WO2016073906A2 - Immunoessais liés à des facteurs de croissance transformants - Google Patents

Immunoessais liés à des facteurs de croissance transformants Download PDF

Info

Publication number
WO2016073906A2
WO2016073906A2 PCT/US2015/059557 US2015059557W WO2016073906A2 WO 2016073906 A2 WO2016073906 A2 WO 2016073906A2 US 2015059557 W US2015059557 W US 2015059557W WO 2016073906 A2 WO2016073906 A2 WO 2016073906A2
Authority
WO
WIPO (PCT)
Prior art keywords
amino acid
acid sequence
set forth
seq
human
Prior art date
Application number
PCT/US2015/059557
Other languages
English (en)
Other versions
WO2016073906A3 (fr
Inventor
Michelle STRAUB
Dong Yun Lee
William K. MCCONAUGHY
Jared J. SHEEHAN
Katherine Jane Turner
Nagesh K. Mahanthappa
Gregory J. Carven
Justin W. JACKSON
Original Assignee
Scholar Rock, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Scholar Rock, Inc. filed Critical Scholar Rock, Inc.
Publication of WO2016073906A2 publication Critical patent/WO2016073906A2/fr
Publication of WO2016073906A3 publication Critical patent/WO2016073906A3/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/22—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against growth factors ; against growth regulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
    • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Definitions

