13 is L, K, T or G; Xi4 is I, F or V; X15 is S or K; and Xi6 is G or no amino acid; and
(iii) a heavy chain complementary determining region 3 (CDRH3) comprising the sequence X1X2X3X4X5X6X7X8X9X10X11X12X13 (SEQ ID NO: 83), wherein Xi is P, L, D, A , S or I; X2 is G, S, F, Y or H; X3 is N, P, D, Y or G; X4 is N, S, Y, R, D or G; X5 is Y, N F or no amino acid; Xe is D, Y S or no amino acid; X7 is W, D, Y, H or no amino acid; Xs is Y, F, G, A, W or no amino acid; X9 is F, D, A, M, Y or no amino acid; X10 is D, C or no amino acid; X11 is V, F, D or no amino acid; X12 is D, V or no amino acid; and X13 is Y or no amino acid; and b) an immunoglobulin light chain variable domain comprising:
(i) a light chain complementary determining region 1 (CDRL1) comprising the sequence X1ASX2X3IX4X5X6X7X8 (SEQ ID NO: 84) wherein Xi is K or R; X2 is Q or E; X3 is D, S, E or N; X4 is N, G, S or Y; X5 is K, T, S or V; X6 is Y or N; X7 is I, M or L; and Xs is A, H, S or T; and
(ii) a light chain complementary determining region 2 (CDRL2) comprising the sequence X1X2X3X4X5X6X7 (SEQ ID NO: 85) wherein Xi is Y, R or A; X2 is T, A or G; X3 is S, N or T; X4 is T, E, R or N; X5 is L or S; Xe is Q, I, E, D or A; and X7 is P, S or D; and (iii) a light chain complementary determining region 3 (CDRL3) comprising the sequence X1X2X3X4X5X6X7X8X9 (SEQ ID NO: 86), wherein Xi is L or Q; X2 is Q or H; X3 is Y, S or F; X4 is A, Y, D, H or W; X5 is N, E, S or G; X6 is L, W, P, Y or T; X7 is L or P; Xs is T, L or Y; and X9 is T or no amino acid.
[0296] 2 The antibody or antigen binding fragment thereof of embodiment 1, wherein the immunoglobulin heavy chain variable domain comprises: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 51, 52, 53, 54, 55 or 56; and a CDRH3 comprising the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63.
[0297] 3. The antibody or antigen binding fragment thereof of embodiment 1 or embodiment 2, wherein the immunoglobulin heavy chain variable domain comprises: a CDRH1 comprising the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50; a CDRH2 comprising the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55 or 56; and a CDRH3 comprising the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63.
[0298] 4. The antibody or antigen binding fragment thereof of any of embodiments 1-3, wherein the immunoglobulin light chain variable domain comprises: a CDRL1 comprising the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 70, 71, 72, 73 or 74; and a CDRL3 comprising the sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 75, 76, 77, 78, 79 or 80.
[0299] 5.The antibody or antigen binding fragment thereof of any of embodiments 1-4, wherein the immunoglobulin light chain variable domain comprises: a CDRL1 comprising a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69; a CDRL2 comprising a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74; and a CDRL3 comprising a sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
[0300] 6. The antibody or antigen binding fragment thereof of any of embodiments 1-5, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 45, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45, the CDRH2 comprises the sequence set forth in SEQ ID NO: 51, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 57, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 57.
[0301] 7. The antibody or antigen binding fragment thereof of any of embodiments 1-6, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 45, the CDRH2 comprises the sequence set forth in SEQ ID NO: 51; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 57.
[0302] 8. The antibody or antigen binding fragment thereof of any of embodiments 1-5, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 46, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46; the CDRH2 comprises the sequence set forth in SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:52; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 58.
[0303] 9. The antibody or antigen binding fragment thereof of any of embodiments 1-5 and 8, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 46, the CDRH2 comprises the sequence set forth in SEQ ID NO: 52; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 58.
[0304] 10. The antibody or antigen binding fragment thereof of any of embodiments 1-5, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 47, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47; the CDRH2 comprises the sequence set forth in SEQ ID NO: 53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 58.
[0305] 11. The antibody or antigen binding fragment thereof of any of embodiments 1-5 and 10, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 47, the CDRH2 comprises the sequence set forth in SEQ ID NO: 53; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 58.
[0306] 12. The antibody or antigen binding fragment thereof of any of embodiments 1-5, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 48, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48; the CDRH2 comprises the sequence set forth in SEQ ID NO: 54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:54; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:60.
[0307] 13. The antibody or antigen binding fragment thereof of any of embodiments 1-5 and 12, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 48, the CDRH2 comprises the sequence set forth in SEQ ID NO: 54; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 60.
[0308] 14. The antibody or antigen binding fragment thereof of any of embodiments 1-5, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 45, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45; the CDRH2 comprises the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 61 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:61.
[0309] 15. The antibody or antigen binding fragment thereof of any of embodiments 1-5 and 14, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 45, the CDRH2 comprises the sequence set forth in SEQ ID NO: 51; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 61.
[0310] 16. The antibody or antigen binding fragment thereof of any of embodiments 1-5, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 49, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49; the CDRH2 comprises the sequence set forth in SEQ ID NO: 55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:55; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 62 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,
88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 62.
[0311] 17. The antibody or antigen binding fragment thereof of any of embodiments 1-5 and 16, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 49, the CDRH2 comprises the sequence set forth in SEQ ID NO: 55; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 62.
[0312] 18. The antibody or antigen binding fragment thereof of any of embodiments 1-5, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:50; the CDRH2 comprises the sequence set forth in SEQ ID NO: 56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 63.
[0313] 19. The antibody or antigen binding fragment thereof of any of embodiments 1-5 and 18, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 50, the CDRH2 comprises the sequence set forth in SEQ ID NO: 56; and the CDRH3 comprises the sequence set forth in SEQ ID NO: 63.
[0314] 20. The antibody or antigen binding fragment thereof of any of embodiments 1-19, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 64, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64; the CDRL2 comprises the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 75 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ
ID NO:75.
[0315] 21. The antibody or antigen binding fragment thereof of any of embodiments 1-20, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 64, the CDRL2 comprises the sequence set forth in SEQ ID NO: 70; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 75.
[0316] 22. The antibody or antigen binding fragment thereof of any of embodiments 1-19, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 65 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:65, the CDRL2 comprises the sequence set forth in SEQ ID NO: 71 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:71; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 76 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:76.
[0317] 23. The antibody or antigen binding fragment thereof of any of embodiments 1-15 and 22, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 65, the CDRL2 comprises the sequence set forth in SEQ ID NO: 71; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 76.
[0318] 24. The antibody or antigen binding fragment thereof of any of embodiments 1-19, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 66 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:66, the CDRL2 comprises the sequence set forth in SEQ ID NO: 72 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:72; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 77 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ
ID NO:77.
[0319] 25. The antibody or antigen binding fragment thereof of any of embodiments 1-19 and 24, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 66, the CDRL2 comprises the sequence set forth in SEQ ID NO: 72; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 77.
[0320] 26. The antibody or antigen binding fragment thereof of any of embodiments 1-19, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 67 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:67, the CDRL2 comprises the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:78.
[0321] 27. The antibody or antigen binding fragment thereof of any of embodiments 1-19 and 26, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 67, the CDRL2 comprises the sequence set forth in SEQ ID NO: 73; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 78.
