Chinese Journal of Chromatography ›› 2013, Vol. 31 ›› Issue (11): 1057-1063.DOI: 10.3724/SP.J.1123.2013.05043

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Analysis of N-glycosylated protein and N-glycans in human plasma by hydrophilic enrichment strategy

MA Cheng1,2, PAN Yiting1,2, ZHANG Qi3, WANG Jifeng2, QIAN Xiaohong1,2, YING Wantao2   

  1. 1. School of Life Science and Technology, Beijing Institute of Technology, Beijing 100081, China;
    2. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China;
    3. School of Biological Sciences, Nanyang Technological University, 637551, Singapore
  • Received:2013-05-27 Revised:2013-08-26 Online:2013-11-28 Published:2013-12-05

Abstract:

N-glycosylation of proteins is one of the most important post-translational modifications (PTM). Many diagnostic biomarkers and therapeutic targets are glycosylated proteins. However, it is still a big challenge to identify glycoproteins in human plasma, because of the high dynamic range and microheterogeneity of glycosylation. In this work, hydrophilic interaction liquid chromatography (HILIC) enrichment, high-pH reversed-phase prefractionation and high accurate mass spectrometry (MS) analysis were combined to profile the N-glycoproteins in human plasma. In total, 637 N-glycosites from 299 glycoproteins (protein groups) were identified. There were also 31 glycoforms recognized after HILIC enrichment and MS analysis. Among the results, 107 glycosylation sites (16.8%) are newly found. This study provided a simple and reliable strategy for exploring potential N-glycoprotein biomarkers from complicated biological systems.

Key words: N-glycan, N-linked glycoprotein, filter-aided sample preparation (FASP), hydrophilic interaction liquid chromatography (HILIC), reversed-phase liquid chromatography (RPLC)

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