Simultaneous glycosylation analysis of human serum glycoproteins by high-performance liquid chromatography/tandem mass spectrometry

J Chromatogr B Analyt Technol Biomed Life Sci. 2008 Jun 15;869(1-2):20-30. doi: 10.1016/j.jchromb.2008.05.006. Epub 2008 May 10.

Abstract

Changes in the glycosylation of some serum proteins are associated with certain diseases. In this study, we performed simultaneous site-specific glycosylation analysis of abundant serum glycoproteins by LC/Qq-TOF MS of human serum tryptic digest, the albumin of which was depleted. The glycopeptide peaks on the chromatogram were basically assigned by database searching with modified peak-list text files of MS/MS spectra and then based on mass differences of glycan units from characterized glycopeptides. Glycopeptide of IgG, haptoglobin and ceruloplasmin were confirmed by means of a comparison of their retention times and m/z values with those obtained by LC/MS of commercially available glycoproteins. Mass spectrometric carbohydrate heterogeneity in the assigned glycopeptides was analyzed by an additional LC/MS. We successfully demonstrated site-specific glycosylation of 23 sites in abundant serum glycoproteins.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid / methods*
  • Databases, Protein
  • Glycopeptides / analysis
  • Glycopeptides / isolation & purification
  • Glycoproteins / blood*
  • Glycoproteins / chemistry*
  • Glycoproteins / metabolism
  • Glycosylation
  • Humans
  • Protein Processing, Post-Translational
  • Tandem Mass Spectrometry / methods*

Substances

  • Glycopeptides
  • Glycoproteins