[0322] 28. The antibody or antigen binding fragment thereof of any of embodiments 1-19, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64, the CDRL2 comprises the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 79 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,
I l l 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ
ID NO:79.
[0323] 29. The antibody or antigen binding fragment thereof of any of embodiments 1-19 and 28, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 64, the CDRL2 comprises the sequence set forth in SEQ ID NO: 70; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 79.
[0324] 30. The antibody or antigen binding fragment thereof of any of embodiments 1-19, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 68 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:68, the CDRL2 comprises the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:78.
[0325] 31. The antibody or antigen binding fragment thereof of any of embodiments 1-19 and 30, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 68, the CDRL2 comprises the sequence set forth in SEQ ID NO: 73; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 78.
[0326] 32. The antibody or antigen binding fragment thereof of any of embodiments 1-19, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 69 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:69, the CDRL2 comprises the sequence set forth in SEQ ID NO: 74 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:74; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 80 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ
ID NO:80.
[0327] 33. The antibody or antigen binding fragment thereof of any of embodiments 1-19 and 32, wherein the CDRL1 comprises the sequence set forth in SEQ ID NO: 69, the CDRL2 comprises the sequence set forth in SEQ ID NO: 74; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 80.
[0328] 34. The antibody or antigen binding fragment thereof of any of embodiments 1-33, wherein the CDRH1 comprises the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; the CDRH2 comprises a sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55, or 56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 51, 52, 53, 54, 55, or 56; the CDRH3 comprises a sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63; the CDRL1 comprises a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69; the CDRL2 comprises a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 70, 71, 72, 73 or 74; the CDRL3 comprises a sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 75, 76, 77, 78, 79 or 80.
[0329] 35. The antibody or antigen binding fragment thereof of any of embodiments 1-34, wherein the CDRH1 comprises the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50; the CDRH2 comprises the sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55, or 56; the CDRH3 comprises the sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63; the CDRL1 comprises the sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69; the CDRL2 comprises the sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74; and the CDRL3 comprises the sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80.
[0330] 36. The antibody or antigen binding fragment thereof of any of embodiments 1-35, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45, the CDRH2 comprises the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; the CDRH3 comprises the sequence set forth in SEQ ID NO: 57 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:57; the CDRL1 comprises the sequence set forth in SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64, the CDRL2 comprises the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 75 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:75.
[0331] 37. The antibody or antigen binding fragment thereof of any of embodiments 1-36, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 45, the CDRH2 comprises the sequence set forth in SEQ ID NO: 51; the CDRH3 comprises the sequence set forth in SEQ ID NO: 57; the CDRL1 comprises the sequence set forth in SEQ ID NO: 64, the CDRL2 comprises the sequence set forth in SEQ ID NO: 70; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 75. [0332] 38. The antibody or antigen binding fragment thereof of any of embodiments 1-35, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 46 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:46; the CDRH2 comprises the sequence set forth in SEQ ID NO: 52 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:52; the CDRH3 comprises the sequence set forth in SEQ ID NO: 58 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:58; the CDRL1 comprises the sequence set forth in SEQ ID NO: 65 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:65, the CDRL2 comprises the sequence set forth in SEQ ID NO: 71 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:71; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 76 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:76.
[0333] 39. The antibody or antigen binding fragment thereof of any of embodiments 1-35 and 38, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 46, the CDRH2 comprises the sequence set forth in SEQ ID NO: 52; the CDRH3 comprises the sequence set forth in SEQ ID NO: 58; the CDRL1 comprises the sequence set forth in SEQ ID NO: 65, the CDRL2 comprises the sequence set forth in SEQ ID NO: 71; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 76.
[0334] 40. The antibody or antigen binding fragment thereof of any of embodiments 1-35, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 47 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:47, the CDRH2 comprises the sequence set forth in SEQ ID NO: 53 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:53; the CDRH3 comprises the sequence set forth in SEQ ID NO: 59 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:59; the CDRL1 comprises the sequence set forth in SEQ ID NO: 66 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:66, the CDRL2 comprises the sequence set forth in SEQ ID NO: 72 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:72; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 77 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:77.
[0335] 41. The antibody or antigen binding fragment thereof of any of embodiments 1-35 and 40, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 47, the CDRH2 comprises the sequence set forth in SEQ ID NO: 53; the CDRH3 comprises the sequence set forth in SEQ ID NO: 59; the CDRL1 comprises the sequence set forth in SEQ ID NO: 66, the CDRL2 comprises the sequence set forth in SEQ ID NO: 72; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 77.
[0336] 42. The antibody or antigen binding fragment thereof of any of embodiments 1-35, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 48 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:48, the CDRH2 comprises the sequence set forth in SEQ ID NO: 54 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:54; the CDRH3 comprises the sequence set forth in SEQ ID NO: 60 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:60; the CDRL1 comprises the sequence set forth in SEQ ID NO: 67 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:67, the CDRL2 comprises the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:78.
[0337] 43. The antibody or antigen binding fragment thereof of any of embodiments 1-35 and 42, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 48, the CDRH2 comprises the sequence set forth in SEQ ID NO: 54; the CDRH3 comprises the sequence set forth in SEQ ID NO: 60; the CDRL1 comprises the sequence set forth in SEQ ID NO: 67, the CDRL2 comprises the sequence set forth in SEQ ID NO: 73; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 78.
[0338] 44. The antibody or antigen binding fragment thereof of any of embodiments 1-35, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45, the CDRH2 comprises the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; the CDRH3 comprises the sequence set forth in SEQ ID NO: 61 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:61; the CDRL1 comprises the sequence set forth in SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64, the CDRL2 comprises the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 79 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ
ID NO:79.
[0339] 45. The antibody or antigen binding fragment thereof of any of embodiments 1-35 and 44, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 45, the CDRH2 comprises the sequence set forth in SEQ ID NO: 51; the CDRH3 comprises the sequence set forth in SEQ ID NO: 61; the CDRL1 comprises the sequence set forth in SEQ ID NO: 64, the CDRL2 comprises the sequence set forth in SEQ ID NO: 70; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 79.
[0340] 46. The antibody or antigen binding fragment thereof of any of embodiments 1-35, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 49 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:49, the CDRH2 comprises the sequence set forth in SEQ ID NO: 55 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:55; the CDRH3 comprises the sequence set forth in SEQ ID NO: 62 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:62; the CDRL1 comprises the sequence set forth in SEQ ID NO: 68 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:68, the CDRL2 comprises the sequence set forth in SEQ ID NO: 73 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:73; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 78 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:78.
[0341] 47. The antibody or antigen binding fragment thereof of any of embodiments 1-35 and 46, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 49, the CDRH2 comprises the sequence set forth in SEQ ID NO: 55; the CDRH3 comprises the sequence set forth in SEQ ID NO: 62; the CDRL1 comprises the sequence set forth in SEQ ID NO: 68, the CDRL2 comprises the sequence set forth in SEQ ID NO: 73; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 78.
[0342] 48. The antibody or antigen binding fragment thereof of any of embodiments 1-35, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 50 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:50, the CDRH2 comprises the sequence set forth in SEQ ID NO: 56 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:56; the CDRH3 comprises the sequence set forth in SEQ ID NO: 63 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:63; the CDRL1 comprises the sequence set forth in SEQ ID NO: 69 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:69, the CDRL2 comprises the sequence set forth in SEQ ID NO: 74 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:74; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 80 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:80.
