KCS

Protein glycosylation plays an important role in determining in vivo bioactivity, stability, solubility, and metabolic fate of biotherapeutics. Thus, detailed glycomic analyses are necessary complement to ensure quality, safety, and potency of therapeutic glycoproteins during production process. Current methods such as bottom-up and middle-down approaches employing LC-MS and LC-MS/MS for the glycomic characterization of biotherapeutics provide valuable information regarding glycan structure, isomers, and micro-heterogeneity. However, these methods significantly demand massive time and efforts from sample preparation to data interpretation. For these reasons, glycan or glycopeptide based approach is not suitable for real-time quality control in manufacturing processes. This study aims to develop novel analytical tool for real time determination of glycosylation site-occupancy with micro-heterogeneity using intact protein analysis (native mass spectrometry) without any additional sample preparation steps which can provide comprehensive glycosylation information.