Quantification of Glycan in Glycoproteins via AUCAgent-Enhanced Analytical Ultracentrifugation

TL;DR

This paper introduces a new method using analytical ultracentrifugation to accurately measure glycans in glycoproteins, important for vaccine development.

Contribution

A novel glycan quantification method using AUC with UV absorption and interference data is introduced.

Findings

The new method enables precise glycan mass fraction determination in glycoproteins.

The approach retains AUC's advantages with high reproducibility and native state analysis.

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.

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