[0343] 49. The antibody or antigen binding fragment thereof of any of embodiments 1-35 and 48, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 50, the CDRH2 comprises the sequence set forth in SEQ ID NO: 56; the CDRH3 comprises the sequence set forth in SEQ ID NO: 63; the CDRL1 comprises the sequence set forth in SEQ ID NO: 69, the CDRL2 comprises the sequence set forth in SEQ ID NO: 74; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 80.
[0344] 50. The antibody or antigen binding fragment thereof of any of embodiments 1-49, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 20-26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20-26.
[0345] 51. The antibody or antigen binding fragment thereof of any of embodiments 1-50, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 20-26.
[0346] 52. The antibody or antigen binding fragment thereof of any of embodiments 1-51, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 20 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:20.
[0347] 53. The antibody or antigen binding fragment thereof of any of embodiments 1-52, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 20.
[0348] 54. The antibody or antigen binding fragment thereof of any of embodiments 1-51, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:21.
[0349] 55. The antibody or antigen binding fragment thereof of any of embodiments 1-51 and 54, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 21.
[0350] 56. The antibody or antigen binding fragment thereof of any of embodiments 1-51, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 22, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:22.
[0351] 57. The antibody or antigen binding fragment thereof of any of embodiments 1-51 and 56, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 22. [0352] 58. The antibody or antigen binding fragment thereof of any of embodiments 1-51, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 23, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:23.
[0353] 59. The antibody or antigen binding fragment thereof of any of embodiments 1-51 and 58, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 23.
[0354] 60. The antibody or antigen binding fragment thereof of any of embodiments 1-51, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 24, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:24.
[0355] 61. The antibody or antigen binding fragment thereof of any of embodiments 1-51 and 60, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 24.
[0356] 62. The antibody or antigen binding fragment thereof of any of embodiments 1-51, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:25.
[0357] 63. The antibody or antigen binding fragment thereof of any of embodiments 1-51 and 62, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 25.
[0358] 64. The antibody or antigen binding fragment thereof of any of embodiments 1-51, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:26. [0359] 65. The antibody or antigen binding fragment thereof of any of embodiments 1-51 and 64, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 26.
[0360] 66. The antibody or antigen binding fragment thereof of any of embodiments 1-65, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 27-33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27-33.
[0361] 67. The antibody or antigen binding fragment thereof of any of embodiments 1-66, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 27-33.
[0362] 68. The antibody or antigen binding fragment thereof of any of embodiments 1-67, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27.
[0363] 69. The antibody or antigen binding fragment thereof of any of embodiments 1-68, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 27.
[0364] 70. The antibody or antigen binding fragment thereof of any of embodiments 1-67, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 28.
[0365] 71. The antibody or antigen binding fragment thereof of any of embodiments 1-67 and 70, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 28.
[0366] 72. The antibody or antigen binding fragment thereof of any of embodiments 1-67, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 29.
[0367] 73. The antibody or antigen binding fragment thereof of any of embodiments 1-67 and 72, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 29.
[0368] 74. The antibody or antigen binding fragment thereof of any of embodiments 1-67, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 30.
[0369] 75. The antibody or antigen binding fragment thereof of any of embodiments 1-67 and 74, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 30.
[0370] 76. The antibody or antigen binding fragment thereof of any of embodiments 1-67, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 31.
[0371] 77. The antibody or antigen binding fragment thereof of any of embodiments 1-67 and 76, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 31.
[0372] 78. The antibody or antigen binding fragment thereof of any of embodiments 1-67, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 32, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 32. [0373] 79. The antibody or antigen binding fragment thereof of any of embodiments 1-67 and 78, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 32.
[0374] 80. The antibody or antigen binding fragment thereof of any of embodiments 1-67, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 33.
[0375] 81. The antibody or antigen binding fragment thereof of any of embodiments 1-67 and 80, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 33.
[0376] 82. The antibody or antigen binding fragment thereof of any of embodiments 1-81, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 20-26 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20-26, and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 27-33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27-33.
[0377] 83. The antibody or antigen binding fragment thereof of any of embodiments 1-82, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 20-26, and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 27-33.
[0378] 84. The antibody or antigen binding fragment thereof of any of embodiments 1-83, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 20, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 20; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27.
[0379] 85. The antibody or antigen binding fragment thereof of any of embodiments 1-84, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 20, and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 27.
[0380] 86. The antibody or antigen binding fragment thereof of any of embodiments 1-83, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 21, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 21; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NOS: 28, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:28.
[0381] 87. The antibody or antigen binding fragment thereof of any of embodiments 1-83 and 86, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 21, and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 28.
[0382] 88. The antibody or antigen binding fragment thereof of any of embodiments 1-83, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 22, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 22; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 29, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:29.
[0383] 89. The antibody or antigen binding fragment thereof of any of embodiments 1-83 and 88, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 22, and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 29.
[0384] 90. The antibody or antigen binding fragment thereof of any of embodiments 1-83, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 23, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 23; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 30, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:30.
[0385] 91. The antibody or antigen binding fragment thereof of any of embodiments 1-83 and 90, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 23, and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 30.
[0386] 92. The antibody or antigen binding fragment thereof of any of embodiments 1-83, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 24, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 24; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 31, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:31.
[0387] 93. The antibody or antigen binding fragment thereof of any of embodiments 1-83 and 92, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 24, and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 31.
[0388] 94. The antibody or antigen binding fragment thereof of any of embodiments 1-83, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 25, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 25; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 32, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:32.
[0389] 95. The antibody or antigen binding fragment thereof of any of embodiments 1-83 and 94, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:25, and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO:32.
[0390] 96. The antibody or antigen binding fragment thereof of any of embodiments 1-83, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 26; and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:33.
[0391] 97. The antibody or antigen binding fragment thereof of any of embodiments 1-83 and 96, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 26, and the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 33.
[0392] 98. The antibody or antigen binding fragment thereof of any of embodiments 1-97, comprising one immunoglobulin heavy chain variable domain and one immunoglobulin light chain variable domain.
[0393] 99. The antibody or antigen binding fragment thereof of any of embodiments 1-98, comprising two immunoglobulin heavy chain variable domains and two immunoglobulin light chain variable domains. [0394] 100. The antibody or antigen binding fragment thereof of any of embodiments 1-99, wherein the antibody or antigen binding fragment thereof is isolated.
[0395] 101. The antibody or antigen binding fragment thereof of any of embodiments 1-
100, wherein the antibody or antigen binding fragment thereof is humanized.
[0396] 102. The antibody or antigen binding fragment thereof of any of embodiments 1-
101, wherein the antibody or antigen binding fragment thereof is conjugated.
[0397] 103. The antibody or antigen binding fragment thereof of any of embodiments 1-
102, further comprising an oligonucleotide.
[0398] 104. The antibody or antigen binding fragment thereof of embodiment 103, wherein the oligonucleotide comprises a sample barcode sequence.
[0399] 105. The antibody or antigen binding fragment thereof of embodiment 103 or embodiment 104, wherein the oligonucleotide comprises a binding site for a primer and an anchor.
[0400] 106. The antibody or antigen binding fragment thereof of any of embodiments 1- 105, wherein the antibody or antigen binding fragment thereof is conjugated to a detectable marker or label.
