Magnetic bead-based hydrophilic interaction liquid chromatography for glycopeptide enrichments

Chia Hao Yeh, Shu-Hui Chen, Ding Tzai Li, Hong-Ping Lin, Hung Jen Huang, Chi I. Chang, Wen Ling Shih, Chi Liang Chern, Fong Ku Shi, Jue Liang Hsu

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

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 N 513 of uromodulin and N 470 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.

Original languageEnglish
Pages (from-to)70-78
Number of pages9
JournalJournal of Chromatography A
Volume1224
DOIs
Publication statusPublished - 2012 Feb 10

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Fingerprint Dive into the research topics of 'Magnetic bead-based hydrophilic interaction liquid chromatography for glycopeptide enrichments'. Together they form a unique fingerprint.

  • Cite this

    Yeh, C. H., Chen, S-H., Li, D. T., Lin, H-P., Huang, H. J., Chang, C. I., Shih, W. L., Chern, C. L., Shi, F. K., & Hsu, J. L. (2012). Magnetic bead-based hydrophilic interaction liquid chromatography for glycopeptide enrichments. Journal of Chromatography A, 1224, 70-78. https://doi.org/10.1016/j.chroma.2011.12.057