期刊
TALANTA
卷 146, 期 -, 页码 225-230出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.talanta.2015.08.037
关键词
Amide functionalized monolith; Hydrophilic interaction liquid chromatography; Glycopeptide enrichment; Glycoproteomics
资金
- National Basic Research Program of China [2012CB910601, 2013CB911201]
- National Natural Science Foundation of China [21190043, 21175131, 21205115]
- Creative Research Group Project by National Natural Science Foundation of China [21321064]
In this study, a novel kind of amide functionalized hydrophilic monolith was synthesized by the in situ photo-polymerization of N-vinyl-2-pyrrolidinone (NVP), acrylamide (AM), and N, N'-methylenebisacrylamide (MBA) in a UV transparent capillary, and successfully applied for hydrophilic interaction chromatography (HILIC) based enrichment of N-linked glycopeptides. With 2 mu g of the tryptic digests of IgG as the sample, after enrichment, 18 glycopeptides could be identified by MALDI-TOF/TOF MS analysis. Furthermore, with the mixture of BSA and IgG digests ( 10,000:1, m/m) as the sample, 6 N-linked glycopeptides were unambiguously identified after enrichment, indicating the high selectivity and good specificity of such material. Moreover, such a monolithic capillary column was also applied for the N-glycosylation sites profiling of 614 protein digests from HeLa cells and I mu L human serum. In total, 530 and 262 unique N-glycosylated peptides were identified, respectively, corresponding to 282 and 124 N-glycoproteins, demonstrating its great potential for the large scale glycoproteomics analysis. (C) 2015 Elsevier B.V. All rights reserved.
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