[0401] 107. The antibody or antigen binding fragment thereof of embodiment 106, wherein the detectable marker or label is conjugated directly to the antigen or antigen binding fragment thereof.
[0402] 108. The antibody or antigen binding fragment thereof of embodiment 106, wherein the detectable marker or label is conjugated to the oligonucleotide.
[0403] 109. The antibody or antigen binding fragment thereof of any of embodiments 106- 108, wherein the detectable marker or label comprises a detectable moiety.
[0404] 110. The antibody or antigen binding fragment thereof of embodiment 109, wherein the detectable moiety is a radioisotope, fluorescent label or enzyme-substrate label. [0405] 111. The antibody or antigen binding fragment thereof of any of embodiments 1- 110, wherein the antibody or antigen binding fragment thereof is non-diffusively immobilized on a solid support.
[0406] 112. The antibody or antigen binding fragment thereof of any of embodiments 1- 111, that is a single chain fragment.
[0407] 113. The antibody or antigen binding fragment thereof of embodiment 112, wherein the single chain fragment is a single chain variable fragment (scFv).
[0408] 114. The antibody or antigen binding fragment thereof of any of embodiments 1-
113, for use in the detection of TMPRSS2 in a sample.
[0409] 115. The antibody or antigen binding fragment thereof of any of embodiments 1-
114, wherein the antibody or antigen binding fragment thereof binds to a cell expressing TMPRSS2 in a sample.
[0410] 116. The antibody or antigen binding fragment thereof of embodiment 114 or embodiment 115, wherein the sample comprises immune cells.
[0411] 117. The antibody or antigen binding fragment thereof of any of embodiments 114- 116, wherein the sample comprises a heterogenous population of immune cells.
[0412] 118. The antibody or antigen binding fragment thereof of embodiment 116 or embodiment 117, wherein the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs).
[0413] 119. The antibody or antigen binding fragment thereof of any of embodiments 114- 118, wherein the sample comprises a cell with a disease or disorder.
[0414] 120. The antibody or antigen binding fragment thereof of embodiment 119, wherein the disease or disorder is a cancer, an autoimmune disorder, an inflammatory disorder, a neurologic disorder, or an infection. [0415] 121. The antibody or antigen binding fragment thereof of embodiment 120, wherein the cancer is acute myeloid leukemia, acute lymphoblastic leukemia, colorectal, ovarian, gynecologic, liver, glioblastoma, Hodgkin lymphoma, chronic lymphocytic leukemia, esophagus, gastric, pancreas, colon, kidney, head and neck, lung and melanoma.
[0416] 122. The antibody or antigen binding fragment thereof of any of embodiments 114-
121, wherein the detection comprises the use of a single antibody or antigen binding fragment thereof to bind a portion of TMPRSS2.
[0417] 123. The antibody or antigen binding fragment thereof of any of embodiments 114-
122, wherein the detection comprises the use of two antibody or antigen binding fragments thereof, each capable of binding to a different portion of TMPRSS2.
[0418] 124. The antibody or antigen binding fragment thereof of any of embodiments 114-
123, wherein the detection of TMPRSS2 is on the surface of a cell.
[0419] 125. The antibody or antigen binding fragment thereof of any of embodiments 114-
124, wherein the detection of TMPRSS2 is intracellular.
[0420] 126. The antibody or antigen binding fragment thereof of any of embodiments 114-
125, wherein the detection of TMPRSS2 indicates the presence or absence of a disease or disorder.
[0421] 127. The antibody or antigen binding fragment thereof of any of embodiments 114-
126, wherein the detection is performed in vitro.
[0422] 128. The antibody or antigen binding fragment thereof of any of embodiments 114- 126, wherein the detection is performed in vivo.
[0423] 129. The antibody or antigen binding fragment thereof of any of embodiments 1- 128, wherein the antibody or antigen binding fragment thereof binds to a TMPRSS2 expressing cell.
[0424] 130. The antibody or antigen binding fragment thereof of embodiment 129, wherein the binding to the TMPRSS2 expressing cell decreases the production of androgenic hormones. [0425] 131. The antibody or antigen binding fragment thereof of embodiment 129, wherein the binding to the TMPRSS2 expressing cell inhibits proteolytic cleavage of ACE2 receptor.
[0426] 132. The antibody or antigen binding fragment thereof of embodiment 129, wherein the binding to the TMPRSS2 expressing cell inhibits or reduces cleavage of coronavirus spike glycoproteins.
[0427] 133. The antibody or antigen binding fragment thereof of embodiment 129, wherein the binding to the TMPRSS2 expressing cell inhibits or reduces viral uptake into a host cell.
[0428] 134. A diagnostic antibody or antigen binding fragment thereof comprising the antibody or antigen binding fragment thereof of any of embodiments 1-133.
[0429] 135. A kit comprising the antibody or antigen binding fragment thereof of any one of embodiments 1-133 or the diagnostic antibody or antigen binding fragment thereof of embodiment 134.
[0430] 136. The kit of embodiment 135, comprising a diagnostics kit configured to detect TMPRSS2 in a biological sample.
[0431] 137. A composition comprising the antibody or antigen binding fragment thereof of any of embodiments 1-133, and a pharmaceutically acceptable excipient.
[0432] 138. The composition of embodiment 137, wherein the antibody or antigen binding fragment thereof of is used as an adjuvant or in conjunction with an adjuvant.
[0433] 139. An isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of the antibody or antigen binding fragment thereof of any of embodiments 1-133.
[0434] 140. The isolated nucleic acid of embodiment 139, wherein the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in any of SEQ ID NOs: 6-12.
[0435] 141. The isolated nucleic acid of embodiment 139 or embodiment 140, wherein the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 6. [0436] 142. The isolated nucleic acid of embodiment 139 or embodiment 140, wherein the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 7.
[0437] 143. The isolated nucleic acid of embodiment 139 or embodiment 140, wherein the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 8.
[0438] 144. The isolated nucleic acid of embodiment 139 or embodiment 140, wherein the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 9.
[0439] 145. The isolated nucleic acid of embodiment 139 or embodiment 140, wherein the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 10.
[0440] 146. The isolated nucleic acid of embodiment 139 or embodiment 140, wherein the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 11.
[0441] 147. The isolated nucleic acid of embodiment 139 or embodiment 140, wherein the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 12.
[0442] 148. An isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any of embodiments 1-133.
[0443] 149. The isolated nucleic acid molecule of embodiment 148, wherein the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in any of SEQ ID NOs: 13-19.
[0444] 150. The isolated nucleic acid of embodiment 148 or embodiment 149, wherein the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 13. [0445] 151. The isolated nucleic acid of embodiment 148 or embodiment 149, wherein the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 14.
[0446] 152. The isolated nucleic acid of embodiment 148 or embodiment 149, wherein the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 15.
[0447] 153. The isolated nucleic acid of embodiment 148 or embodiment 149, wherein the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 16.
[0448] 154. The isolated nucleic acid of embodiment 148 or embodiment 149, wherein the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 17.
[0449] 155. The isolated nucleic acid of embodiment 148 or embodiment 149, wherein the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 18.
[0450] 156. The isolated nucleic acid of embodiment 148 or embodiment 149, wherein the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in SEQ ID NO: 19.
