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Updated: Jan 27, 2026

Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry
Published on: July 20, 2016
Mechanistic investigation and further optimization of the aqueous Glaser-Hay bioconjugation
Christopher R Travis1, Lauren E Mazur, Emily M Peairs
1Department of Chemistry, College of William & Mary, PO Box 8795, Williamsburg, VA, USA 23185. dyoung01@wm.edu.
Abstract:
The Glaser-Hay bioconjugation has recently emerged as an efficient and attractive method to generate stable, useful bioconjugates with numerous applications, specifically in the field of therapeutics. Herein, we investigate the mechanism of the aqueous Glaser-Hay coupling to better understand optimization strategies. In doing so, it was identified that catalase is able to minimize protein oxidation and improve coupling efficiency, suggesting that hydrogen peroxide is produced during the aqueous Glaser-Hay bioconjugation. Further, several new ligands were investigated to minimize protein oxidation and maximize coupling efficiency. Finally, two novel strategies to streamline the Glaser-Hay bioconjugation and eliminate the need for secondary purification have been developed.
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