Monosaccharide composition analysis of glycoproteins by isotope-tag method and capillary LC/ESI-MS

Yao Xue Xue Bao. 2005 Jan;40(1):43-8.

Abstract

Aim: To develop a rapid and sensitive method for monosaccharide composition analysis.

Methods: Glycoprotein was first hydrolyzed to monosaccharides, which were subsequently reacetylated (amino monosaccharides), tagged with 2-aminopyridine and then separated and monitored in selected ion mode by CapGCC-LC/MS. The use of tetradeuterium labeled-aminopyridyl-monosaccharides prepared by tagging monosacahrides with hexadeuterium labeled 2-aminopyridine as internal standards improved the linearity and reproducibility in quantification.

Results: This method was successfully applied to monosaccharide composition analysis of model glycoproteins, fetuin and erythropoietin down to 1 pmol monosaccharides.

Conclusion: This method has been shown to be highly sensitive and is applicable to monosaccharide composition analysis of glycoproteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Liquid / methods
  • Erythropoietin / chemistry
  • Glycoproteins / chemistry*
  • Hydrolysis
  • Isotope Labeling
  • Monosaccharides / analysis*
  • Sensitivity and Specificity
  • Spectrometry, Mass, Electrospray Ionization
  • alpha-Fetoproteins / chemistry

Substances

  • Glycoproteins
  • Monosaccharides
  • alpha-Fetoproteins
  • Erythropoietin