Compositions and methods for growth factor modulation
First Claim
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1. A method of producing an antibody capable of binding a growth factor prodomain complex (GPC), said GPC consisting of at least one GDF-8 growth factor and at least one GDF-8 prodomain, said method comprising:
- a. expressing proGDF-8 (SEQ ID NO;
5),b. forming the GPC by subjecting the expressed proGDF-8 (SEQ ID NO;
5) 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),c. 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;
d. selecting phage particles bound to the GPC formed by enzymatic cleavage; and
e. producing recombinant antibodies having complementarity determining region (CDR) amino acid sequences obtained from the antibody fragments expressed at the surface of the phage particles selected in (d).
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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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7 Claims
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1. A method of producing an antibody capable of binding a growth factor prodomain complex (GPC), said GPC consisting of at least one GDF-8 growth factor and at least one GDF-8 prodomain, said method comprising:
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a. expressing proGDF-8 (SEQ ID NO;
5),b. forming the GPC by subjecting the expressed proGDF-8 (SEQ ID NO;
5) 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),c. 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; d. selecting phage particles bound to the GPC formed by enzymatic cleavage; and e. producing recombinant antibodies having complementarity determining region (CDR) amino acid sequences obtained from the antibody fragments expressed at the surface of the phage particles selected in (d). - View Dependent Claims (2, 3, 4, 5, 6, 7)
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Specification