Compositions and methods for growth factor modulation
First Claim
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1. An antibody produced by a method comprising the steps of:
- i. expressing mammalian proGDF-8;
ii. subjecting the expressed mammalian proGDF-8 to enzymatic cleavage with one or more of furin, bone morphogenetic protein-1 (BMP-1), mammalian tolloid protein (mTLD), mammalian tolloid-like 1 (mTLL1), and mammalian tolloid-like 2 (mTLL2), so as to form a growth factor prodomain complex (GPC);
iii. carrying out solid-phase or solution-phase enrichment with an antibody fragment phage display library, wherein the GPC formed by enzymatic cleavage is used as a target antigen;
iv. selecting phage particles that bind to the GPC formed by enzymatic cleavage and excluding phage particles that bind to isolated GDF-8 growth factor or isolated GDF-8 prodomain; and
v. producing recombinant antibodies having complementarity determining regions (CDRs) of an antibody fragment expressed at the surface of the selected phage particles.
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Abstract
Provided herein are proteins, antibodies, assays and methods useful for modulating growth factor levels and/or activities. In some embodiments, such growth factors are members of the TGF-β superfamily of proteins.
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Citations
16 Claims
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1. An antibody produced by a method comprising the steps of:
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i. expressing mammalian proGDF-8; ii. subjecting the expressed mammalian proGDF-8 to enzymatic cleavage with one or more of furin, bone morphogenetic protein-1 (BMP-1), mammalian tolloid protein (mTLD), mammalian tolloid-like 1 (mTLL1), and mammalian tolloid-like 2 (mTLL2), so as to form a growth factor prodomain complex (GPC); iii. carrying out solid-phase or solution-phase enrichment with an antibody fragment phage display library, wherein the GPC formed by enzymatic cleavage is used as a target antigen; iv. selecting phage particles that bind to the GPC formed by enzymatic cleavage and excluding phage particles that bind to isolated GDF-8 growth factor or isolated GDF-8 prodomain; and v. producing recombinant antibodies having complementarity determining regions (CDRs) of an antibody fragment expressed at the surface of the selected phage particles. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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Specification