  • Embodiments of the present disclosure may include modulators of growth factor activity.
  • such modulators may include antibodies and may modulate TGF- ⁇ family member activity and/or biology.
  • Cell signaling molecules stimulate a variety of cellular activities. Such signaling is often tightly regulated, often through interactions with other biomolecules, the extracellular and/or cellular matrix or within a particular cell environment or niche. Such interactions may be direct or indirect.
  • Cell signaling cascades are involved in a number of diverse biological pathways including, but not limited to modulation of cell growth, modulation of tissue homeostasis, extracellular matrix (ECM) dynamics, modulation of cell migration, invasion and immune modulation/suppression.
  • ECM extracellular matrix
  • proteins involved in cell signaling are synthesized and/or are sequestered in latent form, requiring stimulus of some kind to participate in signaling events.
  • antibodies have selected binding profiles to TGFP family member proteins and/or forms thereof.
  • antibodies with selected binding profiles specifically bind to different forms of TGFP family member proteins.
  • antibodies provided herein specifically bind to one or more of a pro-form, a latent-form, a primed-form, and/or a mature- form of a TGFP family member protein, such as proMyostatin, latent Myostatin, primed Myostatin and/or mature Myostatin.
  • proMyostatin proMyostatin
  • latent Myostatin latent Myostatin
  • primed Myostatin primed Myostatin
  • mature Myostatin such antibodies are useful for diagnostic and/or research tools.
  • antibodies provided herein may be useful for detecting the level of one or more forms of a TGFP family member protein in the plasma of a subject using, for example, an enzyme-linked immunosorbent assay (ELISA).
  • ELISA enzyme-linked immunosorbent assay
  • antibodies provided herein may be useful for determining the level and/or localization of one or more forms of a TGFP family member protein in a tissue sample (e.g., a muscle biopsy) using immunohistochemical techniques.
  • tissue sample e.g., a muscle biopsy
  • Such antibodies may be useful for diagnosing diseases/disorders, determining prognosis, and/or assessing response to treatments.
  • aspects of the disclosure relate to a method for assessing a biological sample, which includes obtaining one or more antibodies, each antibody having a selected Myostatin- related binding profile and subjecting the biological sample to an immunoassay using the one or more antibodies.
  • the disclosure relates to antibodies having a selected myo statin-related binding profile, which comprises a set of one or more parameters indicative of the extent to which an antibody specifically binds to one or more antigens, at least one of which antigens is a myostatin-related protein (e.g., Myostatin, GDF11, etc.).
  • the Myostatin-related binding profile is selected from table 33.
  • a Myostatin-related binding profile is referred to herein as a Myostatin binding profile.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin, human GDF-8 prodomain, human GDF- 11 ARM8 prodomain, and human proGDFl 1 ARM8, but does not specifically bind human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, murine proMyostatin, human latent Myostatin, human GDF-8 prodomain, human GDF- 11 ARM8 prodomain, and human proGDFl 1 ARM8, but does not specifically bind human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyo statin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyo statin has an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human GDF-8 prodomain has an amino acid sequence as set forth in SEQ ID NO: 38.
  • the human GDF-11 ARM8 prodomain has an amino acid sequence as set forth in SEQ ID NO: 181.
  • the human proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the Myostatin binding profile further comprises that the antibody specifically binds human latent proGDFl 1 ARM8, but does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4. In some embodiments, the Myostatin binding profile further comprises that the antibody specifically binds murine latent myostatin and human latent proGDFl 1 ARM8, but does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4. In some embodiments, the murine latent myostatin has an amino acid sequence as set forth in SEQ ID NO: 88. In some embodiments, the human latent proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 752, 780, 815, 772, 784, 770, 774, 756, 796, 785, 761, 788, 767, 765, 799, 749, 802, 809, 750, 803, 800, 806, 759, 805, 797, 787, or 766, 782, 967, 776, 755, 876, 891, 963, 996, 993, 970, 968 or 1008.
  • the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1367, 1394, 1660, 1387, 1397, 1385, 1388, 1371, 1409, 1398, 1376, 1401, 1382, 1380, 1656, 1364, 1413, 1418, 1365, 1658, 1657, 1415, 1374, 1410, 1400, 1381, 1395, 1582, 1390, 1370, 1670, 1485, 1578, 1611, 1608, 1585, 1583, or 1623.
  • the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP6a-l, BP6a-2, BP6a-3, BP6a-4, BP6a-5, BP6a-6, BP6a-7, BP6a-8, BP6a-9, BP6a-10, BP6a-l l, BP6a-12, BP6a-13, BP6a-14, BP6a-15, BP6a-16, BP6a-17, BP6a-18, BP6a-19, BP6a-20, BP6a-21, BP6a-22, BP6a-23, BP6a-24, BP6a-25, BP6a-26, BP6a-27, BP6b-l, BP6b-2, BP6b-3, BP6b- 4, BP6b-5, BP6b-6,
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP6a-l, BP6a-2, BP6a-3, BP6a-4, BP6a-5, BP6a-6, BP6a-7, BP6a-8, BP6a-9, BP6a-10, BP6a-l l, BP6a-12, BP6a-13, BP6a-14, BP6a-15, BP6a-16, BP6a-17, BP6a-18, BP6a-19, BP6a-20, BP6a-21, BP6a-22, BP6a-23, BP6a-24, BP6a-25, BP6a-26, BP6a-27, BP6b-l, BP6b-2, BP6b-3, BP6b-4, BP6b-5, BP6b-6, BP6b-7, BP6b-8, BP6b-9, BP6b-10, BP6b
  • the antibody comprises the variable light chain amino acid sequence of clone BP6a-l, BP6a-2, BP6a-3, BP6a-4, BP6a-5, BP6a-6, BP6a-7, BP6a-8, BP6a-9, BP6a- 10, BP6a-l l, BP6a-12, BP6a-13, BP6a-14, BP6a-15, BP6a-16, BP6a-17, BP6a-18, BP6a-19, BP6a-20, BP6a-21, BP6a-22, BP6a-23, BP6a-24, BP6a-25, BP6a-26, BP6a-27, BP6b-l, BP6b-2, BP6b-3, BP6b-4, BP6b-5, BP6b-6, BP6b-7, BP6b-8, BP6b-9, BP6b-10, BP6b-
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, and human proGDFl 1 ARM8 but does not specifically bind human latent Myostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, murine proMyostatin and human proGDFl 1 ARM8 but does not specifically bind human latent Myostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 813.
  • antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1422.
  • the antibody comprises a CDR- Hl, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP2a-l as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP2a-l as set forth in Table 16.
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP2a-l as set forth in Table 14.
  • the antibody comprises the variable light chain amino acid sequence of clone BP2a-l as set forth in Table 16.
  • the myostatin binding profile is that the antibody
  • human proMyostatin specifically binds human proMyostatin, and human latent Myostatin, but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179, or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, murine proMyostatin, and human latent Myostatin, but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179, or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 794, 804, or 751.
  • the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1407, 1414, or 1366.
  • the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP3a-l, BP3a-2 or BP3a-3 as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP3a-l, BP3a-2 or BP3a-3 as set forth in Table 16.
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP3a-l, BP3a-2 or BP3a-3 as set forth in Table 14.
  • the antibody comprises the variable light chain amino acid sequence of clone BP3a-l, BP3a-2 or BP3a-3 as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin and human GDF8 prodomain but does not specifically bind human GDF- 11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, murine proMyostatin, human latent Myostatin and human GDF8 prodomain but does not specifically bind human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human GDF-8 prodomain has an amino acid sequence as set forth in SEQ ID NO: 38.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 768. In some embodiments, the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1383. In some embodiments, the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP4a-l as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP4a-l as set forth in Table 16.
  • VH variable heavy chain
  • VL variable light chain
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP4a-l as set forth in Table 14. In some embodiments, the antibody comprises the variable light chain amino acid sequence of clone BP4a-l as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin, and human proGDFl 1 ARM8, but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, murine proMyostatin, human latent Myostatin, and human proGDFl 1 ARM8, but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the Myostatin binding profile further comprises that the antibody specifically binds murine latent myostatin and human latent proGDFl 1 ARM8, but does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4.
  • the Myostatin binding profile further comprises that the antibody specifically binds murine latent myostatin and human latent proGDFl 1 ARM 8, but does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4.
  • the murine latent myostatin has an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 760, 798, 792, 789, 801, 790, 812, 758, 762, 775, 834, 974, 985, or 1002 .
  • the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1375, 1411, 1405, 1402, 1412, 1403, 1421, 1373, 1377, 1389, 1661, 1589, 1600, or 1617 .
  • the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP5a-l, BP5a-2, BP5a-3, BP5a-4 or BP5a-5, BP5b-l, BP5b-2, BP5b-3, BP5b-4, BP5b-5, BP5b-6, BP5b-7, BP5b-8 or BP5b-9 as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP5a-l, BP5a-2, BP5a-3, BP5a-4 or BP5a-5, BP5b-l, BP5b-2, BP5b-3, BP5b-4, BP5b-5, BP5b-6, BP5
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP5a-l, BP5a-2, BP5a-3, BP5a-4 or BP5a-5, BP5b-l, BP5b-2, BP5b- 3, BP5b-4, BP5b-5, BP5b-6, BP5b-7, BP5b-8 or BP5b-9 as set forth in Table 14.
  • the antibody comprises the variable light chain amino acid sequence of clone BP5a-l, BP5a-2, BP5a-3, BP5a-4 or BP5a-5, BP5b-l, BP5b-2, BP5b-3, BP5b-4, BP5b-5, BP5b-6, BP5b-7, BP5b-8 or BP5b-9 as set forth in Table 16.
  • the myostatin binding profile is that the antibody
  • human proMyostatin specifically binds human proMyostatin but does not specifically bind human latent Myostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179, or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyo statin and murine proMyostatin but does not specifically bind human latent Myostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179, or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 88).
  • the Myostatin binding profile further comprises that the antibody does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4 or human latent proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179.
  • the Myostatin binding profile further comprises that the antibody does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4, murine latent myostatin having an amino acid sequence as set forth in SEQ ID NO: 88 or human latent proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence as set forth in SEQ ID NO: 769 or 827.
  • the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1384 or 1435.
  • the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BPla-1 or BPlb-1 as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BPla-1 or BPlb-1 as set forth in Table 16.
  • the antibody comprises a variable heavy chain amino acid sequence of clone BPla-1 or BPlb-1 as set forth in Table 14.
  • the antibody comprises the variable light chain amino acid sequence of clone BPla-1 or BPlb-1 as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human latent Myostatin but does not specifically bind human proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179, human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42 or human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4).
  • the myostatin binding profile is that the antibody specifically binds human latent Myostatin but does not specifically bind murine proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 88, human proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179, human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42 or human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4).
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the Myostatin binding profile further comprises that the antibody specifically binds human latent proGDFl 1 ARM8.
  • the Myostatin binding profile further comprises that the antibody specifically binds murine latent myostatin and human latent proGDFl 1 ARM8.
  • the murine latent myostatin has an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO990.
  • the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1605. In some embodiments, the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BPlOb-1 as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BPlOb-1 as set forth in Table 16. In some embodiments, the antibody comprises a variable heavy chain amino acid sequence of clone BPlOb-1 as set forth in Table 14. In some embodiments, the antibody comprises the variable light chain amino acid sequence of clone BP 10b- las set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin, human proGDFl 1 ARM8 and human mature GDF-8 but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38 or human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, murine proMyostatin, human latent Myostatin, human proGDFl 1 ARM8 and human mature GDF-8 but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38 or human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the human mature GDF-8 has an amino acid sequence as set forth in SEQ ID NO: 42.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 764. In some embodiments, the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1679.
  • the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR- H3 from the variable heavy chain (VH) amino acid sequence of clone BP1 la-1 as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP1 la-1 as set forth in Table 16.
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP1 la-1 as set forth in Table 14.
  • the antibody comprises the variable light chain amino acid sequence of clone BP 1 la-1 as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human latent Myostatin and human mature GDF-8 but does not specifically bind human proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF- 11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181 or human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179.
  • the myostatin binding profile is that the antibody specifically binds human latent Myostatin and human mature GDF-8 but does not specifically bind murine proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 88, human proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181 or human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179.
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human mature GDF-8 has an amino acid sequence as set forth in SEQ ID NO: 42.
  • the Myostatin binding profile further comprises that the antibody does not specifically bind human proGDFl l having an amino acid sequence as set forth in SEQ ID NO: 4 or human latent proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179.
  • the Myostatin binding profile further comprises that the antibody specifically binds murine latent myostatin but does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4 or human latent proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179.
  • the murine latent myostatin has an amino acid sequence as set forth in SEQ ID NO: 88.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 989.
  • the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1604.
  • the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP12b-l as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP12b-las set forth in Table 16.
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP12b-l as set forth in Table 14.
  • the antibody comprises the variable light chain amino acid sequence of clone BP12b-l as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin and human GDF-8 prodomain, but does not specifically bind human latent Myostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin and human GDF-8 prodomain, but does not specifically bind murine proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 88, human latent Myostatin having an amino acid sequence as set forth in SEQ ID NO: 5, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human GDF-8 prodomain has an amino acid sequence as set forth in SEQ ID NO: 38.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 786.
  • the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1399.
  • the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR- H3 from the variable heavy chain (VH) amino acid sequence of clone BP15a-l as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP15a-l as set forth in Table 16.
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP15a-l as set forth in Table 14.
  • the antibody comprises the variable light chain amino acid sequence of clone BP 15a- 1 as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin and human latent Myostatin but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin and human latent Myostatin but does not specifically bind murine proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 88, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the Myostatin binding profile further comprises that the antibody does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4 or human latent proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179.
  • the Myostatin binding profile further comprises that the antibody does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4, murine latent myostatin having an amino acid sequence as set forth in SEQ ID NO: 88 or human latent proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 810, 855, 971, 983, or 986.
  • the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1419, 1662, 1586, 1598, or 1601.
  • the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP16a-l, BP16b-l, BP16b-2, BP16b-3 or BP16b-4 as set forth in Table 14, and/or a CDR-Ll, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP16a-l, BP 16b-
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP16a-l, BP16b-l, BP16b-2, BP16b-3 or BP16b-4 as set forth in Table 14. In some embodiments, the antibody comprises the variable light chain amino acid sequence of clone BP16a-l, BP16b-l, BP 16b-
  • the myostatin binding profile is that the antibody specifically binds human proMyo statin and human latent Myostatin but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179, human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42 or human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin and human latent Myostatin but does not specifically bind murine proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 88, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179, human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42, human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4 or murine latent myostatin having an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human proMyostatin has amino acid sequence as set forth in SEQ ID NO: 5. In some embodiments, the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5. In some embodiments, the Myostatin binding profile further comprises that the antibody specifically binds human latent proGDFl 1 ARM8. In some embodiments, the human latent proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179. In some embodiments, the Myostatin binding profile further comprises that the antibody does not specifically bind human latent proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 982. In some embodiments, the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1597. In some embodiments, the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP17b-l as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP17b-l as set forth in Table 16.
  • VH variable heavy chain
  • VL variable light chain
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP17b-l as set forth in Table 14. In some embodiments, the antibody comprises the variable light chain amino acid sequence of clone BP17b-l as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin and human proGDFl 1 ARM8 but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin and human proGDFl 1 ARM8 but does not specifically bind murine proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 88, human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the Myostatin binding profile further comprises that the antibody specifically binds human latent proGDFl 1 ARM8 but does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4.
  • the Myostatin binding profile further comprises that the antibody specifically binds human latent proGDFl 1 ARM8 but does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4 or murine latent myostatin having an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 763, 814, 753, 777, 795, 969, 972, 976, or 981.
  • the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1378, 1423, 1368, 1391, 1408, 1584, 1587, 1591, or 1596.
  • the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP18a-l or BP18a-2, BP18b-l, BP18b-2, BP18b-3, BP18b-4, BP18b-5, BP18b-6 or BP18b-7 as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP18a-l or BP18a-2, BP18b-l, BP18b-2, BP18b-3, BP18b-4, BP18b-5, BP18b-6 or BP18b-7as set forth in Table 16.
  • VH variable heavy chain
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP18a-l or BP18a-2, BP18b-l, BP 18b- 2, BP18b-3, BP18b-4, BP18b-5, BP18b-6 or BP18b-7 as set forth in Table 14.
  • the antibody comprises the variable light chain amino acid sequence of clone BP18a-l or BP18a-2, BP18b-l, BP18b-2, BP18b-3, BP18b-4, BP18b-5, BP18b-6 or BP18b-7 as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin, human GDF-8 prodomain, human GDF-11 ARM8 prodomain and human proGDFl 1 ARM8 but does not specifically bind human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin, human GDF-8 prodomain, human GDF-11 ARM8 prodomain and human proGDFl 1 ARM8 but does not specifically bind murine proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 88 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the antibody specifically binds human proMyostatin, human latent Myostatin, human GDF-8 prodomain, human GDF-11 ARM8 prodomain and human proGDFl 1 ARM8 but does not specifically bind murine proMyostatin having an amino acid sequence as set forth in SEQ ID NO: 88 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human GDF-8 prodomain has an amino acid sequence as set forth in SEQ ID NO: 38.
  • the human GDF-11 ARM8 prodomain has an amino acid sequence as set forth in SEQ ID NO: 181.
  • the human proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the Myostatin binding profile further comprises that the antibody specifically binds human latent proGDFl 1 ARM8 but does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4. In some embodiments, the Myostatin binding profile further comprises that the antibody specifically binds human latent proGDFl 1 ARM8 but does not specifically bind human proGDFl 1 having an amino acid sequence as set forth in SEQ ID NO: 4 or murine latent myostatin having an amino acid sequence as set forth in SEQ ID NO: 88. In some embodiments, the human latent proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 773, 807, 811, 754, 771, or 978. In some embodiments, the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1655, 1416, 1420, 1369, 1386, or 1593.
  • the antibody comprises a CDR-Hl, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP19a-l, BP19a-2 or BP19a-3, BP19b-l, BP19b-2 or BP19b-3 as set forth in Table 14, and/or a CDR- Ll, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP19a-l, BP19a-2 or BP19a-3, BP19b-l, BP19b-2 or BP19b-3 as set forth in Table 16.
  • VH variable heavy chain
