Identification of N-linked oligosaccharide labeled with 1-pyrenesulfonyl chloride by quadrupole time-of-flight tandem mass spectrometry after separation by micro- and nanoflow liquid chromatography
| Autor: | Takamasa Kurihara, Jun Zhe Min, Shinsuke Inagaki, Toshimasa Toyo'oka, Asuka Hirata |
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| Rok vydání: | 2009 |
| Předmět: |
Clinical Biochemistry
Analytical chemistry Oligosaccharides Pronase Tandem mass spectrometry Biochemistry Chloride Analytical Chemistry Acetylglucosamine Tandem Mass Spectrometry Drug Discovery medicine Nanotechnology Quadrupole time of flight Molecular Biology Chromatography High Pressure Liquid Pharmacology chemistry.chemical_classification Chromatography Pyrenes Elution Chemistry Galactose General Medicine Oligosaccharide Glycoprotein Mannose medicine.drug |
| Zdroj: | Biomedical chromatography : BMC. 23(9) |
| ISSN: | 1099-0801 |
| Popis: | The development of a qualitative determination method for the N-linked oligosaccharides in glycoproteins was attempted by the combination of micro- or nanoflow LC with Q-TOF-MS/MS. The asparaginyl-oligosaccharides in glycoproteins, liberated from the treatment of Pronase E, were separated, purified and labeled with 1-pyrenesulfonyl chloride (PSC). The resulting derivatives were separated by the microflow LC system utilizing a 0.5 mm diameter microcolumn or nanoflow LC system utilizing a 75 µm diameter chip column. The eluted N-linked oligosaccharide derivatives were then introduced into the Q-TOF-MS instrument and sensitively detected in the ESI+ mode. Several factors (i.e. fragmentor, skimmer, Vcap voltages and collision energy) affecting the sensitivity of Q-TOF-MS/MS detection were optimized in both the micro- and nanoflow LC systems. Various fragment ions based on the carbohydrate units appeared on the MS/MS spectra. Among the peaks, m/z 600.16 corresponding to PSC-labeled Asp-HexNAc is the most important one to identify the asparaginyl-oligosaccharide. The N-linked oligosaccharides in the ovalbumin were easily identified by the selected-ion chromatogram at m/z 600.16 by the MS/MS detection. Therefore, the identification of N-linked oligosaccharides in glycoproteins seems to be possible by the proposed micro- and nanoflow LC separations followed by the Q-TOF-MS/MS detection. Copyright © 2009 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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