[0451] 157. An isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain and the immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any of embodiments 1-133.
[0452] 158. The isolated nucleic acid of any of embodiments 139-157, wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in any of SEQ ID NOs: 6-12; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in any of SEQ ID NOs: 13-19.
[0453] 159. The isolated nucleic acid of embodiment 158, wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 6; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 13.
[0454] 160. The isolated nucleic acid of embodiment 158, wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 7; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 14.
[0455] 161. The isolated nucleic acid of embodiment 158, wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 8; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 15.
[0456] 162. The isolated nucleic acid of embodiment 158, wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 9; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 16.
[0457] 163. The isolated nucleic acid of embodiment 158, wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 10; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 17.
[0458] 164. The isolated nucleic acid of embodiment 158, wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 11; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 18.
[0459] 165. The isolated nucleic acid of embodiment 158, wherein the nucleotide sequence that encodes the immunoglobulin heavy chain variable domain comprises the sequence set forth in SEQ ID NO: 12; and the immunoglobulin light chain variable domain comprises the sequence of amino acids set forth in SEQ ID NO: 19.
[0460] 166. A recombinant expression vector comprising the isolated nucleic acid of any of embodiments 139- 165. [0461] 167. A recombinant expression vector comprising a first expression cassette and a second expression cassette, wherein the first expression cassette comprises a nucleic acid molecule comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of any one of embodiments 1-133, and the second expression cassette comprises a nucleic acid molecule comprising a nucleotide sequence that encodes an immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any one of embodiments 1-133.
[0462] 168. A recombinant expression vector comprising a first expression cassette and a second expression cassette, wherein the first expression cassette comprises a nucleic acid molecule comprising the nucleotide sequence of any of embodiments 139-165, and the second expression cassette comprises a nucleic acid molecule comprising the nucleotide sequence of any of embodiments 139-165.
[0463] 169. The recombinant expression vector of any of embodiments 166-168, wherein the first and second expression cassettes comprise a promoter.
[0464] 170. A host cell transfected with the recombinant expression vector of any of embodiments 166-169.
[0465] 171. An agent-drag conjugate comprising antibody or antigen binding fragment thereof of any of embodiments 1 -133.
[0466] 172. A composition comprising the antibody-drag conjugate of embodiment 171, and a pharmaceutically acceptable carrier.
[0467] 173. A method of detecting TMPRSS2, comprising a) contacting a sample with the antibody or antigen binding fragment thereof of any of embodiments 1-133, under conditions to bind said antibody or antigen binding fragment thereof to a TMPRSS2 receptor on said sample, wherein the binding generates the production of a receptor/antibody or antigen binding fragment thereof of complex; b) detecting the presence of the receptor/antibody or antigen binding fragment thereof of complexes; c) wherein the detecting comprises the presence or absence of the TMPRSS2 receptor on said sample.
[0468] 174. A method of treating or preventing a disease or disorder associated with TMPRSS2 in a subject, comprising: a) contacting a sample known or suspected to contain TMPRSS2 with the antibody or antigen binding fragment thereof of any of embodiments 1-133, b) detecting the presence of complexes comprising TMPRSS2 and the antibody or antigen binding fragment thereof; wherein the presence of the complexes indicates the presence of a disease or disorder; and c) administering to the subject the antibody or antigen binding fragment thereof of any of embodiments 1-133.
[0469] 175. A method of diagnosing a disease or disorder, comprising: a) isolating a sample from a subject b) incubating the sample with the antibody or antigen binding fragment thereof of any of embodiments 1-133, for a period of time sufficient to generate TMPRSS2:anti- TMPRSS2 complexes; c) detecting the presence or absence of the TMPRSS2:anti-TMPRSS2 complexes from the isolated tissue, and d) associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
[0470] 176. The method of embodiment 175, wherein the increase of TMPRSS2 over a control level in the location of interest of the tissue sample is indicative of a disease or disorder in a subject.
[0471] 177 .The method of any of embodiments 173-176, wherein the method is performed in vitro. [0472] 178. The method of any of embodiments 173-176, wherein the method is performed in vivo.
[0473] 179. The method of any of embodiments 173-178, wherein the detection comprises intracellular detection.
[0474] 180. The method of any of embodiments 173-179, wherein the detection comprises detection on the surface of a cell.
[0475] 181. The method of any of embodiments 173-180, wherein the detection comprises hybridization of a detectable moiety to the antibody or antigen binding fragment thereof.
[0476] 182. The method of any of embodiments 173-181, wherein the sample is contacted with a second antibody.
[0477] 183. The method of any of embodiments 173-182, wherein the second antibody is an antibody comprising a detectable moiety.
[0478] 184. The method of any of embodiments 173-183, wherein the detectable moiety comprises an oligonucleotide.
[0479] 185. The method of any of embodiments 173-184, wherein the detectable moiety comprises a fluorescent label.
[0480] 186. The method of any of embodiments 173-185, wherein the measurement comprises sequencing.
[0481] 187. The method of any of embodiments 173-186, wherein the detectable moiety comprises immunofluorescence.
[0482] 188. The method of any of embodiments 173-187, wherein sample is a formalin-fixed paraffin-embedded sample.
[0483] 189. The method of any of embodiments 173-188, wherein the sample comprises a cell.
[0484] 190. The method of any of embodiments 173-189, wherein the sample comprises a tissue sample. [0485] 191. The method of any of embodiments 173-190, wherein the sample comprises immune cells.
[0486] 192. The method of embodiment 191, wherein the immune cell is selected from B cells, plasmacytoid dendritic cells (pDCs), lymphocytes, leukocytes, T cells, monocytes, macrophages, neutrophils, myeloid dendritic cells (mDCs), innate lymphoid cells, mast cells, eosinophils, basophils, natural killer cells, and peripheral blood mononuclear cells (PBMCs).
[0487] 193. The method of any of embodiments 173-192, wherein the sample comprises a tissue or cells associated with a disease or disorder.
[0488] 194. The method of any of embodiments 173-193, wherein the disease or disorder is a cancer, an autoimmune disorder, an inflammatory disorder, or an infection.
[0489] 195. The method of embodiment 193 or embodiment 194, wherein the disease or disorder is chosen from non-viral cancers, virus-associated cancers, cancers associated with HBV infection, cancers associated with Epstein-Barr virus (EBV) infection, cancers associated with polyomavirus infection, erythema nodosum leprosum (ENL), autoimmune diseases, autoimmune inflammation, autoimmune thyroid diseases, B-cell lymphoma, T-cell lymphoma, acute myeloid leukemia, Hodgkin's Disease, acute myelogenous leukemia, acute myelomonocytic leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, B cell large cell lymphoma, malignant lymphoma, acute leukemia, lymphosarcoma cell leukemia, B-cell leukemias, myelodysplastic syndromes, solid phase cancer, herpes viral infections, and/or rejection of transplanted tissues or organs.
[0490] 196. The antibody or antigen binding fragment thereof of any of embodiments 1- 133, for use in a method of associating presence or abundance of TMPRSS2 with a location of interest of a tissue sample.
[0491] 197. The antibody or antigen binding fragment thereof of any of embodiments 1- 133, for use in a method of detecting TMPRSS2 in a tissue sample.
