Magnetic Bead-based Hydrophilic Interaction Liquid Chromatography for Glycopeptide Enrichments

in Sciences Citation Index(SCI), 科學引文索引資料庫(SCI)
標題Magnetic Bead-based Hydrophilic Interaction Liquid Chromatography for Glycopeptide Enrichments
出版類型SCI(Sciences Citation Index)
AuthorsChia-Hao Yeh, 葉家豪, Shu-Hui Chen 陳淑慧, Ding-Tzai Li 李丁在, Hong-Ping Lin 林弘萍, Hung-Jen Huang 黃弘仁, Chang 張誌益, Wen-Ling Shih 施玟玲, Chi-Liang Chern 陳志良, Fong-Ku Shi 石峰鵠, & Liao Ming-Huei 徐睿良
出版日期2012 / 2
其他編號NSC 98-2113-M-020 -002 -MY2

Purification of glycopeptides prior to the analysis by mass spectrometry (MS) is demanded due to ion suppression effect during ionization caused by the co-presence of non-glycosylated peptides. Among various purification methods, hydrophilic interaction liquid chromatography (HILIC) has become a popular
method in recent years. In this work, we reported a novel magnetic bead-based zwitterionic HILIC (ZIC-HILIC) material which was fabricated by coating a zwitterionic polymer synthesized by spontaneous acid-catalyzed polymerization of 4-vinyl-pyridinium ethanesulfonate monomer on iron oxide magnetic nanoparticles. The resulting magnetic ZIC-HILIC nanoparticles were shown to provide high specificity
and high recovery yield (95–100%) for the enrichment of glycopeptides from a standard glycoprotein, fetuin, using a simple magnetic bar. In addition, we proposed a two-step HILIC enrichment strategy using magnetic ZIC-HILIC nanoparticles for a large scale analysis of glycoproteins in complex biological samples. Using this approach, we identified 85 N-glycosylation sites in 53 glycoproteins from urine samples. Two novel glycosylation sites on N513 of uromodulin and N470 of lysosomal alpha-glucosidase which have
not yet been reported were identified by two-step HILIC approach. Furthermore, all these identified sites were confirmed by studies conducted using PNGase F deglycosylation and 18O enzymatic labeling.

期刊名稱 Journal of Chromatography A
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