Rational design of a humanized glucagon-like peptide-1 receptor agonist antibody

Angew Chem Int Ed Engl. 2015 Feb 9;54(7):2126-30. doi: 10.1002/anie.201410049. Epub 2014 Dec 29.

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.

Keywords: antibodies; peptide; pharmacology; protein engineering; receptors.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal, Humanized / chemistry
  • Antibodies, Monoclonal, Humanized / genetics
  • Antibodies, Monoclonal, Humanized / immunology*
  • Antibodies, Monoclonal, Humanized / pharmacology
  • Cattle
  • Exenatide
  • Glucagon-Like Peptide-1 Receptor
  • HEK293 Cells
  • Humans
  • Mice
  • Models, Molecular
  • Molecular Sequence Data
  • Peptides / chemistry
  • Peptides / genetics
  • Peptides / immunology*
  • Peptides / pharmacology
  • Protein Engineering
  • Receptors, Glucagon / agonists*
  • Receptors, Glucagon / immunology*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology*
  • Recombinant Fusion Proteins / pharmacology
  • Trastuzumab
  • Venoms / chemistry
  • Venoms / genetics
  • Venoms / immunology*
  • Venoms / pharmacology

Substances

  • Antibodies, Monoclonal, Humanized
  • GLP1R protein, human
  • Glp1r protein, mouse
  • Glucagon-Like Peptide-1 Receptor
  • Peptides
  • Receptors, Glucagon
  • Recombinant Fusion Proteins
  • Venoms
  • Exenatide
  • Trastuzumab