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP19a-l, BP19a-2 or BP19a-3, BP19b-l, BP19b-2 or BP19b-3 as set forth in Table 14. In some embodiments, the antibody comprises the variable light chain amino acid sequence of clone BP19a-l, BP19a-2 or BP19a-3, BP19b-l, BP19b-2 or BP19b-3 as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin, human proGDFl 1 ARM8, human proGDFl 1 or human latent proGDFl 1 ARM8 but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, murine proMyostatin, human latent Myostatin, human proGDFl 1 ARM8, human proGDFl 1, murine latent myostatin or human latent proGDFl 1 ARM8 but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 88. In some embodiments, the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5. In some embodiments, the human proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179. In some embodiments, the human proGDFl 1 has an amino acid sequence as set forth in SEQ ID NO: 4. In some embodiments, the murine latent myostatin has an amino acid sequence as set forth in SEQ ID NO: 88. In some embodiments, the human latent proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 779. In some embodiments, the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1393. In some embodiments, the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP21b-l as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP21b-l as set forth in Table 16.
  • VH variable heavy chain
  • VL variable light chain
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP21b-l as set forth in Table 14. In some embodiments, the antibody comprises the variable light chain amino acid sequence of clone BP21b-l as set forth in Table 16.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, human latent Myostatin, human proGDFl 1 and human latent proGDFl 1 ARM8 but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the myostatin binding profile is that the antibody specifically binds human proMyostatin, murine proMyostatin, human latent Myostatin, human proGDFl 1, murine latent myostatin and human latent proGDFl 1 ARM8 but does not specifically bind human GDF-8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 38, human GDF-11 ARM8 prodomain having an amino acid sequence as set forth in SEQ ID NO: 181, human proGDFl 1 ARM8 having an amino acid sequence as set forth in SEQ ID NO: 179 or human mature GDF-8 having an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 5. In some embodiments, the murine proMyostatin has an amino acid sequence as set forth in SEQ ID NO: 88. In some embodiments, the human latent Myostatin has an amino acid sequence as set forth in SEQ ID NO: 5. In some embodiments, the human proGDFl 1 has an amino acid sequence as set forth in SEQ ID NO: 4. In some embodiments, the murine latent myostatin has an amino acid sequence as set forth in SEQ ID NO: 88. In some embodiments, the human latent proGDFl 1 ARM8 has an amino acid sequence as set forth in SEQ ID NO: 179.
  • the antibody comprises a CDR-H3 amino acid sequence set forth in SEQ ID NO: 1006. In some embodiments, the antibody comprises a CDR-L3 amino acid sequence set forth in any one of SEQ ID NO: 1621. In some embodiments, the antibody comprises a CDR-H1, a CDR-H2, and/or a CDR-H3 from the variable heavy chain (VH) amino acid sequence of clone BP22b-l as set forth in Table 14, and/or a CDR-L1, a CDR-L2, and/or a CDR-L3 from the variable light chain (VL) amino acid sequence of clone BP22b-las set forth in Table 16.
  • VH variable heavy chain
  • VL variable light chain
  • the antibody comprises a variable heavy chain amino acid sequence of clone BP22b-l as set forth in Table 14. In some embodiments, the antibody comprises the variable light chain amino acid sequence of clone BP22b-l as set forth in Table 16.
  • the biological sample is subjected to an immunoassay using two or more antibodies, and wherein at least two of the antibodies have different myostatin binding profiles.
  • the biological sample is subjected to an immunoassay using two or more antibodies, and wherein at least two of the antibodies have different myostatin binding profiles.
  • the biological sample is subjected to an immunoassay using two or more antibodies, and wherein at least two of the antibodies have different myostatin binding profiles.
  • the biological sample is subjected to an immunoassay using two or more antibodies, and wherein at least two of the antibodies have different myostatin binding profiles.
  • the biological sample is subjected to an immunoassay using four or more antibodies, and wherein at least four of the antibodies have different myostatin binding profiles.
  • the biological sample is from a subject.
  • the biological sample is a blood sample.
  • the biological sample is a tissue sample.
  • the tissue sample is from muscle, heart, spleen, kidney, liver, or adipose tissue.
  • the tissue sample is from skeletal muscle or cardiac muscle.
  • the tissue sample is from a muscle biopsy.
  • the immunoassay is an enzyme-linked
  • an immunohistochemical assay that provides information regarding spatial location and/or level of different myostatin forms based on the selected Myostatin binding profile.
  • the selected Myostatin binding profile In some embodiments, the
  • the tissue sample is a formalin-fixed paraffin-embedded tissue (FFPE) sample.
  • the tissue sample is cryo-preserved.
  • the tissue sample is from a subject.
  • the method further includes determining the spatial location and/or level of the different myostatin forms in the tissue sample.
  • the method further includes evaluating the subject based on the spatial location and/or level of the different myostatin forms in the tissue sample.
  • the method further includes treating the subject based on the spatial location and/or level of the different myostatin forms in the tissue sample.
  • the subject has or is suspected of having TGF-associated condition.
  • the TGF-associated condition comprises muscle atrophy.
  • the TGF-associated condition is muscular dystrophy, cachexia, sarcopenia or atrophy associated with disuse.
  • the immunoassay is an immunoprecipitation assay.
  • the immunoprecipitation assay that provides information regarding the level and/or ratio of different myostatin forms based on the selected Myostatin binding profile.
  • the immunoprecipitation assay is performed using a fluid sample.
  • the fluid sample comprises whole blood, plasma, serum, exosome-like vesicles from blood, saliva, cerebrospinal fluid, or urine.
  • the fluid sample comprises a lysate of a tissue.
  • the tissue is muscle, heart, spleen, kidney, liver, or adipose tissue.
  • the tissue sample is skeletal muscle or cardiac muscle. In some embodiments, the sample is from a subject. In some embodiments, the level and/or ratio of the different myostatin forms in the sample. In some embodiments, the method further comprises diagnosing the subject based on the level and/or ratio of the different myostatin forms in the sample. In some embodiments, the method further comprises treating the subject based on the level and/or the ratio of the different myostatin forms in the tissue sample. In some embodiments, the subject has or is suspected of having TGF-associated condition. In some embodiments, the TGF-associated condition comprises muscle atrophy. In some embodiments, the TGF-associated condition is muscular dystrophy, cachexia, sarcopenia or atrophy associated with disuse.
  • the immunoassay is an enzyme linked immunosorbent assay (ELISA).
  • the ELISA provides information regarding the level and/or ratio of different myostatin forms based on the selected Myostatin binding profile.
  • the ELISA is performed using a fluid sample.
  • the fluid sample comprises whole blood, plasma, serum, exosome-like vesicles from blood, saliva, cerebrospinal fluid, or urine.
  • the fluid sample comprises a lysate of a tissue.
  • the tissue is muscle, heart, spleen, kidney, liver, or adipose tissue.
  • the tissue sample is skeletal muscle or cardiac muscle.
  • the sample is from a subject. In some embodiments, the method further comprises determining the level and/or ratio of the different myostatin forms in the sample. In some embodiments, the method further comprises diagnosing the subject based on the level and/or ratio of the different myostatin forms in the sample. In some
  • the method further comprises treating the subject based on the level and/or the ratio of the different myostatin forms in the tissue sample.
  • the subject has or is suspected of having TGF-associated condition.
  • the TGF- associated condition comprises muscle atrophy.
  • the TGF-associated condition is muscular dystrophy, cachexia, sarcopenia or atrophy associated with disuse.
  • the immunoassay is a Western blot assay.
  • the Western blot assay is a quantitate Western blot assay.
  • the Western blot assay is a capillary Western blot assay.
  • the Western blot provides information regarding the level and/or ratio of different myostatin forms based on the selected Myostatin binding profile.
  • the Western blot is performed using a fluid sample.
  • the fluid sample comprises whole blood, plasma, serum, exosome-like vesicles from blood, saliva, cerebrospinal fluid, or urine.
  • the fluid sample comprises a lysate of a tissue.
  • the tissue is muscle, heart, spleen, kidney, liver, or adipose tissue.
  • the tissue sample is skeletal muscle or cardiac muscle.
  • the sample is from a subject.
  • the method of further comprises determining the level and/or ratio of the different myostatin forms in the sample. In some embodiments, the method further comprises diagnosing the subject based on the level and/or ratio of the different myostatin forms in the sample. In some
  • the method further comprises treating the subject based on the level and/or the ratio of the different myostatin forms in the tissue sample.
  • the subject has or is suspected of having TGF-associated condition.
  • the TGF- associated condition comprises muscle atrophy.
  • the TGF-associated condition is muscular dystrophy, cachexia, sarcopenia or atrophy associated with disuse.
  • FIG. 1 is a diagram of the TGF-beta superfamily tree, where divergence is proportional to branch length.
  • FIG. 2 is a schematic of one embodiment of a linear representation of a translated growth factor monomer.
  • translated growth factors may comprise secretion signal peptides, prodomains and growth factor domains.
  • translated growth factors may also comprise a cleavage site between prodomain and growth factor regions.
  • FIG. 3 is a schematic of one embodiment of a GDF growth factor-prodomain complex (GPC) made up of proGDF monomers as well as an embodiment of a free growth factor dimer and free prodomains, separated after proteolytic cleavage of the GPC.
  • GPC GDF growth factor-prodomain complex
  • FIG. 4 is a schematic of one embodiment of a GDF GPC as well as an embodiment of a free growth factor dimer and free prodomains, separated after proteolytic cleavage.
  • FIG. 5 presents 3 tables showing the percent identity between amino acid sequences found in the TGF- ⁇ family.
  • FIG. 5A demonstrates percent identity among pro- proteins (prodomain and growth factor). Percent identity among growth factor domains is presented in FIG. 5B while percent identity among prodomains is presented in FIG. 5C.
  • FIG. 6 shows structure-based alignment between TGF- ⁇ family member proteins [adapted from Shi et al (Shi, M. et al., Latent TGF- ⁇ structure and activation. Nature. 2011 Jun 15; 474(7351):343-9, the contents of which are herein incorporated by reference in their entirety)] .
  • Cysteine residues required for interaction with LTBPs and/or GARPs are boxed.
  • Residues mutated in Camurati-Engelmann syndrome are indicated with a star.
  • Protease cleavage sites are indicated with an up arrow.
  • Protein modules and secondary structural elements are indicated with solid bars.
  • Residues underlined at the N-terminus of GDF-8 correspond to alternatively predicted signal peptide processing sites.
  • “Chimeric module breakpoints” indicate regions where structural features are conserved and provide modules for chimeric protein construction (swapping of modules between family members) in all family members. N-terminal regions are shown in (A), internal regions are shown in (B) and C-terminal regions are shown in (C).
  • FIG. 7 is a schematic of an embodiment of a recombinant GPC.
  • FIG. 8 is a schematic of embodiments of mutant recombinant proteins of the disclosure.
  • FIG. 9 depicts the formation of a latent complex after furin cleavage of a pro- protein dimer. The latent complex retains the growth factor dimer until subjected to an activating cleavage [e.g. by members of the BMP- 1/Tolloid- like proteinase (B/TP) family.]
  • B/TP BMP- 1/Tolloid- like proteinase
  • FIG. 10 presents an alignment conducted between GDF-8 (myostatin), GDF-11, Inhibin A and a GDF-8 dimer. Arrows indicate cleavage sites. Regions involved in internal interactions are boxed. Solid rectangles appear above residues predicted to be involved in steric clashes in chimeric constructs. Stars denote important break points in protein modules.
  • FIG. 11 depicts recombinant histidine-tagged proGDF-8 (referred to in the figure as proMyostatin), separated by SDS-PAGE under reducing and non-reducing conditions, as visualized by colloidal blue staining.
  • proMyostatin histidine-tagged proGDF-8
  • FIG. 12 is a graph showing CAGA promoter-dependent luciferase activity is the presence of GDF-11 or proGDF-11 after treatment with proprotein convertase, Tolloid proteinase or a combination of proprotein convertase and Tolloid proteinase.
  • FIG. 13 is a stained gel showing separation of proteinase treated proGDF-11 under reducing and non-reducing conditions.
  • FIG. 14 presents results of a luciferase-based growth factor activity assay.
  • FIG. 15 is a schematic showing formation of the "primed" complex.
  • the primed complex is an open, active conformation of GDF-8 GPCs or GDF-11 GPCs. Primed complexes are capable of receptor binding and inducing growth factor activity.
  • FIG. 16 is a graph showing that BP6a-5 dissociates rapidly at pH 5. The antibody blocks myostatin activation and promotes muscle growth in vivo.
  • FIG. 17 is a graph showing that BP6a-6 dissociates rapidly at pH 5.
  • the antibody does not block Myostatin activation or signaling and is in the same epitope bin (by ELISA binding profile) as BP6a-5.
  • FIG. 18 displays eight antibodies that dissociate more rapidly at pH5 that at pH 7.4.
  • the asterisk indicates some cross-reactivity with proGDFl 1 in ELISA and the bold indicates that the antibody inhibits GDF8 activation.
  • FIG. 19 shows CDR-H3 sequences of pH-sensitive antibodies. All listed antibodies have a histidine in the CDR-H3 sequence except for BP5b- 1. The sequences, from top to bottom, correspond to SEQ ID NOs: 762, 764, 770, 777, 784, 986, 1006, and 790. Note that BP18b-2 only includes one histidine in the entire variable region, while BP5b-l does not contain any histidines in the variable region. These two antibodies have the weakest pH sensitivity profiles. [0048] FIG. 20 is a table of four antibodies for in vivo characterization of Myostatin sweeping. All antibodies will be produced both with and without the "sweeping" mutations in the constant region. Light grey indicates the antibody inhibits GDF8 activation.
  • FIG. 21 is a graph showing that BP1 la-1 dissociates rapidly at pH5.
  • the antibody does not block Myostatin activation or signaling, but does bind to proMyostatin, latent myostatin, and mature myostatin with equal affinity.
  • FIG. 22 is a graph showing that BP5b-4 dissociates rapidly at pH5.
  • the antibody does not block Myostatin activation or signaling; however, it is a different epitope group from BP1 la-1 and BP6a-5.
  • FIG. 23 shows an example of an antibody (BP6a-3) with no pH-sensitive binding.
  • FIGs. 24A-24L show the different pH sensitivities of specific antibodies: BP6a-3 (FIG. 24A), BP5a-l (FIG. 24B), BP6a-l l (FIG. 24C), BP5b-4 (FIG. 24D), BPl la-1 (FIG. 24E), BP6a-6 (FIG. 24F), BP18b-2 (FIG. 24G), BP6a-2 (FIG. 24H), BP6a-5 (FIG. 241), BP14a-l (FIG. 24J), BP5b-2 (FIG. 24K), and BP18b-7 (FIG. 24L).
  • FIGs. 25A-25K show the different pH sensitivities of specific antibodies: BP16b-4 (FIG. 25 A), BP6b-9 (FIG. 25B), BP22b-l (FIG. 25C), BP3a-2 (FIG. 25D), BP5b-l (FIG. 25E), BP6a-4 (FIG. 25F), BP6a-7 (FIG. 25G), BPla-1 (FIG. 25H), BP2a-l (FIG. 251), BP5a- 2 (FIG. 25J), and BP6a-l (FIG. 25K).
  • antibodies are selected for use (e.g., in a particular assay) based on the fact that they have a known and/or desired binding profile (a selected binding profile).
  • binding profile refers to a set of one or more parameters (e.g., symbols, quantities, measurements, etc.) indicative of the extent to which an antibody specifically binds to one or more antigens.
  • a parameter indicative of the binding of an antibody for a target antigen is an IC50 or EC50 value.
  • a parameter indicative of the binding of an antibody for a target antigen is an equilibrium dissociation constant (Kd).
  • a parameter indicative of the binding is an equilibrium association constant (Ka).
  • Ka equilibrium association constant
  • the disclosure relates to antibodies having a selected myo statin-related binding profile, which comprises a set of one or more parameters indicative of the extent to which an antibody specifically binds to one or more antigens, at least one of which antigens is a myostatin-related protein (e.g., Myostatin, GDF11, etc.).
  • a binding profile comprises one or more parameters indicative of whether or not an antibody exhibits a threshold level of binding (e.g., specific binding) to one or more antigens.
  • a threshold level of binding is a level of binding that above (or below, depending on the parameter) a control or reference level of binding (e.g., background or non-specific binding).
  • a threshold level of binding is a level of binding that is at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7 or more standard deviations greater than (or less than, depending on the assay and/or parameter) a control or reference level of binding (e.g., background or non-specific binding), as measured by an appropriate immunoassay.
  • a threshold level of binding is a level of binding that is in a range of 1 to 10 standard deviations, 2 to 10 standard deviations , or 4 to 6 standard deviations greater than (or less than, depending on the assay and/or parameter) a control or reference level of binding (e.g., background or non-specific binding), as measured by an appropriate immunoassay.
  • a threshold level of binding is determined through an appropriate immunoassay.
  • an appropriate immunoassay assesses the binding affinity of an antibody for a target antigen.
  • an appropriate immunoassay is an enzyme linked immune- sorbent assay.
  • an appropriate immunoassay is an assay that determines a kinetic measurement (e.g., on rate, off rate) indicative of binding between an antibody and antigen.
  • an appropriate immunoassay is an assay, such as an Octet assay, that determines one or more a kinetic parameters indicative of binding between an antibody and antigen.
  • an appropriate immunoassay is a cell-based assay that determines one or more a parameters indicative of specific binding between an antibody and antigen based on cellular activity, e.g., growth factor signaling (such as SMAD signaling), cell growth, cell survival, gene expression, reporter expression, protein production, protein secretion, etc.
  • an appropriate immunoassay is an in vivo assay that determines one or more a parameters indicative of specific binding between an antibody and antigen based on cellular, tissue or other physiological activity.
  • a binding profile comprises a set of one or more parameters indicative of the extent to which an antibody specifically binds to one or more one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins.
  • a binding profile comprises a set of one or more parameters indicative of the extent to which an antibody specifically binds or does not specifically bind to one or more different antigens.
  • a binding profile relates to the extent to which an antibody specifically binds or does not specifically bind to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more different antigens. In some embodiments, a binding profile relates to the extent to which an antibody specifically binds or does not specifically bind to 5, 6, 7, 8, 9, 10 or 11 different antigens.
  • a particular binding profile comprises a set of one or more parameters indicative of the extent to which an antibody specifically binds to one or more TGFP family member proteins or forms thereof.
  • a TGFP family member proteins is selected from the group consisting of AMH, ARTN, BMP10, BMP15, BMP2, BMP3, BMP4, BMP5, BMP6, BMP7, BMP8A, BMP8B, GDF1, GDF10, GDF11, GDF15, GDF2, GDF3, GDF3A, GDF5, GDF6, GDF7, GDF8, GDF9, GDNF, INHA, INHBA, INHBB, INHBC, INHBE, LEFTY 1, LEFTY2, NODAL, NRTN, PSPN, TGFpi, TGFP2, and TGFP3 protein.
  • TGFP family member proteins or forms thereof are from a vertebrate organism.
  • TGFP family member proteins or forms thereof are from a vertebrate organism.
  • TGFP family member proteins or forms thereof
  • TGFP family member proteins or forms thereof are from a human or a mouse. In some embodiments, TGFP family member proteins or forms thereof are from a human. Examples of sequences of human and non-human TGFP family member proteins are shown in Tables 1, 4 and 5, provided herein. In some embodiments, TGFP family member proteins or forms thereof may include any naturally- occurring isoforms or variants of TGFP family member proteins. In some embodiments, TGFP family member proteins comprise an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to any of the amino acid sequences as set forth in any one of SEQ ID NOs: 1-37 and 70-115.
  • TGFP family member proteins comprise an amino acid sequence as set forth in any one of SEQ ID NOs: 1-37 and 70-115. In some embodiments, TGFP family member proteins consist of an amino acid sequence as set forth in any one of SEQ ID NOs: 1-37 and 70-115.
  • a particular binding profile may also comprise one or more parameters indicative of to the extent to which antibodies specifically bind to or do not specifically bind to a portion or domain of one or more TGFP family member proteins.
  • the portion or domain of a TGFP family member protein is a prodomain, a straight jacket region, a growth factor domain, a fastener region, a furin cleavage site region, a bmp/tolloid cleavage site, an arm region, a fingers region 1, a fingers region 2, a latency loop, an alpha 1 helical region, and/or a bowtie region.
  • Exemplary portions or domains of TGFP family member proteins are shown in Tables 2, 3, 9 and 11, provided herein.
  • portions or domains of TGFP family member proteins are portions or domains of GDF proteins. In some embodiments, portions or domains of TGFP family member proteins are portions or domains of GDF8 and/or GDF11. In some embodiments, portions or domains of TGFP family member proteins are portions or domains of Inhibin beta A. However, it should be appreciated that the portions or domains of TGFP family member proteins may be from any TGFP family member protein provided herein.
  • the portion or domain of a TGFP family member protein comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to any of the amino acid sequences as set forth in any one of SEQ ID NOs: 38-65, 162-177 and 202-221.
  • portions or domains of TGFP family member proteins comprise an amino acid sequence as set forth in any one of SEQ ID NOs: 38-65, 162-177 and 202-221.
  • portions or domains of TGFP family member proteins consist of an amino acid sequence as set forth in any one of SEQ ID NOs: 38-65, 162- 177 and 202-221.
  • forms of TGFP family member proteins refer to complexes of TGFP family member proteins.
  • forms of TGFP family member proteins may include pro-forms, latent-forms, primed-forms, or mature forms of dimeric TGFP family member proteins, such as, for example, proMyostatin, proGDFl 1, latent Myostatin, latent GDF11, primed Myostatin, and primed GDF11.
  • TGFP family member proteins form dimeric complexes.
  • TGFP family member proteins form homodimeric complexes.
  • TGFP family member proteins form heterodimeric complexes.
  • Dimeric forms of TGFP family member proteins may include TGFP family member proteins that are full-length or TGFP family member proteins that have been cleaved (e.g., by a proprotein convertase and/or a tolloid protease).
  • forms of TGFP family member proteins are pro forms of TGFP family member proteins (e.g., proMyostatin or proGDFl 1).
  • forms of TGFP family member proteins include full-length TGFP family member proteins.
  • pro forms of TGFP family member proteins include, without limitation, proMyostatin and proGDFl 1 that have not been cleaved at a proprotein convertase cleavage site (e.g., by a proprotein convertase such as furin) or a tolloid protease cleavage site (e.g., by a tolloid protease such as BMP-1).
  • forms of TGFP family member proteins are latent forms of TGFP family member proteins (e.g., latent Myostatin or latent GDFl 1).
  • forms of TGFP family member proteins include TGFP family member proteins that have been cleaved (e.g., by a proprotein convertase).
