Change of charge variant composition of trastuzumab upon stressing at physiological conditions

Baubek Spanov, Oladapo Olaleye, Nico Lingg, Arthur E H Bentlage, Natalia Govorukhina, Jos Hermans, Nico van de Merbel, Gestur Vidarsson, Alois Jungbauer, Rainer Bischoff*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Cation-exchange chromatography is a widely used approach to study charge heterogeneity of monoclonal antibodies. Heterogeneity may arise both in vitro and in vivo because of the susceptibility of monoclonal antibodies to undergo chemical modifications. Modifications may adversely affect the potency of the drug, induce immunogenicity or affect pharmacokinetics. In this study, we evaluated the application of optimized pH gradient systems for the separation of charge variants of trastuzumab after forced degradation study. pH gradient-based elution resulted in high-resolution separation of some 20 charge variants after 3 weeks at 37°C under physiological conditions. The charge variants were further characterized by LC-MS-based peptide mapping. There was no significant difference in the binding properties to HER2 or a range of Fcγ receptors between non-stressed and stressed trastuzumab.

Original languageEnglish
Article number462506
Number of pages9
JournalJournal of Chromatography A
Volume1655
Early online date28-Aug-2021
DOIs
Publication statusPublished - 11-Oct-2021

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