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Updated: Feb 28, 2026
Immunoglobulin G N-Glycan Analysis by Ultra-Performance Liquid Chromatography
Published on: January 18, 2020
Quantification of Glycan in Glycoproteins via AUCAgent-Enhanced Analytical Ultracentrifugation
Xiaojuan Yu1, Zhaoxing Wang2,3, Chengshi Zeng2
1State Key Laboratory of Drug Regulatory Science, NHC Key Laboratory of Research on Quality and Standardization of Biotech Products, NMPA Key Laboratory for Quality Research and Evaluation of Biological Products, National Institutes for Food and Drug Control, Beijing 102629, China.
Abstract:
Background: As essential biomolecules composed of proteins and carbohydrate moieties, glycoproteins play pivotal roles in numerous biological processes. The glycosylation level plays a crucial role in determining the functionality of glycoproteins. Therefore, the precise quantification of glycan components in proteins holds significant importance for research on and development of polysaccharide-protein-conjugated vaccines. Methods: In this study, a novel glycan quantification approach was developed, leveraging analytical ultracentrifugation (AUC) technology that synergistically utilizes ultraviolet wavelength absorption and interference data to directly determine glycan mass fractions in glycoproteins. Results: This methodology expands the analytical framework for glycoproteins while retaining the intrinsic advantages of AUC, enabling analysis in native states with high reproducibility as indicated by low standard deviation across replicates. Conclusions: The approach was implemented in our proprietary AUC data analysis software called AUCAgent (v1.8.8), providing a new method for glycoprotein quantification and polysaccharide ratio determination in polysaccharide-protein-conjugate vaccines.
