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Simultaneous microanalysis of N-linked oligosaccharides in a glycoprotein using microbore graphitized carbon column liquid chromatography-mass spectrometry.

Satsuki Itoh·Nana Kawasaki·Miyako Ohta·Masashi Hyuga·Sumiko Hyuga·Takao Hayakawa
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Abstract

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.

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Carbohydrate SequenceChromatography, High Pressure LiquidGlycoproteinsGraphiteMass SpectrometryMolecular Sequence DataOligosaccharidesJournal Article

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Satsuki Itoh, Nana Kawasaki, Miyako Ohta, Masashi Hyuga, Sumiko Hyuga, Takao Hayakawa. 2002-08-30. Simultaneous microanalysis of N-linked oligosaccharides in a glycoprotein using microbore graphitized carbon column liquid chromatography-mass spectrometry.. https://doi.org/10.1016/s0021-9673(02)00951-2

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