Assessment of Chemical Modifications of Sites in the CDRs of Recombinant Antibodies: Susceptibility Vs. Functionality of Critical Quality Attributes

Overview

Journal MAbs

Specialty Allergy & Immunology

Date 2014 Jan 21

PMID 24441081

Citations 53

Authors

Markus Haberger

Katrin Bomans

Katharina Diepold

Michaela Hook

Jana Gassner

Tilman Schlothauer

Adrian Zwick

Christian Spick

Jochen Felix Kepert

Brigitte Hienz

Michael Wiedmann

Hermann Beck

Philipp Metzger

Michael Molhoj

Constanze Knoblich

Ulla Grauschopf

Dietmar Reusch

Patrick Bulau

Affiliations

Soon will be listed here.

Abstract

Modifications like asparagine deamidation, aspartate isomerization, methionine oxidation, and lysine glycation are typical degradations for recombinant antibodies. For the identification and functional evaluation of antibody critical quality attributes (CQAs) derived from chemical modifications in the complementary-determining regions (CDRs) and the conserved regions, an approach employing specific stress conditions, elevated temperatures, pH, oxidizing agents, and forced glycation with glucose incubation, was applied. The application of the specific stress conditions combined with ion exchange chromatography, proteolytic peptide mapping, quantitative liquid chromatography mass spectrometry, and functional evaluation by surface plasmon resonance analysis was adequate to identify and functionally assess chemical modification sites in the CDRs of a recombinant IgG1. LC-Met-4, LC-Asn-30/31, LC-Asn-92, HC-Met-100c, and HC Lys-33 were identified as potential CQAs. However, none of the assessed degradation products led to a complete loss of functionality if only one light or heavy chain of the native antibody was affected.

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