O-Linked Oligosaccharide Profiling by HPLC

  • Elizabeth F. Hounsell
  • Michael J. Davies
  • Kevin D. Smith
Part of the Springer Protocols Handbooks book series (SPH)


The several different core regions of O-linked chains (Chapter 105) can be further extended by Gal and GlcNAc containing backbones or by addition of blood group antigen-type glycosylation and the presence of sialic acid or sulfate. We sought a universal column that can be applied with high resolution to the various oligosaccharide alditols released from glycoproteins by β-elimination (see Note 1) and have pioneered (1,2) the use of porous graphitized carbon (PGC). This is an alternative to C18 reversed-phase HPLC and normal-phase amino-bonded columns, which can be used together in the presence and absence of high-salt buffers (3).


Sialic Acid Control Software Porous Graphitized Carbon Longe Retention Time Dynamic Mixer 
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  1. 1.
    Davies, M. J., Smith, K. D., Harbin, A-.M., and Hounsell, E. F. (1992) High performance liquid chromatography of oligosaccharide alditols and glycopeptides on graphitized carbon column. J. Chromatogr. 609, 125–131.PubMedCrossRefGoogle Scholar
  2. 2.
    Davies, M. J., Smith, K. D., Carruthers, R. A., Chai, W., Lawson, A. M., and Hounsell, E. F. (1993) The use of a porous graphitized carbon (PGC) column for the HPLC of oligosaccharides, alditols and glycopeptides with subsequent mass spectrometry analysis. J. Chromatogr. 646, 317–326.PubMedCrossRefGoogle Scholar
  3. 3.
    Davies, M. J. and Hounsell, E. F. (1996) Comparison of separation modes for high performance liquid chromatography of glycoprotein-and proteoglycan-derived oligosaccharides. J. Chromatogr. 720, 227–234.CrossRefGoogle Scholar
  4. 4.
    Hounsel, E. F., ed. (1993) Methods in Molecular Biology, vol 14: Glycoprotein Analysis in Biomedicine, Humana, Totowa, NJ.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Elizabeth F. Hounsell
    • 1
  • Michael J. Davies
    • 1
  • Kevin D. Smith
    • 2
  1. 1.Department of Biochemistry and Molecular BiologyUniversity College LondonUK
  2. 2.Department of Pharmaceutical SciencesUniversity of StrathclydeGlasgowScotland, UK

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