  • latent forms of TGFP family member proteins include, without limitation, latent Myostatin and latent GDFl 1 that have been cleaved at a proprotein convertase cleavage site (e.g., by a proprotein convertase such as furin) but not at a tolloid protease cleavage site.
  • forms of TGFP family member proteins are primed forms of TGFP family member proteins (e.g., primed Myostatin or primed GDFl 1).
  • forms of TGFP family member proteins include TGFP family member proteins that have been cleaved (e.g., by a proprotein convertase and/or a tolloid protease).
  • primed forms of TGFP family member proteins include, without limitation, primed Myostatin and primed GDFl 1 that have been cleaved at a proprotein convertase cleavage site (e.g., by a proprotein convertase such as furin) and a tolloid protease cleavage site (e.g., by a tolloid protease such as BMP-1).
  • forms of TGFP family member proteins are mature forms of TGFP family member proteins (e.g., mature Myostatin or mature GDFl 1).
  • forms of TGFP family member proteins include TGFP family member proteins that have been cleaved (e.g., by a proprotein convertase and/or a tolloid protease) and are not in complex with one or more portions of a prodomain of a TGFP family member protein.
  • mature forms of TGFP family member proteins include, without limitation, mature Myostatin and mature GDFl 1 that have been cleaved at a proprotein convertase cleavage site (e.g., by a proprotein convertase such as furin), a tolloid protease cleavage site (e.g., by a tolloid protease such as BMP-1), and are not in complex with a prodomain of a TGFP family member protein.
  • a proprotein convertase cleavage site e.g., by a proprotein convertase such as furin
  • a tolloid protease cleavage site e.g., by a tolloid protease such as BMP-1
  • a particular binding profile comprises a set of one or more parameters indicative of the extent to which antibodies specifically bind to or do not specifically bind to chimeras of TGFP family member proteins.
  • chimeras of TGFP family member proteins can be used to provide information relating to particular epitopes to which any of the antibodies provided herein specifically bind or do not specifically bind.
  • chimeric TGFP family member proteins comprise at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 portions or domains of different TGFp family member protein.
  • a chimeric TGFP family member protein may comprise a prodomain, a straight jacket region, a growth factor domain, a fastener region, a furin cleavage site region, a bmp/tolloid cleavage site, an arm region, a fingers region 1, a fingers region 2, a latency loop, an alpha 1 helical region, and/or a bowtie region of one TGFP family member protein and at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 of a prodomain, a straight jacket region, a growth factor domain, a fastener region, a furin cleavage site region, a bmp/tolloid cleavage site, an arm region, a fingers region 1, a fingers region 2, a latency loop, an alpha 1 helical region, and/or a bowtie region from one or more different TGFP family member proteins.
  • chimeras of TGFP family member proteins are shown in Table 10, provided herein.
  • chimeras of TGFP family member proteins comprise an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to any of the amino acid sequences as set forth in any one of SEQ ID NOs: 178-201.
  • chimeras of TGFP family member proteins comprise an amino acid sequence as set forth in any one of SEQ ID NOs: 178-201.
  • chimeras of TGFP family member proteins consist of an amino acid sequence as set forth in any one of SEQ ID NOs: 178-201.
  • TGFP family member proteins include Fc fusion proteins to any of the TGFP family member proteins or forms or domains or portions thereof.
  • TGFP family member Fc fusion proteins are shown in Table 12, provided herein.
  • TGFP family member Fc fusion proteins comprise an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to any of the amino acid sequences as set forth in any one of SEQ ID NOs: 223-226.
  • TGFP family member Fc fusion proteins comprise an amino acid sequence as set forth in any one of SEQ ID NOs: 223-226.
  • TGFP family member Fc fusion proteins consist of an amino acid sequence as set forth in any one of SEQ ID NOs: 223-226.
  • a particular binding profile comprises a set of one or more parameters indicative of the extent to which antibodies specifically bind to or do not specifically bind to one or more of a human proMyo statin, a murine proMyostatin, a human latent Myostatin, a human GDF-8 prodomain, a human GDF-11 ARM8 prodomain, a human proGDFl 1 ARM8, a human mature GDF-8, a human proGDFl 1, a murine latent myostatin and/or a human latent proGDFl 1 ARM8.
  • parameters indicative of the extent of binding to one or more murine antigens can be removed from a binding profile.
  • a particular binding profile comprises a set of one or more parameters indicative of the extent to which antibodies specifically bind to or do not specifically bind to one or more of a human proMyostatin, a human latent
  • Myostatin a human GDF-8 prodomain, a human GDF-11 ARM8 prodomain, a human proGDFl 1 ARM8, a human mature GDF-8, a human proGDFl 1, and/or a human latent proGDFl 1 ARM8.
  • a particular binding profile relates to an extent to which antibodies specifically bind to or do not specifically bind to one or more of a protein that comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99% identical to a human proMyostatin, a murine proMyostatin, a human latent Myostatin, a human GDF-8 prodomain, a human GDF- 11 ARM8 prodomain, a human proGDFl 1 ARM8, a human mature GDF-8, a human proGDFl 1, a murine latent myostatin and/or a human latent proGDFl 1 ARM8.
  • the human proMyostatin comprises an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyostatin comprises an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent Myostatin comprises an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human GDF-8 prodomain comprises an amino acid sequence as set forth in SEQ ID NO: 38.
  • the human GDF- 11 ARM8 prodomain comprises an amino acid sequence as set forth in SEQ ID NO: 181.
  • the human proGDFl 1 ARM8 comprises an amino acid sequence as set forth in SEQ ID NO: 179.
  • the human mature GDF-8 comprises an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proGDFl 1 comprises an amino acid sequence as set forth in SEQ ID NO: 4.
  • the murine latent myostatin comprises an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent proGDFl 1 ARM8 comprises an amino acid sequence as set forth in SEQ ID NO: 179.
  • antibodies provided herein have a binding profile as set forth in Table 33.
  • a binding profile may comprises one or more symbols (e.g., +, -, +/-) indicative of the extent to which an antibody binds to an antigen.
  • binding of an antibody to an antigen at a level detectable beyond a threshold level may be indicated by a "+”.
  • a "-" indicates that the antibody does not bind the antigen at level detectable beyond a threshold in a particular assay (e.g. , is less than 2, 3, 4 or 5 standard deviations beyond a reference level, e.g. , an assay background level).
  • a "+/-" indicates that an antibody is at or near a threshold of binding the antigen as determined by a particular assay (e.g. , within 2 to 5, 3 to 5, or 4 to 5 standard deviations of a reference level).
  • antibodies that "specifically binds" to a target antigen binds to the target antigen with greater affinity, avidity, more readily, and/or with greater duration than it binds to non- target antigens.
  • antibodies provided herein have particular binding profiles, e.g., based on whether they specifically bind or do not specifically bind to one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins.
  • an antibody specifically binds an antigen if binding to that antigen is detected above a background level (e.g., of a control antigen) using an in vitro binding assay (e.g., an ELISA). In some embodiments, an antibody specifically binds an antigen if binding to that antigen is detected at least one, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 standard deviations above a background level (e.g., of a control antigen) using an in vitro binding assay.
  • an antibody specifically binds an antigen if binding to that antigen is detected at least one, at least 5 standard deviations above a background level (e.g., of a control antigen) using an in vitro binding assay.
  • the in vitro binding assay is an enzyme linked immunosorbent assay (ELISA).
  • ELISA enzyme linked immunosorbent assay
  • the ELISA is performed as described in Example 29, provided herein. However, it should be appreciated that additional methods for determining the binding affinity of a protein to an antigen are also within the scope of this disclosure
  • an antibody specifically binds to an antigen (e.g., proMyo statin) if it binds that antigen with a higher affinity as compared to another antigen (e.g., latent Myostatin). In some embodiments, an antibody specifically binds to an antigen if it binds to that antigen by at least 2-fold, 5-fold, 10-fold, 50-fold, 100-fold, 200-fold, 500- fold, or 1,000-fold higher than another antigen.
  • an antigen e.g., proMyo statin
  • an antibody specifically binds to an antigen (e.g., proMyostatin) if it binds that antigen with a higher affinity as compared to another antigen (e.g., latent Myostatin). In some embodiments, an antibody specifically binds to an antigen if it binds to that antigen with a dissociation constant (Kd) that is less than 10 "3 M, 10 "4 M, 10 "5 M, 10 "6 M, 10 "7 M, 10 "8 M
  • Kd dissociation constant
  • antibodies bind an antigen but cannot effectively eliminate the antigen from the plasma.
  • the concentration of the antigen in the plasma may be increased by reducing the clearance of the antigen.
  • antibodies e.g., sweeping antibodies provided herein have an affinity to an antigen that is sensitive to pH. Such pH sensitive antibodies may bind to the antigen in plasma at neutral pH and dissociate from the antigen in an acidic endosome, thus reducing antibody-mediated antigen accumulation and/or promoting antigen clearance from the plasma.
  • sweeping antibodies refer to antibodies having both pH-sensitive antigen binding and at least a threshold level of binding to cell surface neonatal Fc receptor (FcRn) at neutral or
  • sweeping antibodies bind to the neonatal Fc receptor FcRn at neutral pH.
  • sweeping antibodies may bind to the FcRn at a pH ranging from 7.0 to 7.6.
  • sweeping antibodies can bind to an antigen at an antigen binding site and bind to a cellular FcRn via an Fc portion of the antibody.
  • sweeping antibodies may then be internalized, releasing antigen in an acidic endosome, which may be degraded.
  • a sweeping antibody, no longer bound to the antigen may then be released (e.g., by exocytosis) by the cell back into the serum.
  • FcRn in the vascular endothelia extends the half-life of a sweeping antibody.
  • vascular endothelial cells internalize sweeping antibodies, which in some embodiments are bound to an antigen such as Myostatin (e.g., proMyostatin, latent Myostatin or primed Myostatin).
  • Myostatin e.g., proMyostatin, latent Myostatin or primed Myostatin.
  • a sweeping antibody is recycled back into the bloodstream.
  • a sweeping antibody has an increased half-life (e.g. , in the serum of a subject) as compared to its conventional counterpart.
  • a conventional counterpart of a sweeping antibody refers the antibody from which the sweeping antibody was derived (e.g., prior to engineering the Fc portion of the conventional antibody to bind FcRn with greater affinity at pH 7).
  • a sweeping antibody has a half- life in the serum of a subject that is at least 1%, 5%, 10%, 15%, 20%, 25%, 35%, 50%, 75%, 100%, 150%, 200% or 250% longer as compared to its conventional counterpart.
  • an Fc portion of a sweeping antibody binds FcRn. In some embodiments, the Fc portion of a sweeping antibody binds to FcRn at a pH of 7.4 with a Kd
  • a sweeping antibody binds to FcRn at a pH of 7.4 with a Kd ranging fromlO "3 M to 10 "7 M, from 10 "3 M to 10 "6 M, from 10 "3 M to 10 "5 M, from 10 "3 M to 10 "4 M, from 10 "4 M to 10 ⁇ 8 M, from 10 "4 M to 10 "7 M, from 10 "4 M to 10 "6 M, from 10 "4 M to 10 "5 M, from 10 "5 M to 10 ⁇ 8 M, from 10 "5 M to 10 "7 M, from 10 “5 M to 10 “6 M, from 10 “6 M to 10 “8 M, from 10 “6 M to 10 "7 M, or from 10 "7 M to 10 “8 M.
  • FcRn binds to the CH2-CH3 hinge region of a sweeping antibody. In some embodiments, FcRn binds to the same region as proteinA or protein G. In some embodiments, FcRn binds to a different binding site from FcyRs. In some embodiments, the amino acid residues AA of a sweeping antibody Fc region are required for binding to FcRn. In some embodiments, the amino acid residues AA of a sweeping antibody Fc region affect binding to FcRn.
  • any of the antibodies provided herein are engineered to bind FcRn with greater affinity. In some embodiments, any of the antibodies provided herein are engineered to bind FcRn with greater affinity at pH 7.4. In some embodiments, the affinity of sweeping antibodies to FcRn is increased to extend their pharmacokinetic (PK) properties as compared to their conventional counterparts. For example, in some embodiments, sweeping antibodies elicit less adverse reactions due to their efficacy at lower doses. In some embodiments, sweeping antibodies are administered less frequently. In some embodiments, transcytosis of sweeping antibodies to certain tissue types are increased. In some
  • sweeping antibodies enhance efficiency of trans-placental delivery. In some embodiments, sweeping antibodies are less costly to produce.
  • any of the antibodies provided herein are engineered to bind FcRn with lower affinity. In some embodiments, any of the antibodies provided herein are engineered to bind FcRn with lower affinity at pH 7.4. In some embodiments, the affinity of sweeping antibodies to FcRn is decreased to shorten their pharmacokinetic (PK) properties as compared to their conventional counterparts. For example, in some embodiments, sweeping antibodies are more rapidly cleared for imaging and/or radioimmunotherapy. In some embodiments, sweeping antibodies promote clearance of endogenous pathogenic antibodies as a treatment for autoimmune diseases. In some embodiments, sweeping antibodies reduce the risk of adverse pregnancy outcome, which may be caused by transplacental transport of material fetus- specific antibodies.
  • PK pharmacokinetic
  • sweeping antibodies have decreased affinity to an antigen at low pH as compared to a neutral or physiological pH (e.g., pH 7.4). In some embodiments, sweeping antibodies have a decreased affinity to an antigen at an acidic pH (e.g. a pH ranging from 5.5 to 6.5) as compared to a physiological pH (e.g., pH 7.4). It should be appreciated that any of the antibodies provided herein can be engineered to dissociate from the antigen depending on changes in pH (e.g., pH sensitive antibodies). In some embodiments, sweeping antibodies provided herein are engineered to bind antigen dependent on pH. In some embodiments, sweeping antibodies provided herein are engineered to bind FcRn dependent on pH.
  • sweeping antibodies provided herein are internalized by endocytosis. In some embodiments, sweeping antibodies provided here are internalized by FcRn binding. In some embodiments, endocytosed sweeping antibodies release antigen in an endosome. In some embodiments, sweeping antibodies are recycled back to the cell surface. In some embodiments, sweeping antibodies remain attached to cells. In some embodiments, endocytosed sweeping antibodies are recycled back to the plasma. It should be appreciated that the Fc portion of any of the antibodies provided herein may be engineered to have different FcRn binding activity. In some embodiments, FcRn binding activity affects the clearance time of an antigen by a sweeping antibody. In some embodiments, sweeping antibodies may be long-acting or rapid-acting sweeping antibodies.
  • converting a conventional therapeutic antibody into a sweeping antibody reduces the efficacious dose. In some embodiments, converting a conventional therapeutic antibody into a sweeping antibody reduces the efficacious dose by at least 1%, 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99%. In some embodiments, converting a conventional therapeutic antibody into a sweeping antibody reduces the efficacious dose by at least 1.5 fold, 2 fold, 3 fold, 4 fold, 5 fold, 6 fold, 8 fold, 10 fold, 15 fold, 20 fold, 50 fold or 100 fold.
  • selecting an appropriate dose of a sweeping antibody for therapy may be performed empirically.
  • a high dose of a sweeping antibody may saturate FcRn, resulting in antibodies which stabilize antigen in serum without being internalized.
  • a low dose of a sweeping antibody may not be therapeutically effective.
  • sweeping antibodies are administered once a day, once a week, once every two weeks, once every three weeks, once every four weeks, once every 6 weeks, once every 8 weeks, once every 10 weeks, once every 12 weeks, once every 16 weeks, once every 20 weeks, or once every 24 weeks.
  • any of the antibodies provided herein may be modified or engineered to be sweeping antibodies.
  • any of the antibodies provided herein may be converted into a sweeping antibody using any suitable method.
  • suitable methods for making sweeping antibodies have been previously described in Igawa et ah, (2013) "Engineered Monoclonal Antibody with Novel Antigen-Sweeping Activity In Vivo," PLoS ONE 8(5): e63236; and Igawa et al., "pH-dependent antigen-binding antibodies as a novel
  • Aspects of the disclosure relate to antibodies that compete or cross-compete with any of the antibodies provided herein.
  • the term "compete”, as used herein with regard to an antibody means that a first antibody binds to an epitope of a protein (e.g., latentMyo statin) in a manner sufficiently similar to the binding of a second antibody, such that the result of binding of the first antibody with its 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.
  • 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 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 within the scope of this disclosure. Regardless of the mechanism by which such competition or cross-competition occurs (e.g., steric hindrance, conformational change, or binding to a common epitope, or portion thereof), the skilled artisan would appreciate that such competing and/or cross-competing antibodies are encompassed and can be useful for the methods and/or compositions provided herein.
  • an antibody binds at or near the same epitope as any of the antibodies provided herein. In some embodiments, an antibody binds near an epitope if it binds within 15 or fewer amino acid residues of the epitope. In some embodiments, any of the antibodies provided herein bind within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acid residues of an epitope that is bound by any of the antibodies provided herein.
  • any of the antibodies provided herein bind at or near a tolloid cleavage site or at or near a tolloid docking site of a TGFP family member protein (e.g., proMyo statin or latent Myostatin).
  • a TGFP family member protein e.g., proMyo statin or latent Myostatin.
  • an antibody binds near a tolloid cleavage site or near a tolloid docking site if it binds within 15 or fewer amino acid residues of the tolloid cleavage site or tolloid docking site.
  • any of the antibodies provided herein bind within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acid residues of a tolloid cleavage site or tolloid docking site.
  • an antibody binds at or near a tolloid cleavage site of GDF11.
  • an antibody may bind an amino acid sequence as set forth in SEQ ID NO: 1665.
  • an antibody binds at or near a tolloid cleavage site of GDF8.
  • an antibody may bind an amino acid sequence as set forth in SEQ ID NO: 1666.
  • PKAPPLRELIDQYDVQRDDSSDGSLEDDDYHAT SEQ ID NO: 1666.
  • binding of an antibody at or near a tolloid cleavage site or at or near a tolloid docking site of a TGFP family member protein inhibits cleavage of the TGFP family member protein, for example, by a tolloid protease (e.g., BMP-1).
  • a tolloid protease e.g., BMP-1
  • any of the antibodies provided herein bind at or near a proprotein convertase cleavage site or at or near a proprotein convertase docking site of a TGFP family member protein (e.g., proMyo statin or latent Myostatin)
  • a TGFP family member protein e.g., proMyo statin or latent Myostatin
  • an antibody binds near a proprotein convertase cleavage site or near a proprotein convertase docking site if it binds within 15 or fewer amino acid residues of the proprotein convertase cleavage site or proprotein convertase docking site.
  • any of the antibodies provided herein bind within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acid residues of a proprotein convertase cleavage site or proprotein convertase docking site.
  • an antibody binds at or near a proprotein convertase cleavage site of GDF11.
  • an antibody may bind an amino acid sequence as set forth in SEQ ID NO: 1667.
  • GLHPFMELRVLENTKRSRRNLGLDCDEHSSESRC SEQ ID NO: 1667.
  • an antibody binds at or near a proprotein convertase cleavage site of GDF8.
  • an antibody may bind an amino acid sequence as set forth in SEQ ID NO: 1668.
  • GLNPFLEVKVTDTPKRSRRDFGLDCDEHSTESRC SEQ ID NO: 1668.
  • binding of an antibody at or near a proprotein convertase cleavage site or at or near a proprotein convertase docking site of a TGFP family member protein inhibits cleavage of the TGFP family member protein, for example, by a proprotein convertase (e.g., furin).
  • a proprotein convertase e.g., furin
  • an antibody competes or cross-competes for binding to any of the antigens provided herein (e.g., one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins) with an equilibrium dissociation constant, Kd, between the antibody and the protein of less than 10 "6 M.
  • an antibody competes or cross-competes for binding to any of the antigens provided herein with a Kd in a range from 10 "11 M to 10 ⁇ 6 M.
  • any of the antibodies provided herein can be characterized using any suitable methods.
  • one method is to identify the epitope to which the antigen binds, or "epitope mapping.”
  • epitope mapping There are many suitable methods for mapping and characterizing the location of epitopes on proteins, including solving the crystal structure of an antibody- antigen complex, competition assays, gene fragment expression assays, and synthetic peptide-based assays, as described, for example, in Chapter 11 of Harlow and Lane, Using Antibodies, a Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1999.
  • epitope mapping can be used to determine the sequence to which an antibody binds.
  • the epitope can be a linear epitope, i.e., contained in a single stretch of amino acids, or a conformational epitope formed by a three-dimensional interaction of amino acids that may not necessarily be contained in a single stretch (primary structure linear sequence).
  • Peptides of varying lengths e.g., at least 4-6 amino acids long
  • the epitope to which the antibody binds can be determined in a systematic screen by using overlapping peptides derived from the target antigen sequence and determining binding by the antibody.
  • the open reading frame encoding the target antigen is fragmented either randomly or by specific genetic constructions and the reactivity of the expressed fragments of the antigen with the antibody to be tested is determined.
  • the gene fragments may, for example, be produced by PCR and then
  • telomere binding domains transcribed and translated into protein in vitro, in the presence of radioactive amino acids.
  • the binding of the antibody to the radioactively labeled antigen fragments is then determined by immunoprecipitation and gel electrophoresis.
  • Certain epitopes can also be identified by using large libraries of random peptide sequences displayed on the surface of phage particles (phage libraries). Alternatively, a defined library of overlapping peptide fragments can be tested for binding to the test antibody in binding assays.
  • mutagenesis of an antigen binding domain, domain swapping experiments and alanine scanning mutagenesis can be performed to identify residues required, sufficient, and/or necessary for epitope binding.
  • domain swapping experiments can be performed using a mutant of a target antigen in which various fragments of TGFP family member proteins have been replaced (swapped) with sequences from related, but antigenically distinct proteins, such as another member of a TGFP family member protein.
  • sequences from related, but antigenically distinct proteins such as another member of a TGFP family member protein.
