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Updated: Feb 2, 2026

A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples
Published on: October 1, 2009
Multi-arm PEG-maleimide conjugation intermediate characterization and hydrolysis study by a selective HPLC method
Jenny Wang1, Samuel Yang1, Kelly Zhang1
1Research and Early Development, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, United States.
Abstract:
Maleimide functionalized polyethylene glycol (PEG) with varying degrees of branched chains is gaining increasing interest in drug conjugation. Characterization of these molecules in solution is highly challenging due to the multiplicity of the hydrolysis products and the subtle differences between them, especially for high molecular weight PEG-maleimide. A highly selective, sensitive and stability indicating HPLC method employing charged surface hybrid (CSH) stationary phase was developed for 8-arm PEG-maleimide characterization. The method has been validated per ICH Q2 guideline for specificity, sensitivity, linearity, accuracy and precision for quality and process control. Hydrolysis of 8-arm PEG-maleimide in different buffer media, pH and temperature was studied. The kinetics revealed the hydrolysis is a first-order reaction. Hydrolysis rate constant and half-life were obtained to guide protein conjugation process development.
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