Heterogeneity of recombinant antibodies: linking structure to function

Dev Biol (Basel). 2005;122:117-27.

Abstract

Structural heterogeneity of recombinant IgG1 antibodies derives from variations in conserved as well as unique structural features. Common sources of heterogeneity include Fc glycosylation, partial heavy chain C-terminal Lys processing, Fc methionine oxidation, hinge-region cleavage, and the glycation of Lys residues. Aspartate residues that are isomerized to iso-aspartate were detected by cation exchange or hydrophobic interaction chromatography for trastuzumab and omalizumab, respectively. Unpaired cysteines were detected in omalizumab using Ellman's reagent, with the thiol-containing Fab resolved using hydrophobic interaction chromatography after papain digestion. Structural variations that cause chromatographic resolution may indicate the presence of a form with reduced potency.

MeSH terms

  • Animals
  • Antibodies, Anti-Idiotypic
  • Antibodies, Monoclonal / analysis*
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal, Humanized
  • Antineoplastic Agents / analysis*
  • Antineoplastic Agents / chemistry
  • Humans
  • Immunoglobulin G / analysis*
  • Immunoglobulin G / chemistry
  • Omalizumab
  • Protein Modification, Translational*
  • Protein Processing, Post-Translational*
  • Structure-Activity Relationship
  • Trastuzumab

Substances

  • Antibodies, Anti-Idiotypic
  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Humanized
  • Antineoplastic Agents
  • Immunoglobulin G
  • Omalizumab
  • Trastuzumab