Abstract
The detailed glycan structural analysis of glycoprotein is amenable to glycopeptide enrichment. Here, we develop a simple, effective and economical approach to enrich glycopeptides from proteolytically digested peptide mixtures by chromatographic column packed with graphite carbon and activated charcoal (G/A-column). Glycopeptide from ovalbumin was efficiently enriched by homemade G/A-column using liquid chromatography and the structure of glycopeptide was obtained by tandem mass spectrometry using Fourier transform ion cyclotron resonance mass spectrometry. The results in this study demonstrate that G/A-column can be used to enrich N-glycolpeptides and be benefit for online identification of glycopeptide using LC-MS.
Highlights
Various investigations concerning oligosaccharide analysis have been performed because of the importance ofHow to cite this paper: Xin, L., Hou, L., Zhou, Y., Wang, H.P. and Yu, H.M. (2015) Online Effective Identification of Glycopeptide Using Liquid Chromatography Combined with Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FTICRMS)
The result demonstrated that G/A-column could efficiently isolate the glycopeptides from the digestion mixtures of ovalbumin, and detailed glycan structural analysis was obtained by using tandem mass spectrometry
The solution of digestion was dried by Speedvac and dissolved in 1% formic acid to optimize the pH condition facilitating the glycopeptide binding to packing materials
Summary
How to cite this paper: Xin, L., Hou, L., Zhou, Y., Wang, H.P. and Yu, H.M. (2015) Online Effective Identification of Glycopeptide Using Liquid Chromatography Combined with Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FTICRMS). (2015) Online Effective Identification of Glycopeptide Using Liquid Chromatography Combined with Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FTICRMS). Liquid chromatography (LC) combined with MS can hit the mark that isolates and identifies glycopeptides online [5] [6]. Based on the similar properties, it is regarded that there are differences in adsorptions, signal intensity, S/N ratio, peptide binding and loading capacity between graphite carbon and activated charcoal [9] [10]. We develop a relatively stable, effective and economical approach to isolate and identify glycopeptides using liquid chromatography (LC) online combined with electrospray ionization Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS). The result demonstrated that G/A-column could efficiently isolate the glycopeptides from the digestion mixtures of ovalbumin, and detailed glycan structural analysis was obtained by using tandem mass spectrometry
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