Assessment of monoclonal antibody glycosylation: a comparative study using HRMS, NMR, and HILIC-FLD

In this study, two products were compared using a panel of analytical methods. Two high-resolution mass spectrometry (HRMS) workflows were used to analyze N-glycans, while nuclear magnetic resonance (NMR) was used to generate monosaccharide fingerprints. These state-of-the-art techniques were compared to conventional analysis using hydrophilic interaction chromatography (HILIC) coupled with fluorescence detection (FLD). The advantages and disadvantages of each method are discussed along with a comparison of the identified glycan distributions. The results demonstrated agreement across all methods for major glycoforms, demonstrating how confidence in glycan characterization is increased by combining orthogonal analytical methodologies. The full panel of methods used represents a diverse toolbox that can be selected from based on the needs for a specific product or analysis.PMID:38580890 | DOI:10.1007/s00216-024-05261-5

Source: Analytical and Bioanalytical Chemistry - April 5, 2024 Category: Chemistry Authors: Joshua Shipman Michael Karfunkle Hongbin Zhu You Zhuo Kang Chen Milani Patabandige Di Wu Mercy Oyugi Richard Kerr Kui Yang Sarah Rogstad Source Type: research