Identifying Specific and Differentially Linked Glycosyl Residues in Mammalian Glycans by Targeted LC-MS Analysis

Anal Sci. 2018 Sep 10;34(9):1049-1054. doi: 10.2116/analsci.18SCP01. Epub 2018 Aug 24.

Abstract

Glycans, which are widespread in nature, consist of a large number of monosaccharides linked via glycosidic bonds. Due to the complex nature of glycan structures on glycoproteins, assessing the configuration and positions of the glycosidic linkages of a glycan is a subject of considerable interest. In this study, a method for accomplishing this using partially O-methylated alditols (PMAs) from glycans combined with LC-MS analysis is reported. N-Glycans were first per-methylated with methyl iodide, and the levels of methylation were further confirmed by MALDI-TOF. PMAs were then produced via complete hydrolysis and reduction. PMAs derived from Fetuin N-glycan and Lewisa antigen carbohydrates were successfully detected by LC-MS analysis. This analysis can be performed without the need for an additional derivatization step for GC analysis, and should be suitable for use in conjunction with a LC-MS-based analysis platform.

Keywords: LC-MS; Linkage analysis; partially methylated alditol.

MeSH terms

  • Acetates / chemistry
  • Animals
  • Chromatography, Liquid / methods*
  • Glycosylation
  • Methylation
  • Polysaccharides / chemistry*
  • Tandem Mass Spectrometry / methods*

Substances

  • Acetates
  • Polysaccharides