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


CONCLUSIONS
This method has been shown to be highly sensitive and is applicable to monosaccharide composition analysis of glycoproteins.
Yuan
Jin
J
Gene Engineering Department, Chengdu Institute of Biological Products, Chengdu 610063, China. yuanjinv@hotmail.com
Kawasaki
Nana
N
Hashii
Noritaka
N
Itoh
Satsuki
S
Kawanishi
Toru
T
Hayakawa
Takao
T
D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D004921:Erythropoietin / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry*; D006868:Hydrolysis; D007553:Isotope Labeling; D009005:Monosaccharides / Q000032:analysis*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D000509:alpha-Fetoproteins / Q000737:chemistry
D006023:Glycoproteins; D009005:Monosaccharides; D000509:alpha-Fetoproteins; D004921:Erythropoietin