  • competition assays can be performed using other antibodies known to bind to the same antigen to determine whether an antibody binds to the same epitope as the other antibodies. Such competition assays would be apparent to the skilled artisan.
  • any of the suitable methods can be applied to determine whether any of the antibodies provided herein binds one or more of the specific residues/segments of one or more TGFP family member proteins as described herein. Further, the interaction of an antibody with one or more of those defined residues in TGFP family member proteins can be determined by routine technology. For example, a crystal structure can be determined, and the distances between the residues in TGFP family member proteins and one or more residues in an antibody can be determined accordingly. Based on such distance, whether a specific residue in a TGFP family member protein interacts with one or more residues in an antibody can be determined. Further, suitable methods, such as competition assays and target mutagenesis assays can be applied to determine the preferential binding of a candidate antibody to a TGFP family member protein as compared to another TGFP family member protein.
  • aspects of the disclosure relate to methods for assessing a biological sample using one or more antibodies having a selected binding profile (e.g., a binding profile as outlined in Table 33).
  • antibodies are selected for use (e.g., in an immunoassay) based on the fact that they have a known and/or desired binding profile (a selected binding profile).
  • antibodies may have a selected binding profile to one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins.
  • antibodies with selected binding profiles may be used as research and/or diagnostic tools.
  • antibodies with selected binding profiles may be used to assess a level of a protein, a localization pattern of a protein ⁇ e.g., at the tissue, cellular, and/or sub-cellular level) and/or a ratio of a protein with respect to another protein.
  • antibodies with selected binding profiles may be used to assess the level of proMyo statin and the level of latent Myostatin, which may be used to determine a ratio of proMyo statin to latent Myostatin.
  • methods include obtaining one or more antibodies that have a selected binding profile and subjecting a biological sample to an immunoassay using the one or more antibodies with selected binding profiles.
  • 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 or more antibodies having a selected binding profile are used in an assay to assess a biological sample. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 antibodies having a selected binding profile are used in an assay to assess a biological sample.
  • a biological sample is subjected to an immunoassay using two or more antibodies having different selected binding profiles.
  • a biological sample is subjected to an immunoassay using 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 antibodies having different selected binding profiles.
  • antibodies have a selected binding profile to one or more of a human proMyostatin, a human latent Myostatin, a human GDF-8 prodomain, a human GDF-11 ARM8 prodomain, a human proGDFl 1 ARM8, a human mature GDF-8, a human proGDFl 1, and/or a human latent proGDFl 1 ARM8.
  • antibodies have a selected binding profile to one or more of a human proMyostatin, a murine proMyostatin, a human latent Myostatin, a human GDF-8 prodomain, a human GDF-11 ARM8 prodomain, a human proGDFl 1 ARM8, a human mature GDF-8, a human proGDFl 1, a murine latent Myostatin and/or a human latent proGDFl 1 ARM8.
  • the human proMyostatin comprises an amino acid sequence as set forth in SEQ ID NO: 5.
  • the murine proMyostatin comprises an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent Myostatin comprises an amino acid sequence as set forth in SEQ ID NO: 5.
  • the human GDF-8 prodomain comprises an amino acid sequence as set forth in SEQ ID NO: 38.
  • the human GDF- 11 ARM8 prodomain comprises an amino acid sequence as set forth in SEQ ID NO: 181.
  • the human proGDFl 1 ARM8 comprises an amino acid sequence as set forth in SEQ ID NO: 179.
  • the human mature GDF-8 comprises an amino acid sequence as set forth in SEQ ID NO: 42.
  • the human proGDFl 1 comprises an amino acid sequence as set forth in SEQ ID NO: 4.
  • the murine latent Myostatin comprises an amino acid sequence as set forth in SEQ ID NO: 88.
  • the human latent proGDFl 1 ARM8 comprises an amino acid sequence as set forth in SEQ ID NO: 179.
  • a biological sample is subjected to an immunoassay using two or more antibodies having different selected binding profiles.
  • antibodies have selected binding profiles as shown in Table 33.
  • antibodies have selected binding profiles of any one of the antibodies Bla-1, BPlb-1, BP2a- 1, BP3a-l, BP3a-2, BP3a-3, BP4a-l, BP5a-l, BP5b-l, BP5a-2, BP5a-3, BP5a-4, BP5a-5, BP5b-2, BP5b-3, BP5b-4, BP5b-5, BP5b-6, BP5b-7, BP5b-8, BP5b-9, BP6a-l, BP6a-2, BP6a-3, BP6a-4, BP6a-5, BP6a-6, BP6b-l, BP6b-2, BP6a-7, BP6a-8,
  • a biological sample is subjected to one or more
  • immunoassays using a set of two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) antibodies in which the set comprises two antibodies having binding profiles, as outlined in Table 33, selected from the following pairs: ⁇ BPla, BPlb ⁇ , ⁇ BPla, BP2a ⁇ , ⁇ BPla, BP3a ⁇ , ⁇ BPla, BP4a ⁇ , ⁇ BPla, BP5a ⁇ , ⁇ BPla, BP5b ⁇ , ⁇ BPla, BP6a ⁇ , ⁇ BPla, BP6b ⁇ , ⁇ BPla, BP7b ⁇ , ⁇ BPla, BP8b ⁇ , ⁇ BPla, BP9b ⁇ , ⁇ BPla, BPlOb ⁇ , ⁇ BPla, BPl la ⁇ , ⁇ BPla, BP12b ⁇ , ⁇ BPla, BP13a ⁇ , ⁇ BPla, BP14a ⁇ , ⁇ BPla, BP15a ⁇ , ⁇ BPla, BP16b ⁇
  • a biological sample is subjected to an immunoassay using a set of one or more antibodies comprising of any three, four , five, six, seven, eight, nine, ten or more of the antibodies of Table 33.
  • a biological sample is subjected to an immunoassay using three or more antibodies having different selected binding profiles.
  • antibodies have selected binding profiles as shown in Table 33.
  • antibodies have selected binding profiles of any one of the antibodies Bla-1, BPlb-1, BP2a- 1, BP3a-l, BP3a-2, BP3a-3, BP4a-l, BP5a-l, BP5b-l, BP5a-2, BP5a-3, BP5a-4, BP5a-5, BP5b-2, BP5b-3, BP5b-4, BP5b-5, BP5b-6, BP5b-7, BP5b-8, BP5b-9, BP6a-l, BP6a-2, BP6a-3, BP6a-4, BP6a-5, BP6a-6, BP6b-l, BP6b-2, BP6a-7, BP6a-8,
  • methods provided herein relate to methods for detecting one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins in a sample obtained from a subject.
  • a subject refers to an individual organism, for example, an individual mammal.
  • the subject is a human.
  • the subject is a non-human mammal.
  • the subject is a non-human primate.
  • the subject is a rodent (e.g. , a mouse or rat).
  • the subject is a sheep, a goat, a cattle, a cat, or a dog. In some embodiments, the subject is a vertebrate, an amphibian, a reptile, a fish, an insect, a fly, or a nematode. In some
  • the subject is a research animal.
  • the subject is genetically engineered, e.g. , a genetically engineered non-human subject.
  • the subject may be of either sex and at any stage of development.
  • the subject is a patient or a healthy volunteer.
  • a method for detecting one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins in a sample obtained from a subject involves (a) contacting the sample with an antibody having a selected binding profile under conditions suitable for binding of the antibody to the antigen, if the antigen is present in the sample, thereby forming binding complexes; and (b) determining the level of the antibody or antigen binding fragment bound to the antigen (e.g. , determining the level of the binding complexes).
  • a binding complex refers to a biomolecular complex of antibody bound to antigen (e.g. , one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins).
  • Binding complexes may comprise antibodies with a single specificity or two or more antibodies or antigen binding fragments with different specificities.
  • a binding complex comprises two or more antibodies recognizing different antigenic sites on the same antigen.
  • an antibody may be bound to an antigen, having bound to it other biomolecules such as RNA, DNA, polysaccharides or proteins.
  • a binding complex comprises two or more antibodies recognizing different antigens.
  • an antibody in a binding complex e.g., an immobilized antibody bound to antigen
  • an antibody may itself by bound, as an antigen, to an antibody (e.g., a detectably labeled antibody).
  • binding complexes may, in some instances, comprise multiple antigens and multiple antibodies or antigen binding fragments.
  • binding complexes may or may not be in their native in situ conformation.
  • a binding complex is formed between an antibody and a purified protein antigen, or isolated proteins comprising antigen, in which the antigen is not in its native in situ conformation.
  • a binding complex is formed between an antibody and a purified protein antigen, in which the antigen is not in its native in situ conformation and is immobilized on solid support (e.g., a PVDF membrane).
  • a binding complex is formed with an antibody and, for example, a cell surface protein that is present in situ in a native confirmation (e.g., on the surface of a cell).
  • binding complexes may or may not be detectably labeled.
  • binding complexes comprise detectably labeled antibodies and non-labeled antibodies.
  • binding complexes comprise detectably labeled antigen.
  • antibodies, in binding complexes are immobilized to one or more solid supports.
  • antigens, in binding complexes are immobilized to one or more solid supports. Exemplary solid supports are disclosed herein and will be apparent to one of ordinary skill in the art. The foregoing examples of binding complexes are not intended to be limiting. Other examples of binding complexes will be apparent to one or ordinary skill in the art.
  • antibody or antigen may be conjugated to a solid support surface, either directly or indirectly.
  • Methods for conjugation to solid supports are standard and can be accomplished via covalent and non- covalent interactions.
  • Non-limiting examples of conjugation methods include: adsorption, cross-linking, protein A/G - antibody interactions, and streptavidin-biotin interactions. Other methods of conjugation will be readily apparent to one of ordinary skill in the art.
  • detection, diagnosis, and monitoring methods include comparing the level of the antibody (including antigen binding fragments) bound to the antigen (e.g., one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins) to one or more reference standards.
  • the reference standard may be, for example, the level of a corresponding TGFP family member protein in a subject that does or does not have the TGFP family member protein.
  • the reference standard is the level of a TGFP family member protein detected in a sample that does not contain the TGFP family member protein (e.g. , a background level).
  • a background level can be determined from a sample that contains a particular pro/latent-Myostatin, by contacting the sample with non-specific antibodies (e.g. , antibodies obtained from non-immune serum).
  • the reference standard may be the level of a TGFP family member protein detected in a sample that does contain the TGFP family member protein (e.g. , a positive control).
  • the reference standard may be a series of levels associated with varying concentrations of a TGFP family member protein in a sample and useful for quantifying the concentration of the TGFP family member protein in the test sample.
  • the foregoing examples of reference standards are not limiting and other suitable reference standard will be readily apparent to one of ordinary skill in the art.
  • the level of one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins may be measured, as provided herein, from a biological sample using any of the antibodies having a selected binding profile.
  • a biological sample refers to any biological material which may be obtained from a subject or cell.
  • a biological sample may be whole blood, plasma, serum, saliva, cerebrospinal fluid, urine, cells (or cell lysate) or tissue (e.g. , normal tissue or diseased tissue).
  • a biological sample is a fluid sample.
  • a biological sample is a solid tissue sample.
  • a tissue sample may include, without limitation skeletal muscle, cardiac muscle, adipose tissue as well as tissue from other organs.
  • a biological sample is a biopsy sample.
  • a solid tissue sample may be made into a fluid sample using routine methods in the art.
  • a biological sample may also include one or more cells of a cell line.
  • a cell line includes human cells, primate cells (e.g. , vero cells), rat cells (e.g. , GH3 cells, OC23 cells) or mouse cells (e.g. , MC3T3 cells).
  • human cell lines including, without limitation, human embryonic kidney (HEK) cells, HeLa cells, cancer cells from the National Cancer Institute's 60 cancer cell lines (NCI60), DU145 (prostate cancer) cells, Lncap (prostate cancer) cells, MCF-7 (breast cancer) cells, MDA-MB-438 (breast cancer) cells, PC3 (prostate cancer) cells, T47D (breast cancer) cells, THP- 1 (acute myeloid leukemia) cells, U87 (glioblastoma) cells, SHSY5Y human neuroblastoma cells (cloned from a myeloma) and Saos-2 (bone cancer) cells.
  • HEK human embryonic kidney
  • HeLa cells cancer cells from the National Cancer Institute's 60 cancer cell lines (NCI60)
  • DU145 (prostate cancer) cells Lncap (prostate cancer) cells
  • MCF-7 breast cancer
  • MDA-MB-438 breast cancer
  • PC3 prostate cancer
  • a further embodiment relates to a method for monitoring a disease, a condition, or any treatment thereof (e.g., myopathy or myopathy treatment) in a subject having, or at risk of having, the disease or condition comprising: (a) obtaining a biological sample from the subject, (b) determining the level of one or more TGFP family member proteins or forms thereof, one or more portions or domains of TGFP family member proteins and/or one or more chimeras of TGFP family member proteins in the biological sample using an antibody having a selected binding profile and (c) repeating steps (a) and (b) on one or more occasions.
  • a disease, a condition, or any treatment thereof e.g., myopathy or myopathy treatment
  • Biomarkers such as Myostatin may be used as for assessing disease conditions, however, available commercial reagents (e.g., antibodies for ELISAs and Western Blots) are often either not specific for Myostatin, detect only mature Myostatin or do not detect
  • the level of proMyostatin or latent Myostatin may be measured in a subject, or biological sample therefrom, to detect or monitor the progression of a disease or condition.
  • the level of proMyostatin or latent Myostatin may be measured in a subject, or biological sample therefrom, to monitor the response to a treatment for a disease or condition. It should be appreciated that the level of proMyostatin or latent Myostatin may be monitored over any suitable period of time, which may differ depending on the disease or condition, the subject has or any treatment regimen that the subject may be subject to.
  • Another embodiment relates to a diagnostic composition
  • a diagnostic composition comprising any one of the above described antibodies, antigen binding fragments, polynucleotides, vectors or cells and optionally suitable means for detection.
  • the antibodies are, for example, suited for use in immunoassays in which they can be utilized in liquid phase or bound to a solid phase carrier.
  • immunoassays which can utilize the antibody are competitive and noncompetitive immunoassays in either a direct or indirect format. Examples of such
  • immunoassays are the Enzyme Linked Immunoassay (ELISA), radioimmunoassay (RIA), the sandwich (immunometric assay), flow cytometry, the western blot assay,
  • the antigens and antibodies can be bound to many different solid supports (e.g. , carriers, membrane, columns, proteomics array, etc.).
  • solid support materials include glass, polystyrene, polyvinyl chloride, polyvinylidene difluoride, polypropylene, polyethylene, polycarbonate, dextran, nylon, amyloses, natural and modified celluloses, such as nitrocellulose, polyacrylamides, agaroses, and magnetite.
  • the nature of the support can be either fixed or suspended in a solution (e.g. , beads).
  • antibodies may also be used in a method for evaluating expression of one or more TGFP family member proteins or forms thereof in a subject by obtaining a biological sample from the subject which may be a tissue sample, a blood sample or any other appropriate body fluid sample.
  • the procedure may comprise contacting the blood sample (whole blood, serum, plasma), a tissue sample, or protein sample isolated therefrom, with an antibody having a selected binding profile, under conditions enabling the formation of binding complexes between antibody and antigen.
  • the level of such binding complexes may then be determined by any suitable method.
  • the biological sample is contacted with the antibody under conditions suitable for binding of the antibody to one or more TGFP family member proteins or forms thereof, if the antigen is present in the sample, and formation of binding complexes consisting of antibody, bound to the antigen.
  • this contacting step is performed in a reaction chamber, such as a tube, plate well, membrane bath, cell culture dish, microscope slide, and the like.
  • an antibody having a selected binding profile is immobilized on a solid support.
  • the antigen is immobilized on a solid support.
  • the solid support is the surface of a the reaction chamber.
  • the solid support is of a polymeric membrane (e.g. , nitrocellulose strip, Polyvinylidene Difluoride (PVDF) membrane, etc.). Other appropriate solid supports may be used.
  • an antibody having a selected binding profile is
  • immobilized on the solid support prior to contacting with the antigen In other embodiments, immobilization of the antibody is performed after formation of binding complexes. In still other embodiments, antigen is immobilized on a solid support prior to formation of binding complexes.
  • a detection reagent is added to the reaction chamber to detect immobilized binding complexes. In some embodiments, the detection reagent comprises a detectably labeled secondary antibody directed against the antigen. In some embodiments, the primary antibody is itself detectable labeled, and is thereby the detection reagent.
  • detection methods comprise the steps of immobilizing antibodies to a solid support; applying a sample (e.g. , a biological sample or isolated protein sample) to the solid support under conditions that permit binding of antigen to the antibodies, if present in the sample; removing the excess sample from the solid support; applying detectably labeled antibodies under conditions that permit binding of the detectably labeled antibodies to the antigen -bound immobilized antibodies; washing the solid support and assaying for the presence of label on the solid support.
  • a sample e.g. , a biological sample or isolated protein sample
  • the antigen is immobilized on the solid support, such as a PVDF membrane, prior to contacting with the antibody in a reaction chamber (e.g. , a membrane bath).
  • a detection reagent is added to the reaction chamber to detect immobilized binding complexes.
  • the detection reagent comprises a detectably labeled secondary antibody directed against the antigen.
  • the detection reagent comprises a detectably labeled secondary antibody directed against the primary antibody.
  • the detectable label may be, for example, a radioisotope, a fluorophore, a luminescent molecule, an enzyme, a biotin-moiety, an epitope tag, or a dye molecule.
  • the primary antibody is itself detectable labeled, and is thereby the detection reagent. Suitable detectable labels are described herein, and will be readily apparent to one of ordinary skill in the art.
  • diagnostic kits suitable for home or clinical use (point of care service), comprise (a) detectably labeled and/or non-labeled antibodies having a selected binding profile, as antigen binding reagents (e.g. , reagents that bind one or more TGFP family member proteins or forms thereof); (b) a detection reagent; and, optionally, (c) complete instructions for using the reagents to detect antigens in a sample.
  • the diagnostic kit includes the antibody having a selected binding profile, and/or one or more TGFP family member proteins or forms thereof immobilized on a solid support. Any of the solid supports described herein are suitable for incorporation in the diagnostic kits.
  • the solid support is the surface of a reaction chamber of a plate well.
  • the plate well is in a multi-well plate having a number of wells selected from: 6, 12, 24, 96, 384, and 1536, but it is not so limited. In other words,
  • the diagnostic kits provide a detectably labeled antibody. Diagnostic kits are not limited to these embodiments and other variations in kit composition will be readily apparent to one of ordinary skill in the art.
  • Myostatin protein is present in muscle tissue. In some embodiments, Myostatin protein is present in circulation. In some embodiments, Myostatin protein is present in muscle tissue and in circulation. In some embodiments, Myostatin expression is altered in diseases or conditions involving muscle atrophy. Exemplary diseases or conditions involving muscle atrophy include, without limitation, muscular dystrophy, cachexia, sarcopenia, spinal cord injury and atrophy associated with muscle disuse. Thus, Myostatin in tissues or circulating Myostatin, can be a biomarker for muscular atrophic disease.
  • Myostatin as biomarker has been hampered by the paucity of available antibodies to Myostatin, which, because of the high homology between mature Myostatin growth factor and other members of the TGFP family, are not highly selective for Myostatin. Furthermore, many Myostatin antibodies cannot distinguish between the pro-, latent, and mature forms of Myostatin. Thus, detecting the location and expression levels of various Myostatin forms (e.g., proMyostatin, latent Myostatin, primed Myostatin, and mature Myostatin) may provide significant insight into Myostatin biology in normal and disease states.
  • various Myostatin forms e.g., proMyostatin, latent Myostatin, primed Myostatin, and mature Myostatin
  • antibodies to pro-, latent, and mature Myostatin may be useful in biomarker assays for muscle atrophy.
  • antibodies provided herein can be incorporated into assays for MSD, ELISA, RIA, quantitative western blotting (capillary western, Odyssey or similar technology), immunohistochemistry, and any other technology requiring specific binding of antibodies.
  • assays are be applied to analysis of Myostatin levels and isoform distribution in muscle or other tissue, blood and blood derivatives (e.g., serum, plasma, exosome-like vesicles (ELVs) from blood), and urine and derivatives (e.g., urinary ELVs).
  • blood and blood derivatives e.g., serum, plasma, exosome-like vesicles (ELVs) from blood
  • urine and derivatives e.g., urinary ELVs.
  • any of the antibodies provided herein can be used in an immunohistochemistry (IHC) assay.
  • an immunohistochemistry assay is performed on a tissue sample (e.g., a muscle biopsy) to identify a level or localization of one or more TGFP family member proteins or forms thereof (e.g., proMyostatin, latent Myostatin , primed Myostatin, and/or mature Myostatin).
  • a tissue sample e.g., a muscle biopsy
  • TGFP family member proteins or forms thereof e.g., proMyostatin, latent Myostatin , primed Myostatin, and/or mature Myostatin.
  • an IHC assay begins with antigen retrieval, which may vary in terms of reagents and methods.
  • the antigen retrieval process may involve pressure cooking, protease treatment, microwaving, or heating histologic sections in baths of appropriate buffers, with the standard goal of unmasking antigens hidden by formalin crosslinks or other fixation. See, e.g., Leong et al. Appl. Immnunohistochem. 4(3):201 (1996).
  • an IHC assay is a direct assay.
  • an IHC assay is an indirect assay.
  • binding of antibody to the target antigen can be determined directly.
  • a direct assay may use a labeled reagent, such as a fluorescent tag or an enzyme-labeled primary antibody, which can be visualized without further antibody interaction.
  • unconjugated primary antibody binds to the antigen and then a labeled secondary antibody binds to the primary antibody.
  • a chromagenic or fluorogenic substrate is added to provide visualization of the antigen. Signal amplification may occur because several secondary antibodies may react with different epitopes on the primary antibody.
  • a primary and/or secondary antibody used for IHC is labeled with a detectable moiety.
