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Rational Design of a Humanized Glucagon‐Like Peptide‐1 Receptor Agonist Antibody
Author(s) -
Zhang Yong,
Zou Huafei,
Wang Ying,
Caballero Dawna,
Gonzalez Jose,
Chao Elizabeth,
Welzel Gus,
Shen Weijun,
Wang Danling,
Schultz Peter G.,
Wang Feng
Publication year - 2015
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201410049
Subject(s) - agonist , receptor , antibody , chemistry , peptide , ex vivo , fusion protein , microbiology and biotechnology , biochemistry , recombinant dna , in vitro , biology , immunology , gene
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.