Chapter 11. Analysis of N- and O-Glycans of Glycoproteins by HPLC Technology

  1. Claus-Wilhelm von der Lieth3,
  2. Thomas Lütteke4 and
  3. Martin Frank3
  1. Anthony H. Merry1 and
  2. Sviatlana A. Astrautsova2

Published Online: 13 NOV 2009

DOI: 10.1002/9780470029619.ch11

Bioinformatics for Glycobiology and Glycomics: An Introduction

Bioinformatics for Glycobiology and Glycomics: An Introduction

How to Cite

Merry, A. H. and Astrautsova, S. A. (2009) Analysis of N- and O-Glycans of Glycoproteins by HPLC Technology, in Bioinformatics for Glycobiology and Glycomics: An Introduction (eds C.-W. von der Lieth, T. Lütteke and M. Frank), John Wiley & Sons, Ltd, Chichester, UK. doi: 10.1002/9780470029619.ch11

Editor Information

  1. 3

    Molecular Structure Analysis Core Facility, Deutsches Krebsforschungszentrum (German Cancer Research Center), Heidelberg, Germany

  2. 4

    Faculty of Veterinary Medicine, Institute of Biochemistry and Endocrinology, Justus-Liebig University Gießen, Gießen, Germany

Author Information

  1. 1

    Glycosciences Consultancy, Charlbury OX7 3HB, UK

  2. 2

    Department of Microbiology, Medical University of Grodno, Grodno, Belarus

Publication History

  1. Published Online: 13 NOV 2009
  2. Published Print: 11 DEC 2009

ISBN Information

Print ISBN: 9780470016671

Online ISBN: 9780470029619

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Keywords:

  • HPLC;
  • N glycans;
  • O glycans;
  • glycoproteins;
  • mass spectrometric;
  • exoglycosidase;
  • recombinant glycoproteins;
  • IgG

Summary

HPLC and capillary electrophoretic analysis of glycans provides a convenient means of analysis requiring no specialised technique or equipment. When used with fluorescent labelling it allows high sensitivity and quantitative analysis in the nanamolar range. Combined with techniques such as enzymatic sequencing or mass spectrometry it allows complete characterisation of glycans in biological materials. A guide to sample preparation and suitable chromatographic systems is given along with examples of glycans analysed in this way. A guide to data interpretation is also presented covering standardisation of the analysis and describing how complete structural analysis may be performed.