High-Performance Liquid Chromatography Mapping Method for the Structural Analysis of N-Glycans at the Molecular, Cellular, and Tissue Levels

Methods Mol Biol. 2022:2556:19-30. doi: 10.1007/978-1-0716-2635-1_2.

Abstract

Glycans on viruses are major molecules that mediate the communication of the viruses with host cells. Investigating the structure of glycans may be important in the characterization of viruses. In this regard, the high-performance liquid chromatography (HPLC) is a widely used analytical technique. We developed a multidimensional HPLC mapping method that identifies glycan structures. This is achieved by separating and purifying glycans using three types of HPLC columns and comparing the elution time data with known data. This method enables the easy profiling of glycans at the molecular, cellular, and tissue levels and can be used to study the glycans of virological samples such as virus and their hosts. It also enables the discrimination of isomers, which is difficult via mass spectrometry. In this paper, we describe in detail the principle and method of the structural analysis of N-type glycans using the proposed method.

Keywords: HPLC; N-glycan; glycoproteins; glycosylation profiles.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Isomerism
  • Mass Spectrometry
  • Polysaccharides*

Substances

  • Polysaccharides