Journal
ANALYTICA CHIMICA ACTA
Volume 1058, Issue -, Pages 107-116Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2019.01.044
Keywords
Intact tryptic N-glycopeptide; Enrichment; Size exclusion chromatography; Hydrophilic interaction liquid chromatography; Acrylamide-agarose composite gel; Formalin-fixed and paraffin-embedded
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Funding
- National Key Research and Development Program [2016YFA0501303, 2018YFC0910300]
- NSF of China [21335002, 31670835]
- Programs Foundation of Ministry of Education of China [20130071110034]
- Key Laboratory of Glycoconjugates Research Ministry of Public Health
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Increasing researches proved that abnormal glycosylation is strongly correlated with many diseases. Specially, site-specific glycosylation and its associated heterogeneity are closely related to the function and activity of the glycoprotein. However, intact N-glycopeptide analysis still faces great challenges because the presence of highly abundant non-glycosylated peptides would suppress the ionization of lowly abundant glycopeptides. In the present study, we developed a practical intact tryptic N-glycopeptide enrichment method using acrylamide-agarose composite gel that combined the size exclusion chromatography and hydrophilic (named SELIC) effects, aimed to remove the detergent rapidly and effectively, as well as enrich intact N-glycopeptides while extracting peptides. This is a useful tool to facilitate the intact N-glycopeptides analysis of complex protein mixtures, particularly for samples that extracted from formalin-fixed and paraffin-embedded (FFPE) tissues by SDS. Using this method, we successfully identified 700 site-specific intact tryptic N-glycopeptides corresponding to 261 glycosylation sites on 191 glycoproteins from FFPE thymoma tissues. (C) 2019 Elsevier B.V. All rights reserved.
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