[0492] 198. The antibody or antigen binding fragment thereof of embodiments 196 or embodiment 197, wherein the method comprises generating a nucleic acid molecule comprising all or a portion of the sequence of the oligonucleotide or a complement thereof. [0493] 199. The antibody or antigen binding fragment thereof of any of embodiments 1- 133, for use in the construction of a protein library.
[0494] 200. The antibody or antigen binding fragment thereof of embodiment 199, wherein the construction of a protein library comprises sequencing.
[0495] 201. The antibody or antigen binding fragment thereof of embodiment 199, wherein the construction of a protein library comprises the use of flow cytometry.
EXAMPLES
EXAMPLE 1. Generation anti-TMPRSS2 antibody expressing hybridomas.
[0496] This Example describes the generation and characterization of hybridomas that secrete monoclonal antibodies that react with TMPRSS2 using murine models.
[0497] Briefly, mice were immunized with a TMPRSS2 immunogen, and hybridomas were formed using standard protocols to fuse myeloma cells with spleens, and lymph node cells were drained and harvested. Successful fusions were selected into HAT medium, and cloned into approximately one cell per well in microtiter plates, after which culture supernatants were tested against TMPRSS2-expressing cell transfectants by flow cytometry. Wells were selected by assessment of staining profiles and then sub-cultured into larger vessels and subcloned. Hybridoma sub-clones were further characterized by flow cytometry using TMPRSS2-transfected cells. Candidate clones expressing exemplary anti-TMPRSS2 antibodies were selected and screened using various methods, including by flow cytometry against human blood cells divided into distinct subsets (e.g., lymphocytes, monocytes, and the like), and against one or more cell lines generated from diseased and/or infected human cells. The percentage of positive cells in each blood cell subset was quantified as compared to isotype control.
EXAMPLE 2. Sequencing of exemplary anti-TMPRSS2 antibody variable regions.
[0498] This Example describes the sequencing of exemplary anti-TMPRSS2 antibodies generated in Example 1 above. [0499] Cells from anti-TMPRSS2 hybridoma cell lines described in Example 1 above were grown in standard mammalian tissue culture media. Total RNA was isolated from hybridoma cells from various clones expressing anti-TMPRSS2 monoclonal antibodies using a procedure based on the RNeasy Mini Kit (Qiagen). Briefly, RNA was used to generate a first strand of cDNA of the light chain and heavy chain variable domains. Both light chain and heavy chain variable domain cDNAs were amplified by a 5 ’-RACE technique, and positive clones were prepared by PCR and then subjected to DNA sequencing.
[0500] Amino acid sequences of the individual variable domains (CDRs and Framework regions), including the CDR1, CDR2, and CDR3 regions, for both the heavy and light chains for seven different antibodies (clones), designated AB 1-7 (also referred to herein as antibodies 1-7, and clones 1-7), are shown in FIGs. 1 A and IB. The various heavy and light chain CDR sequences are shown in Table El, below.
Figure imgf000142_0001
Figure imgf000143_0001
Figure imgf000144_0001
EXAMPLE 3. Detection of TMPRSS2 expressing cells using exemplary anti-TMPRSS2 antibodies.
[0501] This Example describes the ability of exemplary generated anti-TMPRSS2 antibodies to detect cells expressing TMPRSS2 by flow cytometry and immunohi stochemi stry . [0502] In a first experiment, exemplary anti-TMPRSS2 antibodies were assessed on cells from an immortalized human colorectal adenocarcinoma cell line (Caco-2 cells; (ATCC® HTB-37).
[0503] Caco-2 cells were grown in DMEM media supplemented with 10% FBS in T75 culture flask, to about 80% confluency. Once cells reached 80% confluency, cells where dislodged from the flask using accutase and suspended in cell staining buffer. 2pg of exemplary generated anti-TMPRSS2 antibodies were added, and allowed to incubate for 15 minutes. Cells were then washed twice with FACS wash buffer and stained with anti-mouse IgG-PE secondary antibody for 15 minutes. Cells where washed with FACS buffer and analyzed on a BD LSRII flow cytometer. A commercially available antibody was used as positive control. As shown in FIGs. 2A-2E, exemplary tested anti-TMPRSS2 antibodies AB1, AB2, AB4, and AB6 (FIGs. 2A-2D) demonstrated similar staining profiles to a commercial antibody (CA; FIG. 2E), compared to isotype control, on Caco-2 cells.
[0504] Exemplary antibodies were further assessed on non-TMPRSS2 expressing HELA cells (ATCC® CCL-2) and white blood cells (lymphocytes, monocytes and granulocytes) isolated from healthy volunteer donors. Briefly, HeLa cells were grown in DMEM media supplemented with 10% FBS in T75 culture flask, to about 80% confluency. White blood cells were stained in whole blood followed by red blood cell lysis. Lysed blood was washed twice with FACS wash buffer and stained with anti-mouse IgG-PE secondary antibody for 15 minutes. Cells where washed with FACS buffer and analyzed on a BD LSRII flow cytometer. As shown in FIG. 3 A-3F and FIGs. 4A-4C, exemplary tested antibodies did not show surface staining on HELA cells (FIG. 3 A-3F), lymphocytes (FIG. 4A), monocytes (FIG. 4B), or granulocytes (FIG. 4C).
[0505] These results demonstrate the ability of exemplary generated anti-TMPRSS2 antibodies to specifically recognize cells expressing the cognate receptor TMPRSS2, with minimal to no non-specific binding.
[0506] In a second experiment, the ability of exemplary generated anti-TMPRSS2 antibodies to stain formalin-fixed paraffin-embedded (FFPE) samples was assessed by immunohistochemistry. 5 pm sections of FFPE samples of human kidney and lymph node were deparaffmized using xylene, and rehydrated in graded ethanol. Heat mediated antigen retrieval was performed using Sodium Citrate pH 6.0 at 90C for 30 minutes. Samples were permeabilized with 0.1% Triton X-100 in PBS for 30 minutes and blocked with 5% FBS in PBS for 1 h. Samples were stained with 5pg/ml of purified exemplary anti-human TMPRSS2 antibody AB1, AB2, AB3, AB5 or AB7 over night at 4C. Cells were then washed and stained with 2.5ug/ml of Alexa-555 conjugated anti-mouse IgG for 1 hour at room temperature in the dark, followed by two washes in PBS. Samples were mounted using Antifade gold with DAPI and imaged using with Metamorph software and analyzed using image J.
[0507] As shown in Fig. 5A, exemplary generated anti-TMPRSS2 antibodies AB1, AB2, AB3, AB5 and AB7 are capable of staining TMPRSS2 expressing human kidney paraffin sections. As shown in FIG. 5B, exemplary tested antibodies AB1, AB2, AB5 and AB7 did not result in staining on non-TMPRSS2 expressing lymph node sections.
[0508] These results demonstrate the ability of exemplary generated anti-TMPRSS2 antibodies to specifically bind the cognate receptor TMPRSS2, with minimal to no nonspecific binding.
EXAMPLE 4. Assessment of antibody blocking ability of exemplary anti-TMPRSS2 antibodies.
[0509] This Example describes the ability of exemplary generated anti-TMPRSS2 antibodies to block binding of other anti-TMPRSS2 antibodies.
