Journal
JOURNAL OF SEPARATION SCIENCE
Volume 34, Issue 19, Pages 2745-2750Publisher
WILEY-BLACKWELL
DOI: 10.1002/jssc.201100427
Keywords
C18 fractionation; Glycopeptide; Hydrophilic interaction chromatography; Mass spectrometry; TiO(2) enrichment
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Funding
- National Science Foundation of China [30801513, 20975100]
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Comprehensive glycoprotein characterization based on mass spectrometry (MS) is challenging because of low concentration of glycopeptides and suppression effect of abundant non-glycosylated peptides in MS. Therefore, it is vital to enrich glycopeptides before MS analysis. A new method was developed to selectively enrich glycopeptides from complex sample by coupling C18 fractionation with titanium dioxide (TiO(2)) enrichment. The new method allows to selectively enrich N-linked glycopeptides with various glycan forms and different sequence lengths. Compared with single TiO(2) method, the established method demonstrated higher glycopeptide selectivity and higher glycosylation heterogeneity coverage. Further application of this method to mixture of non-glycosylated protein and glycoprotein digests at different levels reveals the feasibility of enrichment of tryptic glycopeptides from simple proteomics samples.
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