Recent Advances in the Analysis of Protein Glycosylation by Hydrophilic Interaction Liquid Chromatography-Mass Spectrometry

The recent advances in bioanalytical technology have accelerated the analysis of complex protein glycosylation, which is beneficial to understanding glycosylation in drug discovery and disease diagnosis. Due to its biological uniqueness in the course of disease occurrence and development, disease-specific glycosylation requires quantitative characterization. The article comprehensively reviewed the latest development of glycosylation and primary procedures including digestion of glycoproteins, separation and enrichment of glycopeptides, isolation of N-glycopeptide, enrichment of O-glycans and O-glycopeptides, and mass spectrometry sequencing of glycopeptides. It focuses on the technological innovation of hydrophilic interaction media for glycopeptide enrichment and separation, and the improvement of the identification of intact glycopeptides using various mass spectrometry fragmentation techniques. This review also discusses the challenges of glycosylation analysis and provides some advanced media for the development of hydrophilic interaction liquid chromatography using carbohydrates and glycopeptides to enrich glycosylation, and use O-proteases to analyze O-glycosylation