Oxidation in the complementarity-determining regions differentially influences the properties of therapeutic antibodies.
Therapeutic antibodies can undergo a variety of chemical modification reactions in vitro. Depending on the site of modification, either antigen binding or Fc-mediated functions can be affected. Oxidation of tryptophan residues is one of the post-translational modifications leading to altered antibody functionality. In this study, we examined the structural and functional properties of a therapeutic antibody construct and 2 affinity matured variants thereof. Two of the 3 antibodies carry an oxidation-prone tryptophan residue in the complementarity-determining region of the VLdomain. We demonstrate the differences in the stability and bioactivity of the 3 antibodies, and reveal differential degradation pathways for the antibodies susceptible to oxidation.
a Roche Pharma Research and Early Development (pRED), Large Molecule Research, Roche Innovation Center Munich , Germany.|b Center for Integrated Protein Science Munich , Department Chemie, Technische Universität München , Garching , Germany.
Tetyana
T
Dashivets
c Early-Stage Pharmaceutical Development & GLP Supplies, F. Hoffmann-La Roche Ltd Pharmaceutical Development & Supplies PTD Biologics Europe , Basel , Switzerland.
Jan
J
Stracke
a Roche Pharma Research and Early Development (pRED), Large Molecule Research, Roche Innovation Center Munich , Germany.
Stefan
S
Dengl
a Roche Pharma Research and Early Development (pRED), Large Molecule Research, Roche Innovation Center Munich , Germany.
Alexander
A
Knaupp
d Roche Pharma Biotech Penzberg , Germany.
Jan
J
Pollmann
b Center for Integrated Protein Science Munich , Department Chemie, Technische Universität München , Garching , Germany.
Johannes
J
Buchner
a Roche Pharma Research and Early Development (pRED), Large Molecule Research, Roche Innovation Center Munich , Germany.
Tilman
T
Schlothauer
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000915:Antibody Affinity / Q000276:immunology; D022801:Complementarity Determining Regions / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D010084:Oxidation-Reduction; D011499:Protein Processing, Post-Translational / Q000276:immunology*; D055550:Protein Stability
D000911:Antibodies, Monoclonal; D022801:Complementarity Determining Regions
Deamidation; monoclonal antibody; oxidation; posttranslational modifications; stability; succinimide; target binding