Gpcr binding proteins and synthesis thereof

Abstract

Provided herein are methods and compositions relating to G protein-coupled receptor (GPCR) libraries having nucleic acids encoding for a scaffold comprising a GPCR binding domain. Libraries described herein include variegated libraries comprising nucleic acids each encoding for a predetermined variant of at least one predetermined reference nucleic acid sequence. Further described herein are protein libraries generated when the nucleic acid libraries are translated. Further described herein are cell libraries expressing variegated nucleic acid libraries described herein.

What is claimed is: 
     
         1 . An antibody, wherein the antibody comprises a CDR-H 3  comprising a sequence of any one of SEQ ID NOS: 2420 to 2436. 
     
     
         2 . The antibody of  claim 1 , wherein the antibody comprises a CDR-H 3  comprising a sequence of any one of SEQ ID NOS: 2420 to 2436; and wherein the antibody is a monoclonal antibody, a polyclonal antibody, a bi-specific antibody, a multispecific antibody, a grafted antibody, a human antibody, a humanized antibody, a synthetic antibody, a chimeric antibody, a camelized antibody, a single-chain Fvs (scFv), a single chain antibody, a Fab fragment, a F(ab′)2 fragment, a Fd fragment, a Fv fragment, a single-domain antibody, an isolated complementarity determining region (CDR), a diabody, a fragment comprised of only a single monomeric variable domain, disulfide-linked Fvs (sdFv), an intrabody, an anti-idiotypic (anti-Id) antibody, or ab antigen-binding fragments thereof. 
     
     
         3 . A method for treatment of a metabolic disorder, comprising administering to a subject in need thereof the antibody of  claim 1 . 
     
     
         4 . The method of  claim 3 , wherein the metabolic disorder is Type II diabetes or obesity. 
     
     
         5 . A protein library comprising a plurality of proteins, wherein each of the proteins of the plurality of proteins comprise an immunoglobulin scaffold, wherein the immunoglobulin scaffold comprises a CDR-H 3  loop that comprises a sequence variant of a GPCR binding domain. 
     
     
         6 . A protein library comprising a plurality of proteins, wherein the plurality of proteins comprises sequence encoding for different GPCR binding domains, and wherein the length of each GPCR binding domain is about 20 to about 80 amino acids. 
     
     
         7 . An antibody library comprising a plurality of antibodies, wherein each antibody comprises:
 a. a CDR-H 3  loop comprising an amino acid sequence having at least about 90% sequence identity to any one of SEQ ID NOs: 2420 to 2436; and   b. a variable domain of light chain (VL) comprising an amino acid sequence having at least about 90% sequence identity to any one of SEQ ID NOs: 91, 93, 95, 97, 100, 118, 138, 233, 249, 276, 277, 465, 467, 496, 501, 509, 515, 526, 594, 604, 803, 1059, and 1219,   wherein each of the antibodies comprises a GPCR binding domain, and wherein the GPCR binding domain is a ligand for a GPCR.   
     
     
         8 . The antibody library of  claim 7 , wherein the antibody further comprises one or more domains selected from variable domain of heavy chain (VH), constant domain of light chain (CL), and constant domain of heavy chain (CH). 
     
     
         9 . The antibody library of  claim 7 , wherein the GPCR binding domains comprise peptidomimetic or small molecule mimetic. 
     
     
         10 . The antibody library of  claim 7 , wherein the ligand for a GPCR is a non-antibody ligand. 
     
     
         11 . The antibody library of  claim 7 , wherein the CDR-H 3  loop comprises an amino acid sequence of any one of SEQ ID NOs: 2420 to 2436. 
     
     
         12 . The antibody library of  claim 7 , wherein the variable domain of light chain (VL) comprises an amino acid sequence of any one of SEQ ID NOs: 91, 93, 95, 97, 100, 118, 138, 233, 249, 276, 277, 465, 467, 496, 501, 509, 515, 526, 594, 604, 803, 1059, and 1219. 
     
     
         13 . The antibody library of  claim 7 , wherein the variable domain of light chain (VL) comprises an amino acid sequence having at least about 90% sequence identity to any one of SEQ ID NOs: 91, 93, 95, and 97. 
     
     
         14 . The antibody library of  claim 7 , wherein the variable domain of light chain (VL) comprises an amino acid sequence of any one of SEQ ID NOs: 91, 93, 95, and 97. 
     
     
         15 . The antibody library of  claim 7 , wherein the variable domain of light chain (VL) comprises an amino acid sequence having at least about 90% sequence identity to any one of SEQ ID NOs: 276, 604, 803, 1059, and 1219. 
     
     
         16 . The antibody library of  claim 7 , wherein the variable domain of light chain (VL) comprises an amino acid sequence of any one of SEQ ID NOs: 276, 604, 803, 1059, and 1219. 
     
     
         17 . The antibody library of  claim 7 , wherein the antibody is a monoclonal antibody, a polyclonal antibody, a bi-specific antibody, a multispecific antibody, a grafted antibody, a human antibody, a humanized antibody, a synthetic antibody, a chimeric antibody, a camelized antibody, a single-chain Fvs (scFv), a single chain antibody, a Fab fragment, a F(ab′)2 fragment, a Fd fragment, a Fv fragment, a single-domain antibody, an isolated complementarity determining region (CDR), a diabody, a fragment comprised of only a single monomeric variable domain, disulfide-linked Fvs (sdFv), an intrabody, an anti-idiotypic (anti-Id) antibody, or ab antigen-binding fragments thereof. 
     
     
         18 . A method of inhibiting GLP1R activity, comprising administering the antibody library of  claim 7 . 
     
     
         19 . A method for treatment of a metabolic disorder, comprising administering the antibody library of  claim 7 .