  • labels include, without limitation, radioisotopes, colloidal gold particles, fluorescent or chemiluminescent labels.
  • a primary and/or secondary antibody used for IHC is labeled with a, radioisotope (e.g. , S, C, I or H) 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.
  • radionuclides include, without limitation, 99Tc,90Y, mln, 32P, UC, 150, 13N, 18F, 51Cr, 57To, 225Ra, 60Co, 59Fe, 57Se, 152Eu, 67CU, 217Ci, 211 At, 212Pb, 47Sc, 109Pd, 234Th, and 40K, 157Gd, 55Mn, 52Tr, and 56Fe.
  • a primary and/or secondary antibody used for IHC is labeled with Colloidal gold particles.
  • a primary and/or secondary antibody used for IHC is labeled with fluorescent or chemiluminescent labels including, but not limited to, rare earth chelates (europium chelates), fluorescein and its derivatives, rhodamine and its derivatives, isothiocyanate, phycoerythrin, phycocyanin, allophycocyanin, o-phthaladehyde, fluorescamine, dansyl, umbelliferone, luciferin, luminal label, isoluminal label, an aromatic acridinium ester label, an imidazole label, an acridimium salt label, an oxalate ester label, an aequorin label, 2,3- dihydrophthalazinediones, Texas Red, dansyl, Lissamine, umbelliferone, phycocrytherin, phycocyanin, or commercially available fluorophores such SPECTRUM ORANGE® and SPECTRUM
  • a primary and/or secondary antibody used for IHC is labeled with an enzyme.
  • an enzyme catalyzes a chemical alteration of a chromogenic substrate that can be measured using various techniques. For example, an enzyme may catalyze a color change in a substrate, which can be measured
  • an enzyme may alter the fluorescence or
  • chemiluminescence of the substrate involves techniques for quantifying a change in fluorescence would be apparent to the skilled artisan.
  • a chemiluminescent substrate becomes electronically excited by a chemical reaction and may then emit light which can be measured (e.g., using a chemiluminometer) or donates energy to a fluorescent acceptor.
  • enzymatic labels include luciferases (e.g. firefly luciferase and bacterial luciferase; U.S. Pat. No. 4,737,456), luciferin, 2,3-dihydrophthalazinediones, malate dehydrogenase, urease, peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase, ⁇ -galactosidase, glucoamylase, lysozyme, saccharide oxidases (e.g., glucose oxidase, galactose oxidase, and glucose-6-phosphate dehydrogenase), heterocyclic oxidases (such as uricase and xanthine oxidase), lactoperoxidase, microperoxidase, and the like.
  • luciferases e.g. firefly luciferase and bacterial luciferase
  • a label is indirectly conjugated to any of the antibodies with a selected binding profile provided herein.
  • an antibody can be conjugated with biotin and any suitable label 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.
  • a tissue section e.g., an FFPE section or a frozen section
  • a tissue section is exposed to one or more of the antibodies provided herein as primary antibodies for a sufficient period of time and under suitable conditions to allow one or more primary antibody to bind to one or more TGFP family member proteins or forms thereof of the tissue section.
  • suitable conditions for achieving suitable binding can be determined by routine experimentation,
  • a slide containing a biological sample is subjected to a wash step to remove unbound and excess amounts of primary antibody.
  • primary antibody is linked to a detectable label, such as paramagnetic ions, radioactive isotopes, fluorochromes, and NM -detectable substances, and the slide is evaluated for staining of one or more TGFP family member proteins or forms thereof using an appropriate imaging apparatus.
  • a detectable label such as paramagnetic ions, radioactive isotopes, fluorochromes, and NM -detectable substances
  • tissue sample used to prepare stained tissue sections for evaluating expression of one or more TGFP family member proteins or forms thereof can be collected from a subject before and/or after exposure of the subject to one or more therapeutic agents, e.g., an anti-latent Myostatin antibody. Accordingly, tissue samples may be collected from a subject over a period of time.
  • a tissues sample e.g., a muscle tissue sample
  • a tissue sample is obtained by any suitable procedure, which includes, but not limited to, surgical excision, aspiration or biopsy.
  • a tissue sample is sectioned and examined for one or more TGFP family member proteins or forms thereof as a fresh specimen.
  • a tissue sample is frozen for further sectioning.
  • a tissue sample is preserved by fixing and embedding in paraffin (FFPE) or the like.
  • a tissue sample may be fixed by conventional methodology, with the length of fixation depending on the size of the tissue sample and the fixative used.
  • fixatives include, but are not limited to neutral buffered formalin, glutaraldehyde, Bouin's or paraformaldehyde.
  • a tissue sample is fixed with formalin.
  • a fixed tissue sample is also embedded in paraffin to prepare a formalin-fixed and paraffin- embedded (FFPE) tissue sample.
  • FFPE formalin-fixed and paraffin- embedded
  • a tissue sample is fixed and dehydrated through an ascending series of alcohols, infiltrated and embedded with paraffin or other sectioning media so that the tissue sample may be sectioned.
  • a tissue sample is first sectioned and then the individual sections are fixed.
  • Growth factors are cell signaling molecules that stimulate a variety of cellular activities. Due to their broad-reaching influence within biological systems, growth factor signaling is tightly regulated, often through interactions with other biomolecules, the extracellular and/or cellular matrix or within a particular cell environment or niche. These interactions may be direct or indirect.
  • TGF- ⁇ transforming growth factor beta
  • Activated type I receptors may in turn phosphorylate receptor-associated SMADs (R-SMADs) promoting co-SMAD (e.g. SMAD4) dimer/trimer formation and nuclear translocation.
  • R-SMADs receptor-associated SMADs
  • co-SMAD e.g. SMAD4 dimer/trimer formation and nuclear translocation.
  • SMAD complexes collaborate with cofactors to modulate expression of TGF- ⁇ family member target genes.
  • TGF- ⁇ family member signaling cascades are involved in a number of diverse biological pathways including, but not limited to inhibition of cell growth, tissue
  • TGF- ⁇ signaling related to growth inhibition and tissue homeostasis may affect epithelial, endothelial, hematopoietic and immune cells through the activation of p21 and p 15 to mediate cell cycle arrest and repress myc.
  • ECM remodeling TGF- ⁇ signaling may increase fibroblast populations and ECM deposition (e.g. collagen).
  • TGF- ⁇ signaling related to cell migration and invasion may affect epithelial and/or endothelial cells, inducing stem cell-like phenotypes. This aspect of signaling may play a role in smooth muscle cell proliferation following vascular surgery and/or stenting.
  • TGF- ⁇ ligand is necessary for T regulatory cell function and maintenance of immune precursor cell growth and homeostasis.
  • Nearly all immune cells comprise receptors for TGF- ⁇ and TGF- ⁇ knockout mice die postnataly due in part to inflammatory pathologies.
  • TGF- ⁇ suppresses interferon gamma-induced activation of natural killer cells (Wi, J. et al., 2011. Hepatology. 53(4): 1342-51, the contents of which are herein incorporated by reference in their entirety).
  • TGF-beta The solution of the crystal structure of the latent form of TGF-beta is a first for the entire TGF-beta family and offers deep insights into these complexes (Shi, M. et al., Latent TGF- ⁇ structure and activation. Nature. 2011 Jun 15; 474(7351):343-9). Almost all signaling in the TGF-beta family goes through a common pathway whereby a dimeric ligand is recognized by a heterotetrameric receptor complex containing two type I and two type II receptors. Each receptor has a serine-threonine kinase domain. Type II receptors
  • phosphorylate type I receptors which in turn phosphorylate receptor-regulated Smads that translocate to and accumulate in the nucleus and regulate transcription.
  • BMP bone morphogenetic proteins
  • GDF growth and differentiation factor
  • myostatin nodal, anti-Mullerian hormone, and lefty proteins.
  • mature growth factors are synthesized along with their prodomains as single polypeptide chains (see FIG. 2).
  • polypeptide chains may comprise cleavage sites for separation of prodomains from mature growth factors.
  • cleavage sites are furin cleavage sites recognized and cleaved by proprotein convertases.
  • prodomain homology In general, homology among TGF- ⁇ family member growth factor domains is relatively high. Interestingly, prodomain homology is much lower. This lack of homology may be an important factor in altered growth factor regulation among family members. In some cases, prodomains may guide proper folding and/or dimerization of growth factor domains. Prodomains have very recently been recognized, in some cases, to have important functions in directing growth factors (after secretion) to specific locations in the extracellular matrix (ECM) and/or cellular matrix, until other signals are received that cause growth factor release from latency. Release from latency may occur in highly localized environments whereby growth factors may act over short distances (e.g.
  • growth factor-prodomain complexes are secreted as homodimers.
  • prodomain-growth factor complexes may be secreted as heterodimers.
  • TGF-P-related protein refers to a TGF- ⁇ isoform, a TGF- ⁇ family member or a TGF- ⁇ family member-related protein.
  • TGF- ⁇ family members may include, but are not limited to any of those shown in in FIG. 1 and/or listed in Table 1. These include, but are not limited to TGF- ⁇ proteins, BMPs, myostatin, GDFs and inhibins.
  • Aspects of the present disclosure provide tools and/or methods for characterizing and/or modulating cellular activities related to growth factor signaling.
  • tools of the present disclosure may comprise antigens comprising one or more components of one or more TGF ⁇ -related proteins.
  • tools of the present disclosure may comprise antibodies directed toward antigens of the present disclosure.
  • tools of the present disclosure may comprise assays for the detection and/or characterization of TGF ⁇ -related proteins, the detection and/or characterization of antibodies directed toward TGF ⁇ -related proteins and/or the detection and/or characterization of cellular activities and/or their cellular signaling related to TGF ⁇ -related proteins.
  • GDF growth differentiation factor
  • TGF- ⁇ family member proteins involved in a number of cellular and developmental activities.
  • GDF-modulatory antibodies are capable of distinguishing between various growth factor complexes allowing for growth factor activity modulation that occurs only at sites of specific complex formations.
  • TGF ⁇ -related proteins are involved in a number of cellular processes. In embryogenesis, the 33 members of the TGF- ⁇ family of proteins are involved in regulating major developmental processes and the details of the formation of many organs. Much of this regulation occurs before birth; however, the family continues to regulate many processes after birth, including, but not limited to immune responses, wound healing, bone growth, endocrine functions and muscle mass. TGF-P-related proteins are listed and described in International Patent Application No. WO2014074532, the contents of which are herein incorporated by reference in their entirety.
  • TGF- ⁇ family pro-proteins i.e. the protein after removal of the secretion signal sequence
  • the pro-protein contains, and is the precursor of, the prodomain and the growth factor. Shown in the Table are the names of the originating TGF- ⁇ family member and the pro-protein sequence. Also identified in "bold" and
  • proprotein convertase cleavage sites underlined are proprotein convertase cleavage sites. Upon cleavage, the resulting prodomain retains this site, whereas the mature growth factor begins following the cleavage site. It is noted that Leftyl and Lefty2 are not cleaved by proprotein convertases just prior to the start of the mature growth factor.
  • TGF- 2 SLSTCSTLDMDQFMRKRIEAIRGQILSKLKLTSPPEDYPEPEEV 2
  • GDF-8 NENSEQKENVEKEGLCNACTWRQNTKSSRIEAIKIQILSKLRL 5 (myostatin) ETAPNISKDVIRQLLPKAPPLRELIDQYDVQRDDSSDGSLEDD
  • GDF-7 RDGLEAAAVLRAAGAGPVRSPGGGGGGGGRTLAQAAGA 19
  • proprotein convertase refers to an enzyme that cleaves a prodomain from a translated protein to facilitate protein maturation.
  • Some proprotein convertases of the present disclosure include the subtilisin-like proprotein convertase (SPC) family member enzymes.
  • SPC family comprises calcium-dependent serine endopro teases that include, but are not limited to furin/PACE, PC 1/3, PC2, PC4, PC5/6, PACE4 and PC7 (Fuller et al., 2009. Invest Ophthalmol Vis Sci. 50(12):5759-68, the contents of which are herein incorporated by reference in their entirety).
  • GDF-11 may in some cases, be cleaved by PC5/6.
  • proprotein convertases may cleave proproteins at additional sites, other than those indicated in Table 1.
  • pro-proteins may be cleaved at a first cleavage site (the first site being the site closest to the N-terminus).
  • pro-proteins may be cleaved at a cleavage site other than a first cleavage site.
  • proprotein convertase cleavage may occur intracellularly.
  • proprotein convertase cleavage may occur extracellularly.
  • TGF- ⁇ family member proteins are synthesized in conjunction with prodomains. Some prodomains may remain associated with growth factors after cleavage. Such associations may form latent growth factor-prodomain complexes (GPCs) that modulate the availability of growth factors for cell signaling. Growth factors may be released from latency in GPCs through associations with one or more extracellular proteins. In some cases, growth factor release may rely on force applied to GPCs through extracellular protein interactions. Such forces may pull from C-terminal and/or N-terminal regions of GPCs resulting in the release of associated growth factors.
  • GPCs latent growth factor-prodomain complexes
  • the prodomain portion of the GPC is responsible for growth factor retention and blocking the interaction of retained growth factors with their receptors.
  • Such GPCs where the bound growth factor is unable to promote signaling activity, are also referred to herein as "latent complexes.”
  • Prodomain portions of GPCs that function to block growth factor signaling activity are referred to as latency associated peptides (LAPs).
  • LAPs latency associated peptides
  • TGF- ⁇ , 2 and 3 are know to comprise LAPs.
  • GDF prodomains also function to block growth factor activity.
  • Some prodomains may comprise LAP-like domains.
  • LAP-like domain refers to prodomain portions of GPCs and/or free prodomains that may be structurally similar or synthesized in a similar manner to LAPs, but that may not function to prevent growth factor/receptor interactions.
  • FIG. 3 is a schematic depicting an embodiment of a GPC before and after growth factor release.
  • growth factor dimers may associate with prodomain modules to form a GPC.
  • GPCs comprise protein modules necessary for different aspects of growth factor signaling, secretion, latency and/or release from latent GPCs.
  • protein module refers to any component, region and/or feature of a protein. Protein modules may vary in length, comprising one or more amino acids.
  • Protein modules may be from about 2 amino acid residues in length to about 50 amino acid residues in length, from about 5 amino acid residues in length to about 75 amino acid residues in length, from about 10 amino acid residues in length to about 100 amino acid residues in length, from about 25 amino acid residues in length to about 150 amino acid residues in length, from about 125 amino acid residues in length to about 250 amino acid residues in length, from about 175 amino acid residues in length to about 400 amino acid residues in length, from about 200 amino acid residues in length to about 500 amino acid residues in length and/or at least 500 amino acid residues in length.
  • protein modules comprise one or more regions with known functional features (e.g. protein binding domain, nucleic acid binding domain, hydrophobic pocket, etc). Protein modules may comprise functional protein domains necessary for different aspects of growth factor signaling, secretion, latency and/or release from latent conformations.
  • protein modules may be derived from TGF-P-related proteins.
  • Such protein modules may include, but are not limited to prodomains (e.g. latency- associated peptides (LAPs) or LAP-like domains), growth factor domains, fastener regions, proprotein convertase cleavage sites (e.g. furin cleavage sites), B/TP cleavage sites, arm regions, finger regions, residues (such as cysteine residues for example) for extracellular protein associations, latency loops (also referred to herein as latency lassos), alpha 1 helical regions, alpha 2 helical regions, RGD sequences and bowtie regions.
  • FIG. 4 is a schematic diagram showing some of the protein modules present in a GPC as well as in the prodomains and growth factor dimer after activating cleavage of the GPC.
  • prodomains may associate with growth factors in GPCs. Some prodomains may sterically prevent growth factor association with one or more cellular receptors. Prodomains may comprise arm regions and/or straight jacket regions. Some prodomains may comprise C-terminal regions referred to herein as "bowtie regions.” In some prodomain dimers, bowtie regions of each monomer may associate and/or interact. Such associations may comprise disulfide bond formation, as is found between monomers of TGF- ⁇ isoform LAPs.
  • arm regions may comprise trigger loop regions.
  • Trigger loops may comprise regions that associate with integrins. Such regions may comprise amino acid sequences comprising RGD (Arg-Gly-Asp). Regions comprising RGD sequences are referred to herein as RGD sequence regions.
  • prodomains comprise latency loops (also referred to herein as latency lassos). Some latency loops may maintain associations between prodomains and growth factors present within GPCs.
  • Prodomains may also comprise fastener regions. Such fastener regions may promote associations between prodomains and growth factors present within GPCs by maintaining prodomain
  • GPCs may require enzymatic cleavage to promote dissociation of bound growth factors and growth factor activity. Such enzymatic cleavage events are referred to herein as "activating cleavage" events. Activating cleavage of GPCs may be carried out in some instances by members of the BMP- 1/Tolloid- like proteinase (B/TP) family (Muir et al., 2011. J Biol Chem. 286(49):41905-11, the contents of which are herein incorporated by reference in their entirety).
  • B/TP BMP- 1/Tolloid- like proteinase
  • metaloproteinases may include, but are not limited to BMP- 1, mammalian tolloid protein (mTLD), mammalian tolloid-like 1 (mTLLl) and mammalian tolloid-like 2 (mTLL2).
  • Exemplary GPCs that may be subjected to activating cleavage by such metalloproteinases may include, but are not limited to GDF-8 and GDF-11.
  • GDF-8 may be cleaved by mTLL2.
  • activating cleavages may occur intracellularly. In some cases, activating cleavages may occur extracellularly.
  • GDF-8 and GDF-11 GPCs may be transformed by furin cleavage into a latent complex that further requires cleavage by
  • the present disclosure provides polypeptide inhibitors (e.g., antibodies) that inhibit one or more members of the B/TP family. Such inhibitors may block cleavage of BMP-l/Tolloid cleavage sites, including, but not limited to BMP/Tolloid cleavage sites on one or more latent complexes (e.g., GDF-8 latent complexes and/or GDF-11 latent complexes).
  • polypeptide inhibitors e.g., antibodies
  • Such inhibitors may block cleavage of BMP-l/Tolloid cleavage sites, including, but not limited to BMP/Tolloid cleavage sites on one or more latent complexes (e.g., GDF-8 latent complexes and/or GDF-11 latent complexes).
  • activating cleavage may not lead to dissociation of bound growth factor, but instead may promote an active conformation of the GPC.
  • active conformation refers to a GPC protein confirmation that allows the growth factor to engage in receptor interaction.
  • proBMP-7 and proBMP-9 See FIG. 1 .
  • Active conformations of GDF-8 GPCs or GDF-11 GPCs are referred to herein as "primed" complexes, and can be produced by the sequential cleavage of GPCs at the furin cleavage site and the BMP/Tolloid cleavage site.
  • Primed complexes (either the entire complex or portions of the complex) may bind receptors resulting in receptor signaling.
  • prodomains may be dissociated from growth factors upon receptor binding and/or signaling activity.
  • prodomains may remain associated with growth factors upon receptor binding and/or signaling activity.
  • prodomains may become partially dissociated from growth factors during receptor binding and/or signaling activity.
  • primed complexes may bind preferentially to one or more receptors over one or more other receptors.
  • receptor activity resulting from primed complex interactions may be quenched or competed for by excess prodomain or fragments thereof.
  • the present disclosure provides polypeptide inhibitors (e.g., inhibiting antibodies) that block the formation of primed complexes from latent complexes.
  • such inhibitors bind BMP/Tolloid cleavage sites on latent GPCs (e.g., latent GDF-8 or latent GDF-11).
  • such inhibitors prevent cleavage of the BMP/Tolloid cleavage site.
  • Straight jacket regions may comprise alpha 1 helical regions.
  • alpha 1 helical regions may be positioned between growth factor monomers. Some alpha 1 helical regions comprise N-terminal regions of prodomains. Alpha 1 helical regions may also comprise N-terminal regions for extracellular associations. Such extracellular associations may comprise extracellular matrix proteins and/or proteins associated with the extracellular matrix. Some extracellular associations may comprise associations with proteins that may include, but are not limited to LTBPs (e.g. LTBP1, LTBP2, LTBP3 and/or LTBP4), fibrillins (e.g. fibrillin- 1, fibrillin-2, fibrillin-3 and/or fibrillin-4), perlecan, decorin and/or GASPs. N-terminal extracellular associations may comprise disulfide bonds between cysteine residues. In some cases, extracellular matrix proteins and/or proteins associated with the extraceullar matrix may comprise bonds or interactions with one or more regions of prodomains other than N-terminal regions.
  • LTBPs e.g. LTBP1, LTBP2, LTBP3
  • growth factor domains comprise one or more growth factor monomers. Some growth factor domains comprise growth factor dimers. Such growth factor domains may comprise growth factor homodimers or heterodimers (comprising growth factor monomers from different TGF-P-related proteins). Some growth factor domains may comprise fingers regions. Such fingers regions may comprise ⁇ -pleated sheets. Fingers regions may associate with prodomains. Some fingers regions may maintain association between growth factor domains and prodomains.
  • recombinant proteins of the present disclosure may comprise protein modules from growth differentiation factor (GDF) proteins.
  • GDF protein modules may comprise the protein modules and/or amino acid sequences listed in Table 2.
  • protein modules of the present disclosure may comprise amino acid sequences similar to those in Table 2, but comprise additional or fewer amino acids than those listed.
  • Some such amino acid sequences may comprise about 1 more or fewer amino acids, about 2 more or fewer amino acids, about 3 more or fewer amino acids, about 4 more or fewer amino acids, about 5 more or fewer amino acids, about 6 more or fewer amino acids, about 7 more or fewer amino acids, about 8 more or fewer amino acids, about 9 more or fewer amino acids, about 10 more or fewer amino acids or greater than 10 more or fewer amino acids on N-terminal and/or C-terminal ends.
  • GDF-8 fingers region 1 CRYPLTVDFEAFGWDWIIAPKRYKANYCS 47 and GDF- 11
  • Some recombinant proteins of the present disclosure may comprise GDF-15, GDF- 15 signaling pathway-related proteins and/or modules and/or portions thereof.
  • GDF-15 is a TGF- ⁇ family protein that is highly expressed in liver. Expression of GDF-15 is dramatically upregulated following liver injury (Hsiao et al. 2000. Mol Cell Biol. 20(10):3742-51).
  • GDF-15 comprises less than 30% homology with other members, making it the most divergent member of the family (Tanno et al., 2010. Curr Opin Hematol. 17(3): 184-90, the contents of which are incorporated herein by reference in their entirety).
  • the mature form is soluble and can be found in the blood stream.
