Monosaccharide composition analysis of glycoproteins by isotope-tag method and capillary LC/ESI-MS.
- Author:
Jin YUAN
1
;
Nana KAWASAKI
;
Noritaka HASHII
;
Satsuki ITOH
;
Toru KAWANISHI
;
Takao HAYAKAWA
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Chromatography, Liquid; methods; Erythropoietin; chemistry; Glycoproteins; chemistry; Hydrolysis; Isotope Labeling; Monosaccharides; analysis; Sensitivity and Specificity; Spectrometry, Mass, Electrospray Ionization; alpha-Fetoproteins; chemistry
- From: Acta Pharmaceutica Sinica 2005;40(1):43-48
- CountryChina
- Language:English
-
Abstract:
AIMTo develop a rapid and sensitive method for monosaccharide composition analysis.
METHODSGlycoprotein 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.
RESULTSThis method was successfully applied to monosaccharide composition analysis of model glycoproteins, fetuin and erythropoietin down to 1 pmol monosaccharides.
CONCLUSIONThis method has been shown to be highly sensitive and is applicable to monosaccharide composition analysis of glycoproteins.
