title: Rational design of a humanized glucagon-like peptide-1 receptor agonist antibody.
abstract: Bovine antibody BLV1H12 possesses a unique "stalk-knob" architecture in its ultralong heavy chain CDR3, allowing substitutions of the "knob" domain with protein agonists to generate functional antibody chimeras. We have generated a humanized glucagon-like peptide-1 (GLP-1) receptor agonist antibody by first introducing a coiled-coil "stalk" into CDR3H of the antibody herceptin. Exendin-4 (Ex-4), a GLP-1 receptor agonist, was then fused to the engineered stalk with flexible linkers, and a Factor Xa cleavage site was inserted immediately in front of Ex-4 to allow release of the N-terminus of the fused peptide. The resulting clipped herceptin-Ex-4 fusion protein is more potent in vitro in activating GLP-1 receptors than the Ex-4 peptide. The clipped herceptin-Ex-4 has an extended plasma half-life of approximately four days and sustained control of blood glucose levels for more than a week in mice. This work provides a novel approach to the development of human or humanized agonist antibodies as therapeutics.
authors.affiliation: California Institute for Biomedical Research (Calibr), 11119 N. Torrey Pines Road, La Jolla, CA 92037 (USA).
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Huafei
authors.initials: H
authors.lastname: Zou
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Dawna
authors.initials: D
authors.lastname: Caballero
authors.forename: Jose
authors.initials: J
authors.lastname: Gonzalez
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Chao
authors.forename: Gus
authors.initials: G
authors.lastname: Welzel
authors.forename: Weijun
authors.initials: W
authors.lastname: Shen
authors.forename: Danling
authors.initials: D
authors.lastname: Wang
authors.forename: Peter G
authors.initials: PG
authors.lastname: Schultz
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D002417:Cattle; D000077270:Exenatide; D000067757:Glucagon-Like Peptide-1 Receptor; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D015202:Protein Engineering; D018027:Receptors, Glucagon / Q000819:agonists* / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D000068878:Trastuzumab; D014688:Venoms / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology
chemical_list: D061067:Antibodies, Monoclonal, Humanized; C000593833:GLP1R protein, human; C000593834:Glp1r protein, mouse; D000067757:Glucagon-Like Peptide-1 Receptor; D010455:Peptides; D018027:Receptors, Glucagon; D011993:Recombinant Fusion Proteins; D014688:Venoms; D000077270:Exenatide; D000068878:Trastuzumab
keywords: antibodies; peptide; pharmacology; protein engineering; receptors