  • GDF-15 levels in circulation have been found to negatively correlate with hepcidin levels, suggesting a role for GDF-15 in iron load and/or metabolism (Finkenstedt et al., 2008. British Journal of Haematology. 144:789-93). Elevated GDF-15 in the blood is also associated with ineffective and/or apoptotic erythropoiesis, such as in subjects suffering from beta-thalassemia or dyserythropoietic anemias.
  • recombinant proteins of the present disclosure may comprise protein modules from activin subunits.
  • Such protein modules may comprise the protein modules and/or amino acid sequences of the activin subunit inhibin beta A, listed in Table 3.
  • protein modules of the present disclosure may comprise amino acid sequences similar to those in Table 3, but comprise additional or fewer amino acids than those listed.
  • Some such amino acid sequences may comprise about 1 more or fewer amino acids, about 2 more or fewer amino acids, about 3 more or fewer amino acids, about 4 more or fewer amino acids, about 5 more or fewer amino acids, about 6 more or fewer amino acids, about 7 more or fewer amino acids, about 8 more or fewer amino acids, about 9 more or fewer amino acids, about 10 more or fewer amino acids or greater than 10 more or fewer amino acids on N-terminal and/or C-terminal ends.
  • Prodomains may vary in length from about 50 to about 200, from about 100 to about 400 or from about 300 to about 500 amino acids residues. In some embodiments, prodomains range from about 169 to about 433 residues. Prodomains may be unrelated in sequence and/or low in homology. Some prodomains may have similar folds and/or three dimensional structures. Prodomains of TGF- ⁇ family members may comprise latency loops. Such loops may be proline-rich. Latency loop length may determine the ability of such loops to encircle growth factor finger regions.
  • protein modules from some TGF- ⁇ family members comprise low sequence identity with protein modules from other TGF- ⁇ family members. Such low sequence identity may indicate specialized roles for such family members with distinct protein modules.
  • GPCs Association of GPCs with extracellular proteins may strengthen prodomain-growth factor interactions.
  • extracellular proteins may include, but are not limited to LTBPs, fibrillins, GASPs, perlecan and/or decorin.
  • extracellular protein associations are required to keep growth factors latent in GPCs.
  • GARP expression has been shown to be required for surface expression of GPCs on the surface of cells of hematopoietic origin (Tran, D.Q. et al., GARP (LRRC32) is essential for the surface expression of latent TGF- ⁇ on platelets and activated FOXP3+ regulatory T cells. PNAS. 2009, Jun 2. 106(32): 13445-50). GARP may act as a tether to hold GPCs in place on the surface of these cells, including, but not limited to regulatory T-cells and/or platelets.
  • recombinant proteins of the present disclosure may comprise bone morphogenetic proteins (BMPs), a family of TGF-P-related proteins.
  • BMPs bone morphogenetic proteins
  • Protein modules comprising sequences from BMPs may comprise sequences from any of those BMP modules disclosed in FIG. 6. While related to other TGF- ⁇ family member proteins, BMPs generally signal through SMAD1, 5 and 8 proteins while TGF- ⁇ isoforms (e.g. TGF- ⁇ , TGF-P2 and TGF-P3) signal through SMAD2 and SMAD3.
  • Some BMP receptors and/or co-receptors are also distinct from other TGF- ⁇ family member proteins.
  • RGM repulsive guidance molecule
  • RGM proteins act as co-receptors for BMP signaling.
  • RGMA, RGMB and RGMC also known as hemojuvelin (Hjv)].
  • Recombinant proteins of the present disclosure comprising one or more BMP protein module may be useful for the development of antibodies and/or assays to study, enhance and/or perturb BMP interactions with RGM proteins.
  • CTCK domain-containing proteins Another family of GDF/BMP interacting proteins is C-terminal cysteine knot-like (CTCK) domain-containing proteins.
  • CTCK domain-containing proteins may act antagonistically with regard to GDF/BMP signal transduction.
  • CTCK domain-containing proteins include, but are not limited to Cerberus, Connective tissue growth factor (CTGF), DAN domain family member 5 (DAND5), Gremlin- 1 (GREM1), Gremlin-2 (GREM2), Mucin-19 (MUC19), Mucin-2 (MUC2), Mucin-5AC (MUC5AC), Mucin-5B (MUC5B), Mucin-6 (MUC6), Neuroblastoma suppressor of tumorigenicity 1 (NBL1), Norrin (NDP), Otogelin (OTOG), Otogelin-like protein (OTOGL), Protein CYR61 (CYR61), Protein NOV homolog (NOV), Sclerostin (SOST), Sclerostin domain-containing protein 1 (SOSTDC
  • the present disclosure provides recombinant proteins.
  • the term "recombinant protein” refers to a protein produced by an artificial gene and/or process (e.g. genetic engineering). Such recombinant proteins may comprise one or more protein modules from one or more TGF-P-related proteins. Some recombinant proteins disclosed herein may be useful as recombinant antigens.
  • the term “recombinant protein” refers to a protein produced by an artificial gene and/or process (e.g. genetic engineering). Such recombinant proteins may comprise one or more protein modules from one or more TGF-P-related proteins. Some recombinant proteins disclosed herein may be useful as recombinant antigens.
  • the term “recombinant protein” refers to a protein produced by an artificial gene and/or process (e.g. genetic engineering). Such recombinant proteins may comprise one or more protein modules from one or more TGF-P-related proteins. Some recombinant proteins disclosed herein may be useful as recomb
  • recombinant antigen refers to a recombinant protein that may be used to immunize one or more hosts for the production of antibodies directed toward one or more epitopes present on such recombinant antigens. Some recombinant antigens may be cell-based antigens. As used herein, the term "cell-based antigen” refers to recombinant antigens that are expressed in cells for presentation of such antigens on the cell surface. Such cells may be used to immunize hosts for the production of antibodies directed to such cell-based antigens.
  • recombinant proteins disclosed herein may be used as therapeutics.
  • Recombinant proteins disclosed herein may modulate growth factor (e.g. growth factors comprising TGF-P-related proteins) levels and/or activity (e.g. signaling) upon administration and/or introduction to one or more subjects and/or niches.
  • growth factor e.g. growth factors comprising TGF-P-related proteins
  • activity e.g. signaling
  • recombinant proteins disclosed herein may be used to assay growth factor (e.g. growth factors comprising TGF-P-related proteins) levels and/or activity (e.g. signaling). Some recombinant proteins disclosed herein may be used in the isolation of antibodies directed to TGF-P-related proteins. Recombinant proteins of the present disclosure may also be used as recombinant antigens in the development of stabilizing [reducing or preventing dissociation between two agents, (e.g. growth-factor release from GPCs, GPC release from one or more protein interactions)] and/or releasing [enhancing the dissociation between two agents (e.g. growth-factor release from GPCs, GPC release from one or more protein interactions)] antibodies.
  • stabilizing reducing or preventing dissociation between two agents, (e.g. growth-factor release from GPCs, GPC release from one or more protein interactions)] and/or releasing [enhancing the dissociation between two agents (e.g. growth-factor release from GPCs, GPC release from
  • Recombinant proteins of the present disclosure may include TGF- ⁇ family member proteins as well as components and/or protein modules thereof. Some recombinant proteins of the present disclosure may comprise prodomains without associated growth factors, furin cleavage-deficient mutants, mutants deficient in extracellular protein associations and/or combinations thereof.
  • recombinant proteins may comprise detectable labels.
  • Detectable labels may be used to allow for detection and/or isolation of recombinant proteins.
  • Some detectable labels may comprise biotin labels, polyhistidine tags and/or flag tags. Such tags may be used to isolate tagged proteins.
  • Proteins produced may comprise additional amino acids encoding one or more 3C protease cleavage site. Such sites allow for cleavage at the 3C protease cleavage site upon treatment with 3C protease, including, but not limited to rhinovirus 3C protease. Such cleavage sites are introduced to allow for removal of detectable labels from recombinant proteins.
  • FIG. 7 is a schematic depicting an embodiment of a recombinant GPC.
  • Recombinant proteins according to FIG. 7 comprising TGF-P-family member proteins may comprise features including, but not limited to C-terminal regions of the mature growth factor, N-terminal regions of the prodomain and/or proprotein cleavage sites.
  • the proprotein cleavage site of recombinant GPCs may, for example, comprise the furin consensus sequences RXXR, or RXR wherein R is arginine and X indicates amino acid residues that may vary among TGF- ⁇ family members.
  • Furin cleavage site sequences (although not limited to cleavage by furin alone and may include cleavage by other proprotein convertase enzymes) for each TGF- ⁇ family member are indicated in Table 1.
  • prodomains of GPCs may comprise one or more cysteine residues within and/or near the N-terminal region of the prodomain.
  • cysteine residues may be from about 1 to about 10 amino acids, from about 4 to about 15 amino acids, from about 5 to about 20 amino acids and/or from about 7 to about 50 amino acids from the N-terminus of the prodomain.
  • recombinant GPCs may also be expressed with detectable labels. Such detectable labels may be useful for detection and/or isolation of recombinant GPCs. Detectable labels may comprise 2 or more histidine (His) residues. Such detectable labels may also be referred to herein as polyhistidine tags.
  • Polyhistidine tags may include hexa histidine tags or HIS-TAGTM (EMD Biosciences, Darmstadt, Germany) comprising a chain of six histidine residues. Some polyhistidine tags may be present at the N-terminus of recombinant proteins disclosed herein.
  • polyhistidine tags may be present at the C-terminus of recombinant proteins disclosed herein. Proteins produced may comprise additional amino acids encoding one or more 3C protease cleavage site. Such sites allow for cleavage at the 3C protease cleavage site upon treatment with 3C protease, including, but not limited to rhinovirus 3C protease. Some cleavage sites may be introduced to allow for removal of detectable labels from recombinant proteins.
  • recombinant GPCs may comprise mutations in one or more amino acids as compared to wild type sequences.
  • one or more regions of proteolytic processing may be mutated (see FIG. 8).
  • Such regions may comprise proprotein convertase cleavage sites.
  • Proprotein convertase e.g. furin
  • cleavage site mutations prevent enzymatic cleavage at that site and/or prevent enzymatic cleavage of growth factors from their prodomains.
  • Some proprotein convertase cleavage sites comprising RXXR sequences may be mutated to RXG (wherein X indicates a site where amino acid residues may be variable).
  • Such mutations are herein abbreviated as "D2G” mutations and may be resistant to enzymatic cleavage.
  • furin cleavage sites comprising RXXR sequences are mutated to AXXA.
  • AXXA sequences may also be resistant to enzymatic cleavage.
  • regions of proteolytic processing by tolloid and/or tolloid- like proteins may be mutated to prevent such proteolytic processing.
  • tolloid processing regions on GDF-8 and/or GDF-11 may be mutated.
  • mutation of aspartic acid residues to alanine residues within tolloid processing regions prevents tolloid processing. Mutation of aspartic acid residue 76 (D76) of the GDF-8 (myostatin) proprotein has been shown to prevent proteolytic activation of latent GDF-8 (Wolfman, N.M. et al.,PNAS. 2003, Oct 6. 100(26): 15842-6).
  • Asp 120 (D120, residue number counted from the translated protein, D98 from the proprotein of SEQ ID NO: 4) in GDF-11 may be mutated to prevent tolloid processing (Ge et al., 2005. Mol Cell Biol. 25(14):5846-58, the contents of which are herein incorporated by reference in their entirety).
  • one or more amino acids may be mutated in order to form recombinant GPCs with reduced latency. Such mutations are referred to herein as "activating mutations.” These mutations may introduce one or more regions of steric clash between complex prodomains and growth factor domains.
  • steric clash when referring to the interaction between two proteins or between two domains and/or epitopes within the same protein, refers to a repulsive interaction between such proteins, domains and/or epitopes due to overlapping position in three-dimensional space.
  • Steric clash within GPCs may reduce the affinity between prodomains and growth factor domains, resulting in elevated ratios of free growth factor to latent growth factor.
  • one or more amino acids may be mutated in order to form recombinant GPCs with increased latency. Such mutations are referred to herein as "stabilizing mutations.” These mutations may increase the affinity between prodomains and growth factor domains, resulting in decreased ratios of free growth factor to latent growth factor.
  • GDFs growth differentiation factors
  • GDFs Growth differentiation factors
  • activins and inhibins are TGF- ⁇ family member proteins involved in a number of cellular and/or developmental activities.
  • recombinant proteins may comprise one or more protein modules from one or more GDFs, activins and/or inhibins.
  • GDF protein modules may comprise GDF-8 and/or GDF-11 protein modules.
  • GDF-8 and GDF-11 which are secreted as latent complexes (Sengle et al., 2011. J Biol Chem. 286(7):5087-99; Ge et al., 2005. Mol Cel Biol. 25(14):5846-58), show
  • GDF-8 (also referred to herein as myostatin) is involved in regulating muscle mass, and its deficiency increases muscle mass in multiple species, including humans (Rodino-Klapac, L.R. et al., 2009. Muscle Nerve. 39(3):283-96). GDF-8 may be found in the circulation in latent form, but may also be stored in the extracellular matrix, bound to LTBP3 (Anderson et al., 2007. J Biol Chem. 283(11):7027-35) or perlecan (Sengle et al., 2011. J Biol Chem. 286(7):5087-99).
  • GDF-8 While complexed with its prodomain, GDF-8 is unable to participate in receptor binding with the type II receptor, ActRIIB (Sengle et al., 2008. J Mol Biol. 381(4): 1025-39). While GDF-8 is expressed primarily in muscle, GDF-11 expression is more systemic and its activity is thought to be involved in multiple processes (Lee et al., 2013. PNAS. 110(39):E3713-22). It is believed to be involved in development of multiple tissues, including, but not limited to the retina, kidney, pancreas and olfactory system. It is also believed to be a circulating factor in the blood. Recent studies indicate that GDF-11 may rejuvenate skeletal muscle, improve cerebral circulation and promote neurogenesis (Sinha, M. et al., 2014. Science.
  • antibodies of the disclosure may promote skeletal muscle rejuvenation, improve cerebral circulation and promote neurogenesis by promoting the release of GDF- 11 growth factor from latent complexes.
  • GDF-11 has also been identified as a regulator of erythropoiesis.
  • antibodies of the invation that inhibit GDF-11 may restore the ability of erythroid progenitors to differentiate and hence alleviate anemia (Paulson, RF. 2014. Nature Medicine. 20:334-5, the contents of which are herein incorporated by reference in their entirety).
  • GDF-8 and GDF-11 also share considerable homology. While the prodomains only share 48% homology, GDF-8 and GDF-11 growth factor domains share 90% homology (60% homology when prodomains and growth factors are taken together). Given the high degree of sequence similarity, it is not surprising that GDF- 11 and 8 growth factors bind and signal through the same receptors consisting of a Type I receptor (ALK4/5) in association with a type II receptor (ACTRIIA/B). The high degree of conservation in the mature growth factors has made it challenging to identify reagents and monoclonal antibodies that can differentiate between mature GDF-11 and 8. Consequently, there are no therapies in clinical trials today that are specific for GDF-11.
  • ALK4/5 Type I receptor
  • ACTRIIA/B type II receptor
  • proGDF proteins require furin cleavage in order to form latent complexes (see FIG. 9).
  • furin cleavage is carried out by cells in which proGDF proteins are expressed.
  • furin cleavage may be carried out through furin addition to cultures or solutions comprising proGDF proteins.
  • Furin cleavage of recombinant proteins of the disclosure may in some cases occur intracellularly. In some cases furin cleavage of recombinant proteins of the disclosure may occur extracellularly. Release of GDF-8 and GDF-11 from latent GPCs requires activating cleavage of latent complexes.
  • An activating cleavage refers to a proteolytic cleavage of a latent complex that leads to growth factor release and/or activation.
  • Activating cleavage may include prodomain cleavage at a BMP/tolloid/tolloid-like proteinase cleavage site.
  • sites may include the site located between Arg 75 and Asp 76 in GDF-8 and between Gly 97 and Asp 98 in GDF-11. This cleavage is between the 2 helix and the fastener.
  • recombinant proteins of the present disclosure comprising GDFs may comprise sequences listed in Table 4 or fragments thereof. In some cases, these sequences are expressed in association with N- and/or C-terminal secretion signal sequences [e.g. human Ig kappa chains with amino acid sequence MDMRVPAQLLGLLLLWFSGVLG (SEQ ID NO: 66)], flag tag sequences [e.g. DYKDDDDK (SEQ ID NO: 67)], one or more 3C protease cleavage site [e.g. LEVLFQGP (SEQ ID NO: 68)], one or more biotinylation site and/or His-tag sequences [e.g. HHHHHH (SEQ ID NO: 69)].
  • N- and/or C-terminal secretion signal sequences e.g. human Ig kappa chains with amino acid sequence MDMRVPAQLLGLLLLWFSGVLG (SEQ ID NO: 66)]
  • flag tag sequences e.
  • proGDF-8 AXXA NENSEQKENVEKEGLCNACTWRQNTKSSRIEAIKIQILSKLRLE 73 D76A TAPNISKDVIRQLLPKAPPLRELIDQYDVQRADSSDGSLEDDDY
  • proGDF-11 AEGPAAAAAAAAAAAAAGVGGERSSRPAPSVAPEPDGCPVC 4
  • proGDF-11 D98A AEGPAAAAAAAAAAAAAGVGGERSSRPAPSVAPEPDGCPVC 74
  • proGDF-11 AxxA AEGPAAAAAAAAAAAAAGVGGERSSRPAPSVAPEPDGCPVC 76
  • proGDF-11 AxxA AEGPAAAAAAAAAAAGVGGERSSRPAPSVAPEPDGCPVC 77 D98A VWRQHSRELRLESIKSQILSKLRLKEAPNISREVVKQLLPKAPP
  • Activins and inhibins are TGF- ⁇ family member proteins, the activity of each of which often results in opposing functions (Bilezikjian, L.M. et al., 2012. Mol Cell
  • Activins and inhibins are constructed in part from the same ⁇ - subunits, that may include inhibin-beta A, inhibin-beta B, inhibin-beta C and inhibin-beta E (referred to herein as ⁇ -subunit A, B, C and E, respectively).
  • ⁇ -subunit A, B, C and E The difference between activins and inhibins, structurally, is that activins are ⁇ -subunit dimers while inhibins are
  • Activins are named for their subunit pairs, such that activin A comprises a homodimer of two A subunits, activin AB comprises a dimer of A and B subunits, B comprises a dimer of B subunits, etc. (Muenster, U. et al., 2011. Vitam Horm. 85: 105-28). Activins are involved in a variety of functions that may include, but are not limited to cell growth, differentiation, programmed cell death, endocrine functions, cellular metabolism, bone growth, etc. They are especially recognized for their control of reproductive hormone cycles. Activin and inhibin signaling often functions antagonistically in this regard.
  • recombinant proteins of the present disclosure may comprise integrins.
  • Integrins are cell surface heterodimers formed by alpha and beta subunits, each of which has a transmembrane domain and in the N-terminal portion of the extracellular domain come together to form the ligand binding site.
  • Recombinant proteins of the present disclosure may comprise integrins and/or integrin subunits.
  • Such integrins and/or integrin subunits may comprise any of those disclosed in International Patent Application No.
  • Recombinant proteins of the disclosure may include intercellular adhesion molecule 1 (ICAM-1).
  • ICAM-1 proteins of the present disclosure may be used as control proteins during antibody development and/or antibody testing.
  • ICAM-1 may be used as a control during selection of binding molecules using phage display technologies.
  • ICAM-1 proteins of the disclosure comprise one or more detectable label. Detectable labels may include, for example, histidine tags.
  • recombinant GPCs of the present disclosure may comprise mutations in one or more N-terminal regions for extracellular associations.
  • N-terminal region for extracellular association refers to regions at or near protein N-termini that may be necessary for extracellular associations with one or more N-terminal regions. Such regions may comprise at least the first N-terminal residue, at least the first 5 N- terminal residues, at least the first 10 N-terminal residues, at least the first 20 amino acid residues and/or at least the first 50 amino acid residues.
  • Some mutations may comprise from about 1 amino acid residue to about 30 amino acid residues, from about 5 amino acid residues to about 40 amino acid residues and/or from about 10 amino acid residues to about 50 amino acid residues at or near protein N-termini.
  • Such regions may comprise residues for association with extracellular proteins (e.g. LTBPs, fibrillins, GASPs, perlecan and/or decorin).
  • LTBPs extracellular proteins
  • GASPs perlecan and/or decorin
  • cysteine residues present within and/or near N-terminal regions for extracellular associations may be necessary for such associations.
  • extracellular associations are present within about the first 5 N-terminal residues, about the first 10 N-terminal residues, about the first 20 N-terminal residues, about the first 50 N- terminal residues, about the first 60 N-terminal residues, about the first 70 N-terminal residues and/or at least the first 100 N-terminal residues.
  • Some mutations in one or more N- terminal regions for extracellular associations comprise substitution and/or deletion of such cysteine residues.
  • Such mutations may modulate the association of GPCs and/or prodomains with one or more extracellular proteins, including, but not limited to LTBPs, fibrillins, GASPs, perlecan and/or decorin. These mutations may also comprise substitution of one or more cysteine with another amino acid.
  • Cysteine residue substitutions are abbreviated herein as "C#X" wherein # represents the residue number [counting from the N-terminus of the pro- protein (without the signal peptide)] of the original cysteine residue and X represents the one letter amino acid code for the amino acid that is used for substitution. Any amino acid may be used for such substitutions.
  • serine (S) residues are used to substitute cysteine residues.
  • Nonlimiting examples of such mutations in GDF-8 may include C16S, C19S, C38S and/or C41S.
  • one or more cysteine in one or more other region of GPCs may be substituted or deleted.
  • such GPC modifications may promote the release of mature growth factor from prodomains.
  • cysteines or other residues may include those present in one or more of mature growth factors, alpha 2 helices, fasteners, latency lassos and/or bow-tie regions.
  • recombinant proteins of the present disclosure may comprise protein modules derived from one or more species, including mammals, including, but not limited to mice, rats, rabbits, pigs, monkeys and/or humans.
  • Recombinant proteins may comprise one or more amino acids from one or more amino acid sequences derived from one or more non-human protein sequences listed in Table 5.
  • recombinant proteins of the present disclosure may comprise such sequences with or without the native signal peptide. In some cases, these sequences are expressed in association with N- and/or C- terminal secretion signal sequences [e.g. human Ig kappa chains with amino acid sequence MDMRVPAQLLGLLLLWFSGVLG (SEQ ID NO: 66)], flag tag sequences [e.g.
  • DYKDDDDK (SEQ ID NO: 67)]
  • one or more 3C protease cleavage site e.g. LEVLFQGP (SEQ ID NO: 68)
  • one or more biotinylation site and/or His-tag sequences e.g. HHHHHH (SEQ ID NO: 69)].
  • proGDF-8 Mouse NEGSEREENVEKEGLCNACAWRQNTRYSRIEAIKIQILS 88
  • proGDF-8 Mouse NEGSEREENVEKEGLCNACAWRQNTRYSRIEAIKIQILS 89 AxxA KLRLETAPNISKDAIRQLLPRAPPLRELIDQYDVQRDDSS
  • proGDF-8 Mouse NEGSEREENVEKEGLCNACAWRQNTRYSRIEAIKIQILS 90 D76A KLRLETAPNISKDAIRQLLPRAPPLRELIDQYDVQRADSS
  • proGDF-8 Mouse NEGSEREENVEKEGLCNACAWRQNTRYSRIEAIKIQILS 91 AxxA D76A KLRLETAPNISKDAIRQLLPRAPPLRELIDQYDVQRADSS

