Simultaneous microanalysis of N-linked oligosaccharides in a glycoprotein using microbore graphitized carbon column liquid chromatography-mass spectrometry.
We previously reported that graphitized carbon column liquid chromatography-mass spectrometry (GCC-LC-MS) is very useful for the structural analysis of carbohydrates in a glycoprotein. In this study, GCC-LC-MS was adapted for the simultaneous microanalysis of oligosaccharides. A variety of oligosaccharide alditols prepared from fetuin, ribonuclease B, and recombinant human erythropoietin were used as model oligosaccharides. The use of microbore GCC-LC-MS was found to be successful for rapid, sensitive, and simultaneous analysis of high-mannose-type, desialylated fucosyl complex-type, sialylated complex-type, and sialylated fucosyl complex-type oligosaccharide alditols. Furthermore, we demonstrate that this method is applicable to the analysis of carbohydrate heterogeneity in a glycoprotein that possesses diverse oligosaccharides. Microbore GCC-LC-MS was able to characterize high-mannose-type, hybrid-type, and complex-type oligosaccharides in tissue plasminogen activator produced from human melanoma cells in a single analysis.
Itoh
Satsuki
S
Division of Biological Chemistry and Biologicals, National Institute of Health Science, Tokyo, Japan.
Kawasaki
Nana
N
Ohta
Miyako
M
Hyuga
Masashi
M
Hyuga
Sumiko
S
Hayakawa
Takao
T
D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry*; D006108:Graphite; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis*
D006023:Glycoproteins; D009844:Oligosaccharides; D006108:Graphite