[0510] For blocking studies, 100 pL of Caco-2 cell suspension were incubated with 10 pg purified isotype control or AB1, AB2, AB4 and AB6. After 15 minutes, a commercially available (CA) antibody was added for another 15 minutes. Cells were washed 2 times with Cell Staining Buffer, followed by PE-conjugated anti-rabbit IgG. Cells were washed and acquired in a LSRII flow cytometer (BD Biosciences); the data was analyzed using FlowJo software.
[0511] Percentage original MFI was calculated by dividing the MFI of samples blocked with AB1, AB2, AB4 or AB6 by the MFI of samples blocked with the corresponding isotype control, as shown in Table E2. This value was subtracted from 100 to get a blocking percentage. The formula is shown below: n / . . „ > > , [MFI blocking] .
% Blocking
& = 100 - * 100) [MFI no blocking] 7
Figure imgf000147_0001
[0512] As shown in Table E2, exemplary anti-TMPRSS2 antibodies AB1, AB2, AB4 and AB6 are capable of blocking binding of an exemplary tested commercial antibody. These results suggest that exemplary anti-TMPRSS2 antibodies AB1, AB2, AB4 and AB6 are capable of recognizing similar epitopes.
EXAMPLE 5. Assessment of functional activity of exemplary anti-TMPRSS2 antibodies.
[0513] This Example describes the functional assessment of exemplary anti-TMPRSS2 antibodies as measured by the inhibition of TMPRSS2 protease activity and their effect on cell migration, as compared to both commercially available antibodies and the broad spectrum serine protease inhibitor Nafamostat, which has been shown to inhibit TMPRSS2 activity.
[0514] Whole blood from healthy volunteer donors was obtained by venous puncture into vials containing EDTA and fractionated into platelet rich plasma (PRP) by centrifugation for 15 minutes at 800rpm. Platelet rich-plasma was transferred to a fresh tube, diluted 1/3 with PBS. 25 pl of platelets in PBS were added to each well of a 96 well plate, followed by addition lOOpl of diluted AB 1 at a final concentration of 2, 1, 0.5 0.25 pg per well. The broad protease inhibitor Nafamostat Mesylate at final concentrations of 50pM, 25 pM, 12 pM and 6 pM and untreated platelets (platelets and substrate only) were used as positive and negative controls. Boc-Gln-Ala-ArgAMC, a protease substrate that exhibits fluorescence when cleaved by proteases, was added at a final concentration of 5pM. Fluorescence was measured after 3h at 37C, and protease activity was normalized to untreated controls (platelets and substrate only). As shown in Fig. 6A, AB1 showed inhibition of protease activity at multiple concentrations tested compared to control.
[0515] In a similar experiment, recombinant TMPRSS2 was incubated with AB 1 diluted at a final concentration of 2, 1, 0.5, 0.25, 0.125 and 0.6 pg per well. The broad protease inhibitor Nafamostat Mesylate at final concentrations of 50pM, 25 pM, 12 pM and 6 pM and untreated platelets (platelets and substrate only) and untreated recombinant TMPRSS2 used as controls. As shown in FIG. 6B, AB1 showed inhibition of protease activity at multiple concentrations tested compared to Nafamostat and control.
[0516] In another experiment, the effect of anti-TMPRSS2 antibodies on the ability of cells to migrate was tested by a Matrigel invasion assay. The human colon-carcinoma cell line (Caco-2) was serum starved for 24h, after which 5xl05 cells were resuspended in serum-free DMEM media with AB 1 at 40, 20 and lOug/ml, Nafamostat (25pg/mL), with isotype control antibody (20pg/mL) and untreated cells used as controls, and seeded onto the top well of Corning BioCoat Matrigel Invasion Chamber. DMEM media with 20% serum was placed in the bottom well as chemoattractant. After 40h, cells were removed from the top well with a cotton swab and cells that had migrated to the bottom of the membrane were counted. Each experiment was performed in duplicate and 3 images were counted using 91 Oxo objective for each condition using the cell counter feature in Image J software. As shown in Figure 6C, AB1 inhibited the ability of Caco-2 cells to invade the Matrigel and migrate to the bottom of the membrane at 40, 20 and lOug/ml. In a similar experiment, cells were resuspended in serum-free DMEM media with AB1, Nafamostat or control antibodies CA-1-CA4, all at 20 ug/ml, with isotype used as a control, and the assay carried out similar to above. As shown in Figure 6D, AB1 inhibited the ability of Caco-2 cells to invade the Matrigel and migrate to the bottom of the membrane to a higher extent than any commercially tested antibody, and to a degree similar to a broad spectrum serine protease inhibitor (Nafamostat).
[0517] These results suggest that the exemplary generated anti-TMPRSS2 antibody AB1 is capable of inhibiting the functional activity of TMPRSS2 with stronger blocking activity than any tested commercially available antibody. In addition, the degree of inhibition was comparable to the potent inhibitory effects of the serine protease inhibitor Nafamostat. However, while Nafamostat has additional inhibitory effects on multiple types of serine proteases given it’s broad spectrum activity, anti-TMPRSS2 antibodies have the potential for more specific activity and less off target effects, providing potential advantages when used for TMPRSS2 specific purposes.
[0518] The above examples are provided to illustrate the disclosure but not to limit its scope. Other variants of the disclosure will be readily apparent to one of ordinary skill in the art and are encompassed by the appended claims. All publications, databases, internet sources, patents, patent applications, and accession numbers cited herein are hereby incorporated by reference in their entireties for all purposes.
SEQUENCE TABLE
Figure imgf000150_0001
Figure imgf000151_0001
Figure imgf000152_0001
Figure imgf000153_0001
Figure imgf000154_0001
Figure imgf000155_0001
Figure imgf000156_0001
Figure imgf000157_0001
Figure imgf000158_0001
Figure imgf000159_0001

Claims

WHAT IS CLAIMED IS:
1. An antibody or antigen binding fragment thereof that binds TMPRSS2 or a portion thereof, comprising: a) an immunoglobulin heavy chain variable domain comprising:
(i) a heavy chain complementary determining region 1 (CDRH1) comprising the sequence X1X2X3X4X5 (SEQ ID NO: 81), wherein Xi is S, N or D; X2 is Y or S; X3 is G, W or D; X4 is V, M or I; and X5 is S, Q or N; and
(ii) a heavy chain complementary determining region 2 (CDRH2) comprising the sequence X1IX2X3X4X5X6X7X8X9X10X11X12X13X14X15X16 (SEQ ID NO: 82), wherein Xi is V, A, Y or W; X2 is W,Y,S or N; X3 is G, P, S or T; X4 is D, V, G or Y; X5 is G, D or T; X6 is S or G; X7 is T, D, R, S or E; Xs is N, T, I or P; X9 is Y, R, K or T; X10 is H, Y or F; Xn is S, T, P, N or A; X12 is A, Q, D or E; X13 is L, K, T or G; Xi4 is I, F or V; X15 is S or K; and Xi6 is G or no amino acid; and
(iii) a heavy chain complementary determining region 3 (CDRH3) comprising the sequence X1X2X3X4X5X6X7X8X9X10X11X12X13 (SEQ ID NO: 83), wherein Xi is P, L, D, A , S or I; X2 is G, S, F, Y or H; X3 is N, P, D, Y or G; X4 is N, S, Y, R, D or G; X5 is Y, N F or no amino acid; Xe is D, Y S or no amino acid; X7 is W, D, Y, H or no amino acid; Xs is Y, F, G, A, W or no amino acid; X9 is F, D, A, M, Y or no amino acid; X10 is D, C or no amino acid; X11 is V, F, D or no amino acid; X12 is D, V or no amino acid; and X13 is Y or no amino acid; and b) an immunoglobulin light chain variable domain comprising:
(i) a light chain complementary determining region 1 (CDRL1) comprising the sequence X1ASX2X3IX4X5X6X7X8 (SEQ ID NO: 84) wherein Xi is K or R; X2 is Q or E; X3 is D, S, E or N; X4 is N, G, S or Y; X5 is K, T, S or V; X6 is Y or N; X7 is I, M or L; and Xs is A, H, S or T; and
(ii) a light chain complementary determining region 2 (CDRL2) comprising the sequence X1X2X3X4X5X6X7 (SEQ ID NO: 85) wherein Xi is Y, R or A; X2 is T, A or G; X3 is S, N or T; X4 is T, E, R or N; X5 is L or S; Xe is Q, I, E, D or A; and X7 is P, S or D; and (iii) a light chain complementary determining region 3 (CDRL3) comprising the sequence X1X2X3X4X5X6X7X8X9 (SEQ ID NO: 86), wherein Xi is L or Q; X2 is Q or H; X3 is Y, S or F; X4 is A, Y, D, H or W; X5 is N, E, S or G; X6 is L, W, P, Y or T; X7 is L or P; Xs is T, L or Y; and X9 is T or no amino acid.
