HPLC Strategies for Profiling and Sequencing Oligosaccharides

  • Geoffrey R. Guile
  • Pauline M. Rudd
  • David R. Wing
  • Raymond A. Dwek
Part of the BioMethods book series (BIOMETHODS)


The sugars released from a pure glycoprotein often consist of a heterogeneous population containing both neutral and charged oligosaccharides. For example, the single N-glycosylation site in human erythrocyte CD59 is associated with more than 100 neutral and sialylated complex glycans, each representing a different glycoform (1). The existence of such extensive heterogeneity in biologically important glycoproteins requires refined approaches to the analysis of oligosaccharides. The adaptable technology which is described here represents a significant advance towards faster, more automated and more detailed strategies for the rapid profiling and analysis of sugars. Such technologies may be required for major studies, such as the human genome project, which defines DNA in normal and diseased states, and the proteome project, which sets out to analyse the total amount of protein in a living cell. It is worthy of note that genetic diseases are not caused by the genes themselves, but by the products for which the genes code and their post-translational modifications, which include glycosylation. In this chapter two strategies for rapid oligosaccharide analysis are described: Oligosaccharide profiling and detailed structural analysis.


Sialic Acid Normal Phase Glucose Unit Jack Bean Normal Phase HPLC 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Rudd PM, Morgan BP, Wormald MR, Harvey DJ, Van der Berg CW, Davis SJ et al. (1997) J. BioLChem (in press).Google Scholar
  2. 2.
    Bigge JC, Patel TP, Bruce JA, Goulding PN, Charles SM and Parekh RB (1995) Anal. Biochem. 230: 229.PubMedCrossRefGoogle Scholar
  3. 3.
    Ashford D, Dwek RA, Welply J K, Amatayakul S, Homans S, Lis H et al. (1987) Eur. J. Biochem. 166: 311.PubMedCrossRefGoogle Scholar
  4. 4.
    Wing DR., Rademacher TW, Field MC, Dwek RA, Schmitz B, Thor G et al. (1992) Glycoconjugate J. 9: 293.CrossRefGoogle Scholar
  5. 5.
    Guile GR, Wong SYC, and Dwek RA (1994) Anal. Biochem. 222: 231.PubMedCrossRefGoogle Scholar
  6. 6.
    Green ED, Adelt G, Baenziger JU, Wilson, Sand Van Halbeek H (1988) J. Biol Chem. 263: 18253.PubMedGoogle Scholar
  7. 7.
    Jacob GS and Scudder P (1994) Methods Enzymol. 230: 280.PubMedCrossRefGoogle Scholar
  8. 8.
    Parekh RB, Dwek RA, Sutton BJ, Fernandes DL, Leung A, Stanworth D et al. (1985) Nature 316: 452.PubMedCrossRefGoogle Scholar
  9. 9.
    Iourin O, Mattu TS, Mian N, Keir G, Winchester B, Dwek RA et al. (1996) Glycoconjugate J. 13: 1031.CrossRefGoogle Scholar
  10. 10.
    Guile GR, Rudd PM, Wing DR, Prime SB and Dwek RA (1996) Anal. Biochem. 240: 210.PubMedCrossRefGoogle Scholar
  11. 11.
    Harvey DJ and Rudd PM, Bateman RH, Bordoli RS, Howes K, Hoyes JB et al. (1994) Organic Mass Spectrom. 29: 753.CrossRefGoogle Scholar
  12. 12.
    Rudd PM (1995) PhD Thesis, The Open University, Milton Keynes, UK.Google Scholar

Copyright information

© Birkhäuser Verlag Basel 1997

Authors and Affiliations

  • Geoffrey R. Guile
  • Pauline M. Rudd
  • David R. Wing
  • Raymond A. Dwek

There are no affiliations available

Personalised recommendations