Abstract

L'invention concerne des protéines, des anticorps, des essais, des compositions et des méthodes servant à moduler les taux et/ou les activités de facteurs de croissance. Dans certains modes de réalisation, ces facteurs de croissance font partie de la superfamille de protéines TGF-β.
PCT/US2015/059557 2014-11-06 2015-11-06 Immunoessais liés à des facteurs de croissance transformants WO2016073906A2 (fr)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US201462076230P 2014-11-06 2014-11-06
US62/076,230 2014-11-06
US201562100361P 2015-01-06 2015-01-06
US62/100,361 2015-01-06
US201562187348P 2015-07-01 2015-07-01
US62/187,348 2015-07-01

Publications (2)

Publication Number Publication Date
WO2016073906A2 true WO2016073906A2 (fr) 2016-05-12
WO2016073906A3 WO2016073906A3 (fr) 2016-07-21

Family

ID=55910042

Family Applications (2)

Application Number Title Priority Date Filing Date
PCT/US2015/059515 WO2016073879A2 (fr) 2014-11-06 2015-11-06 Anticorps liés à des facteurs de croissance transformants et utilisations de ceux-ci
PCT/US2015/059557 WO2016073906A2 (fr) 2014-11-06 2015-11-06 Immunoessais liés à des facteurs de croissance transformants

Family Applications Before (1)

Application Number Title Priority Date Filing Date
PCT/US2015/059515 WO2016073879A2 (fr) 2014-11-06 2015-11-06 Anticorps liés à des facteurs de croissance transformants et utilisations de ceux-ci

Country Status (1)

Country Link
WO (2) WO2016073879A2 (fr)

Cited By (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9828429B2 (en) 2007-09-26 2017-11-28 Chugai Seiyaku Kabushiki Kaisha Method of modifying isoelectric point of antibody via amino acid substitution in CDR
US9868948B2 (en) 2008-04-11 2018-01-16 Chugai Seiyaku Kabushiki Kaisha Antigen-binding molecule capable of binding to two or more antigen molecules repeatedly
US9969800B2 (en) 2015-02-05 2018-05-15 Chugai Seiyaku Kabushiki Kaisha IL-8 antibodies
US10000560B2 (en) 2014-12-19 2018-06-19 Chugai Seiyaku Kabushiki Kaisha Anti-myostatin antibodies, polypeptides containing variant Fc regions, and methods of use
WO2018132572A1 (fr) * 2017-01-11 2018-07-19 The Board Of Trustees Of The Leland Stanford Junior University Molécules de substitution de r-spondine (rspo)
WO2019055399A1 (fr) * 2017-09-12 2019-03-21 Igc Bio, Inc. Anticorps anti-vegf
US10307480B2 (en) 2014-11-06 2019-06-04 Scholar Rock, Inc. Anti-pro/latent-myostatin antibodies and uses thereof
US10751413B2 (en) 2015-09-15 2020-08-25 Scholar Rock, Inc. Anti-pro/latent-Myostatin antibodies and uses thereof
US10882904B2 (en) 2016-01-08 2021-01-05 Scholar Rock, Inc. Methods for inhibiting myostatin activation by administering anti-pro/latent myostatin antibodies
US10919953B2 (en) 2012-08-24 2021-02-16 Chugai Seiyaku Kabushiki Kaisha FcgammaRIIB-specific Fc region variant
US10946036B2 (en) 2016-06-13 2021-03-16 Scholar Rock, Inc. Use of myostatin inhibitors and combination therapies
US11046784B2 (en) 2006-03-31 2021-06-29 Chugai Seiyaku Kabushiki Kaisha Methods for controlling blood pharmacokinetics of antibodies
US11046777B2 (en) * 2016-07-22 2021-06-29 Dana-Farber Cancer Institute, Inc. Glucocorticoid-induced tumor necrosis factor receptor (GITR) antibodies and methods of use thereof
US11053308B2 (en) 2016-08-05 2021-07-06 Chugai Seiyaku Kabushiki Kaisha Method for treating IL-8-related diseases
US11155611B2 (en) 2017-01-06 2021-10-26 Scholar Rock, Inc. Compositions and methods for making and using anti-myostatin antibodies
US11168128B2 (en) 2020-02-26 2021-11-09 Vir Biotechnology, Inc. Antibodies against SARS-CoV-2 and methods of using the same
US11236168B2 (en) 2012-08-24 2022-02-01 Chugai Seiyaku Kabushiki Kaisha Mouse FcγammaRII-specific Fc antibody
US11267868B2 (en) 2013-04-02 2022-03-08 Chugai Seiyaku Kabushiki Kaisha Fc region variant
US11359009B2 (en) 2015-12-25 2022-06-14 Chugai Seiyaku Kabushiki Kaisha Anti-myostatin antibodies and methods of use
WO2022125482A1 (fr) * 2020-12-08 2022-06-16 Memorial Sloan Kettering Cancer Center Anticorps dirigés contre la galectine-3 et leurs méthodes d'utilisation
US11401339B2 (en) 2018-08-23 2022-08-02 Seagen Inc. Anti-TIGIT antibodies
US11427632B2 (en) 2016-07-06 2022-08-30 Celgene Corporation Antibodies with low immunogenicity and uses thereof
WO2023037119A1 (fr) * 2021-09-10 2023-03-16 Samatva Research Corporation Fraction anti-virus immobilisée dans une matrice
US11891434B2 (en) 2010-11-30 2024-02-06 Chugai Seiyaku Kabushiki Kaisha Antigen-binding molecule capable of binding to plurality of antigen molecules repeatedly
US11958891B2 (en) 2017-01-26 2024-04-16 Surrozen Operating, Inc. Tissue-specific Wnt signal enhancing molecules and uses thereof

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3324996A4 (fr) * 2015-07-22 2019-04-17 Scholar Rock, Inc. Protéines de liaison à gdf11 et leurs utilisations
AU2016372934B2 (en) * 2015-12-18 2023-10-05 Chugai Seiyaku Kabushiki Kaisha Anti-myostatin antibodies, polypeptides containing variant Fc regions, and methods of use
KR102456739B1 (ko) * 2016-06-17 2022-10-19 추가이 세이야쿠 가부시키가이샤 항-마이오스타틴 항체 및 사용 방법
KR20190049866A (ko) * 2016-09-20 2019-05-09 우시 바이올로직스 아일랜드 리미티드 신규한 항-ｐｃｓｋ９ 항체
TWI782920B (zh) * 2016-09-20 2022-11-11 中國大陸商上海藥明生物技術有限公司 新型抗-pcsk9抗體
RU2770590C2 (ru) 2016-10-30 2022-04-18 Шанхай Хенлиус Байотек, Инк. Антитела против pd-l1 и их варианты
WO2018129395A1 (fr) * 2017-01-06 2018-07-12 Scholar Rock, Inc. Méthodes de traitement de maladies métaboliques par inhibition de l'activation de la myostatine
CN113429479B (zh) * 2018-04-04 2023-03-10 中国科学院微生物研究所 一种高灵敏度的黄热病毒人源单克隆抗体及其应用
WO2019210144A1 (fr) * 2018-04-27 2019-10-31 Vanderbilt University Anticorps largement neutralisants dirigés contre le virus de l'hépatite c
CA3105454A1 (fr) * 2018-07-03 2020-01-09 Catalent Pharma Solutions, Llc Molecules proteiques multifonctionnelles comprenant de la decorine et utilisation associee
WO2020264410A1 (fr) * 2019-06-26 2020-12-30 Sensor-Kinesis Corporation Agents de liaison à la troponine t et utilisations de ceux-ci
CN111662379B (zh) * 2020-05-09 2021-03-02 江苏省疾病预防控制中心(江苏省公共卫生研究院) 抗新型冠状病毒的抗体、制备方法和应用
WO2023009891A2 (fr) * 2021-07-30 2023-02-02 Janssen Biotech, Inc. Matériaux et procédés de fabrication ou d'utilisation de protéines de liaison à il-23 r
CN115322257B (zh) * 2021-08-16 2023-05-30 上海优替济生生物医药有限公司 Bcma靶向抗体、嵌合抗原受体及其应用
WO2023086336A2 (fr) * 2021-11-09 2023-05-19 University Of Pittsburgh – Of The Commonwealth System Of Higher Education Molécules qui se lient à des polypeptides d'antigène membranaire spécifique de la prostate (psma)

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7393682B1 (en) * 1993-03-19 2008-07-01 The Johns Hopkins University School Of Medicine Polynucleotides encoding promyostatin polypeptides
DE69432815T2 (de) * 1993-03-19 2003-12-11 Univ Johns Hopkins Med Wachstumsfaktor-8
UA92504C2 (en) * 2005-10-12 2010-11-10 Эли Лилли Энд Компани Anti-myostatin monoclonal antibody
PE20091163A1 (es) * 2007-11-01 2009-08-09 Wyeth Corp Anticuerpos para gdf8
AU2011292197B2 (en) * 2010-08-16 2015-05-28 Amgen Inc. Antibodies that bind myostatin, compositions and methods
SG10201913751RA (en) * 2013-05-06 2020-03-30 Scholar Rock Inc Compositions and methods for growth factor modulation

Cited By (39)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11046784B2 (en) 2006-03-31 2021-06-29 Chugai Seiyaku Kabushiki Kaisha Methods for controlling blood pharmacokinetics of antibodies
US9828429B2 (en) 2007-09-26 2017-11-28 Chugai Seiyaku Kabushiki Kaisha Method of modifying isoelectric point of antibody via amino acid substitution in CDR
US11248053B2 (en) 2007-09-26 2022-02-15 Chugai Seiyaku Kabushiki Kaisha Method of modifying isoelectric point of antibody via amino acid substitution in CDR
US11359194B2 (en) 2008-04-11 2022-06-14 Chugai Seiyaku Kabushiki Kaisha Antigen-binding molecule capable of binding two or more antigen molecules repeatedly
US9868948B2 (en) 2008-04-11 2018-01-16 Chugai Seiyaku Kabushiki Kaisha Antigen-binding molecule capable of binding to two or more antigen molecules repeatedly
US10472623B2 (en) 2008-04-11 2019-11-12 Chugai Seiyaku Kabushiki Kaisha Antigen-binding molecule capable of binding two or more antigen molecules repeatedly
US9890377B2 (en) 2008-04-11 2018-02-13 Chugai Seiyaku Kabushiki Kaisha Antigen-binding molecule capable of binding to two or more antigen molecules repeatedly
US11371039B2 (en) 2008-04-11 2022-06-28 Chugai Seiyaku Kabushiki Kaisha Antigen-binding molecule capable of binding to two or more antigen molecules repeatedly
US11891434B2 (en) 2010-11-30 2024-02-06 Chugai Seiyaku Kabushiki Kaisha Antigen-binding molecule capable of binding to plurality of antigen molecules repeatedly
US10919953B2 (en) 2012-08-24 2021-02-16 Chugai Seiyaku Kabushiki Kaisha FcgammaRIIB-specific Fc region variant
US11236168B2 (en) 2012-08-24 2022-02-01 Chugai Seiyaku Kabushiki Kaisha Mouse FcγammaRII-specific Fc antibody
US11267868B2 (en) 2013-04-02 2022-03-08 Chugai Seiyaku Kabushiki Kaisha Fc region variant
US10307480B2 (en) 2014-11-06 2019-06-04 Scholar Rock, Inc. Anti-pro/latent-myostatin antibodies and uses thereof
US11135291B2 (en) 2014-11-06 2021-10-05 Scholar Rock, Inc. Methods for making and using anti-myostatin antibodies
US11925683B2 (en) 2014-11-06 2024-03-12 Scholar Rock, Inc. Compositions for making and using anti-Myostatin antibodies
US10738111B2 (en) 2014-12-19 2020-08-11 Chugai Seiyaku Kabushiki Kaisha Anti-myostatin antibodies, polypeptides containing variant Fc regions, and methods of use
US10000560B2 (en) 2014-12-19 2018-06-19 Chugai Seiyaku Kabushiki Kaisha Anti-myostatin antibodies, polypeptides containing variant Fc regions, and methods of use
US11454633B2 (en) 2014-12-19 2022-09-27 Chugai Seiyaku Kabushiki Kaisha Anti-myostatin antibodies, polypeptides containing variant Fc regions, and methods of use
US10519229B2 (en) 2015-02-05 2019-12-31 Chugai Seiyaku Kabushiki Kaisha Nucleic acids encoding IL-8 antibodies
US9969800B2 (en) 2015-02-05 2018-05-15 Chugai Seiyaku Kabushiki Kaisha IL-8 antibodies
US11180548B2 (en) 2015-02-05 2021-11-23 Chugai Seiyaku Kabushiki Kaisha Methods of neutralizing IL-8 biological activity
US11439704B2 (en) 2015-09-15 2022-09-13 Scholar Rock, Inc. Methods for using anti-myostatin antibodies
US10751413B2 (en) 2015-09-15 2020-08-25 Scholar Rock, Inc. Anti-pro/latent-Myostatin antibodies and uses thereof
US11359009B2 (en) 2015-12-25 2022-06-14 Chugai Seiyaku Kabushiki Kaisha Anti-myostatin antibodies and methods of use
US10882904B2 (en) 2016-01-08 2021-01-05 Scholar Rock, Inc. Methods for inhibiting myostatin activation by administering anti-pro/latent myostatin antibodies
US10946036B2 (en) 2016-06-13 2021-03-16 Scholar Rock, Inc. Use of myostatin inhibitors and combination therapies
US11427632B2 (en) 2016-07-06 2022-08-30 Celgene Corporation Antibodies with low immunogenicity and uses thereof
US11046777B2 (en) * 2016-07-22 2021-06-29 Dana-Farber Cancer Institute, Inc. Glucocorticoid-induced tumor necrosis factor receptor (GITR) antibodies and methods of use thereof
US11780912B2 (en) 2016-08-05 2023-10-10 Chugai Seiyaku Kabushiki Kaisha Composition for prophylaxis or treatment of IL-8 related diseases
US11053308B2 (en) 2016-08-05 2021-07-06 Chugai Seiyaku Kabushiki Kaisha Method for treating IL-8-related diseases
US11155611B2 (en) 2017-01-06 2021-10-26 Scholar Rock, Inc. Compositions and methods for making and using anti-myostatin antibodies
WO2018132572A1 (fr) * 2017-01-11 2018-07-19 The Board Of Trustees Of The Leland Stanford Junior University Molécules de substitution de r-spondine (rspo)
US11958891B2 (en) 2017-01-26 2024-04-16 Surrozen Operating, Inc. Tissue-specific Wnt signal enhancing molecules and uses thereof
WO2019055399A1 (fr) * 2017-09-12 2019-03-21 Igc Bio, Inc. Anticorps anti-vegf
US11401339B2 (en) 2018-08-23 2022-08-02 Seagen Inc. Anti-TIGIT antibodies
US11479599B2 (en) 2020-02-26 2022-10-25 Vir Biotechnology, Inc. Antibodies against SARS-CoV-2 and methods of using the same
US11168128B2 (en) 2020-02-26 2021-11-09 Vir Biotechnology, Inc. Antibodies against SARS-CoV-2 and methods of using the same
WO2022125482A1 (fr) * 2020-12-08 2022-06-16 Memorial Sloan Kettering Cancer Center Anticorps dirigés contre la galectine-3 et leurs méthodes d'utilisation
WO2023037119A1 (fr) * 2021-09-10 2023-03-16 Samatva Research Corporation Fraction anti-virus immobilisée dans une matrice

Also Published As

Publication number Publication date
WO2016073906A3 (fr) 2016-07-21
WO2016073879A2 (fr) 2016-05-12
WO2016073879A3 (fr) 2016-06-23

Similar Documents

Publication Publication Date Title
US11827698B2 (en) Compositions and methods for growth factor modulation
WO2016073906A2 (fr) Immunoessais liés à des facteurs de croissance transformants
US20170073406A1 (en) Compositions and methods for growth factor modulation
US20180208648A1 (en) Gdf11 binding proteins and uses thereof
US9783610B2 (en) Anti-tumor endothelial marker-1 (TEM1) antibody variants and uses thereof

Legal Events

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

Ref document number: 15857747

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase in:

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 15857747

Country of ref document: EP

Kind code of ref document: A2