title: The dimerization of glucagon-like peptide-2 MIMETIBODY™ is linked to leucine-17 in the glucagon-like peptide-2 region.
abstract: Glucagon-like peptide-2 (GLP-2) is a member of the glucagon multigene family that is produced by intestinal enteroendocrine cells in response to food intake. GLP-2 stimulates growth of the intestinal epithelium, enhances its barrier functions, and increases nutrient uptake. Therefore, a GLP-2 agonist may be efficacious in human diseases characterized by malabsorption or injury to the gastrointestinal epithelium. MIMETIBODY™ refers to a proprietary scaffold developed to extend the half-life of rapidly cleared peptides. It consists of a peptide linked to a scaffold that contains sequence elements from a human immunoglobulin G including those that allow recycling through the FcRn. The GLP-2 sequence was engineered into the MIMETIBODY™ scaffold. The primary state of both GLP-2 and the GLP-2 MIMETIBODY™ in DPBS was a noncovalently associated dimer indicative of self-interaction. The increased heterogeneity and the decreased lot-to-lot reproducibility caused by the self-interaction of therapeutic proteins are a challenge to drug development. A similar protein, GLP-1 MIMETIBODY™, contains the related GLP-1 peptide and does not form a dimer under similar conditions. Therefore, to minimize or abrogate dimerization, several variants were made by substituting GLP-2 amino acids with the corresponding amino acids from GLP-1. Molecular weight and secondary structure analyses reveal that substituting leucine for glutamine at position 17 (L17Q) reduces dimerization and α-helix content yet retains bioactivity.
authors.affiliation: Biologics Research Department, Centocor R&D, Radnor, PA, USA. abaker8@its.jnj.com
authors.forename: Audrey E
authors.initials: AE
authors.lastname: Baker
authors.forename: Sarah
authors.initials: S
authors.lastname: Sague
authors.forename: Tami L R
authors.initials: TL
authors.lastname: Grygiel
authors.forename: Albert
authors.initials: A
authors.lastname: Schmidt
authors.forename: Alison
authors.initials: A
authors.lastname: Rogers
authors.forename: Haiyan
authors.initials: H
authors.lastname: Jiang
authors.forename: Marian
authors.initials: M
authors.lastname: Kruszynski
authors.forename: Tom
authors.initials: T
authors.lastname: Nesspor
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D002850:Chromatography, Gel; D000242:Cyclic AMP / Q000096:biosynthesis; D052216:Glucagon-Like Peptide 1 / Q000737:chemistry; D053767:Glucagon-Like Peptide 2 / Q000737:chemistry* / Q000235:genetics; D000067757:Glucagon-Like Peptide-1 Receptor; D000067758:Glucagon-Like Peptide-2 Receptor; D057809:HEK293 Cells; D006801:Humans; D007930:Leucine / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D018027:Receptors, Glucagon / Q000819:agonists / Q000378:metabolism
chemical_list: C000593833:GLP1R protein, human; D053767:Glucagon-Like Peptide 2; D000067757:Glucagon-Like Peptide-1 Receptor; D000067758:Glucagon-Like Peptide-2 Receptor; D010446:Peptide Fragments; D018027:Receptors, Glucagon; D052216:Glucagon-Like Peptide 1; D000242:Cyclic AMP; D007930:Leucine