2. The antibody or antigen binding fragment thereof of claim 1, wherein the CDRH1 comprises the sequence of amino acids set forth in SEQ ID NO: 45, 46, 47, 48, 49 or 50, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 45, 46, 47, 48, 49 or 50; the CDRH2 comprises a sequence of amino acids set forth in SEQ ID NO: 51, 52, 53, 54, 55, or 56, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 51, 52, 53, 54, 55, or 56; the CDRH3 comprises a sequence of amino acids set forth in SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 57, 58, 59, 60, 61, 62 or 63; the CDRL1 comprises a sequence of amino acids set forth in SEQ ID NO: 64, 65, 66, 67, 68 or 69, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 64, 65, 66, 67, 68 or 69; the CDRL2 comprises a sequence of amino acids set forth in SEQ ID NO: 70, 71, 72, 73 or 74, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 70, 71, 72, 73 or 74; the CDRL3 comprises a sequence of amino acids set forth in SEQ ID NO: 75, 76, 77, 78, 79 or 80, or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO: 75, 76, 77, 78, 79 or 80.
3. The antibody or antigen binding fragment thereof claim 1 or claim 2, wherein the CDRH1 comprises the sequence set forth in SEQ ID NO: 45 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:45, the CDRH2 comprises the sequence set forth in SEQ ID NO: 51 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:51; the CDRH3 comprises the sequence set forth in SEQ ID NO: 57 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:57; the CDRL1 comprises the sequence set forth in SEQ ID NO: 64 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:64, the CDRL2 comprises the sequence set forth in SEQ ID NO: 70 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:70; and the CDRL3 comprises the sequence set forth in SEQ ID NO: 75 or a sequence of amino acids that exhibits at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to SEQ ID NO:75.
4. The antibody or antigen binding fragment thereof of any of claims 1-3, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 20-26, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 20-26.
5. The antibody or antigen binding fragment thereof of any of claims 1-4, wherein the immunoglobulin heavy chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 20 or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:20.
6. The antibody or antigen binding fragment thereof of any of claims 1-5, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in any of SEQ ID NOS: 27-33, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to any of SEQ ID NOS: 27-33.
160
7. The antibody or antigen binding fragment thereof of any of claims 1-6, wherein the immunoglobulin light chain variable domain comprises the amino acid sequence set forth in SEQ ID NO: 27, or a sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO:27.
8. The antibody or antigen binding fragment thereof of any of claims 1-7, comprising two immunoglobulin heavy chain variable domains and two immunoglobulin light chain variable domains.
9. The antibody or antigen binding fragment thereof of any of claims 1-8, further comprising one or more human framework regions.
10. The antibody or antigen binding fragment thereof of any of claims 1-9, wherein the antibody or antigen binding fragment thereof is humanized.
11. The antibody or antigen binding fragment thereof of any of claims 1-10, wherein the antibody or antigen binding fragment thereof is conjugated to a detectable marker or label.
12. The antibody or antigen binding fragment thereof of any of claims 1-11, further comprising an oligonucleotide.
13. The antibody or antigen binding fragment thereof of any of claims 1-12, wherein the antibody or antigen binding fragment thereof is non-diffusively immobilized on a solid support.
14. The antibody or antigen binding fragment thereof of any of claims 1-13, that is a single chain fragment.
15. The antibody or antigen binding fragment thereof of claim 14, wherein the single chain fragment is a single chain variable fragment (scFv).
16. A composition comprising the antibody or antigen binding fragment thereof of any of claims 1-15, and a pharmaceutically acceptable excipient.
161
17. The composition of claim 16, wherein the antibody or antigen binding fragment thereof of is used as an adjuvant or in conjunction with an adjuvant.
18. An isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of the antibody or antigen binding fragment thereof of any of claims 1-15.
19. The isolated nucleic acid of claim 18, wherein the immunoglobulin heavy chain variable domain comprises the sequence of nucleotides set forth in any of SEQ ID NOs: 6-12.
20. An isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any of claims 1-15.
21. The isolated nucleic acid molecule of claim 20, wherein the immunoglobulin light chain variable domain comprises the sequence of nucleotides set forth in any of SEQ ID NOs: 13-19.
22. An isolated nucleic acid comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain and the immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any of claims 1-15.
23. A recombinant expression vector comprising the isolated nucleic acid of any of claims 18-22.
24. A recombinant expression vector comprising a first expression cassette and a second expression cassette, wherein the first expression cassette comprises a nucleic acid molecule comprising a nucleotide sequence that encodes the immunoglobulin heavy chain variable domain of any one of claims 1-15, and the second expression cassette comprises a nucleic acid molecule comprising a nucleotide sequence that encodes an immunoglobulin light chain variable domain of the antibody or antigen binding fragment thereof of any one of claims 1-15.
25. The recombinant expression vector of claim 24, wherein the first and second expression cassettes comprise a promoter.
162
26. A host cell transfected with the recombinant expression vector of any of claims
23-25.
27. A method of detecting TMPRSS2, comprising a) contacting a sample with the antibody or antigen binding fragment thereof of any of claims 1-15, under conditions to bind said antibody or antigen binding fragment thereof to a TMPRSS2 receptor on said sample, wherein the binding generates the production of a receptor/antibody or antigen binding fragment thereof of complex; b) detecting the presence of the receptor/antibody or antigen binding fragment thereof of complexes; c) wherein the detecting comprises the presence or absence of the TMPRSS2 receptor on said sample.
28 .The method of claim 27, wherein the method is performed in vitro.
29. The method of claim 27 or claim 28, wherein the detection comprises intracellular detection.
30. The method of claim 27 or claim 28, wherein the detection comprises detection on the surface of a cell.
PCT/US2023/060179 2022-01-07 2023-01-05 Tmprss2 binding antibodies and antigen binding fragments thereof WO2023133470A2 (en)

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