Rational Design of a Humanized Glucagon-Like Peptide-1 Receptor Agonist Antibody

• Published in: Angewandte Chemie Vol. 127; no. 7; pp. 2154 - 2158
• Main Authors: Zhang, Yong, Zou, Huafei, Wang, Ying, Caballero, Dawna, Gonzalez, Jose, Chao, Elizabeth, Welzel, Gus, Shen, Weijun, Wang, Danling, Schultz, Peter G., Wang, Feng
• Format: Journal Article
• Language: English; German
• Published: Weinheim WILEY-VCH Verlag 09.02.2015; WILEY‐VCH Verlag; Wiley Subscription Services, Inc
• Subjects: Antibodies; Antikörper; Blood; Chemistry; Cleavage; Coiling; Glucose; Peptide; Peptides; Pharmakologie; Protein-Engineering; Proteins; Receptors; Rezeptoren
• ISSN: 0044-8249; 1521-3757
• DOI: 10.1002/ange.201410049

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. Chimärenbildung: Ein Antikörper gegen den GLP‐1‐Rezeptoragonisten (GLP‐1=glucagon‐like peptide‐1) wurde durch genetische Fusion von Exendin‐4 mit der CDR3H‐Region des humanisierten monoklonalen Antikörpers Herceptin mit einem heterodimeren Coiled‐Coil‐„Stiel“ erzeugt. Das resultierende gekürzte Herceptin‐Ex‐4‐Fusionsprotein zeichnet sich durch gute biologische und pharmakologische Eigenschaften aus.