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Chemical Methods of Analysis of Glycoproteins

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

Abstract

The first analysis of glycoconjugates that often needs to be carried out is to see if they indeed contain sugar. For glycoproteins in gels or oligosaccharides in solution, this can be readily achieved by periodate oxidation at two concentrations, the first to detect sialic acids, and the second, any monosaccharide that has two free vicinal hydroxyl groups (1). Periodate cleaves between the hydroxyl groups to yield reactive aldehydes, which can be detected by reduction with NaB3H4 or coupled to high sensitivity probes available in commercial kits, e.g., from Boeringher Mannheim (Mannheim, Germany) or Oxford Glycosystems (Abingdon, UK). In solution, a quick spot assay can be carried out for the presence of any monosaccharide or oligosaccharide having a C-2 hydroxyl group by visualization with charring by phenol/sulfuric acid reagent (2). These methods are relatively specific for mono/oligosaccharides (3).

Keywords

Sialic Acid Sodium Borohydride Acid Reagent Periodate Oxidation Microtiter Plate Reader 
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.

References

  1. 1.
    Haselbeck, A. and Hösel, W. (1993) Immunological detection of glycoproteins on blots based on labeling with digoxigenin, in Methods in Molecular Biology, vol. 14: Glycoprotein Analysis in Biomedicine (Hounsell, E. F., ed.), Humana, Totowa, NJ, pp. 161–173.CrossRefGoogle Scholar
  2. 2.
    Smith, K., Harbin, A. M., Carruthers, R. A., Lawson, A. M., and Hounsell, E. F. (1990) Enzyme degradation, high performance liquid chromatography and liquid secondary ion mass spectrometry in the analysis of glycoproteins. Biomed. Chromatogr. 4, 261–266.PubMedCrossRefGoogle Scholar
  3. 3.
    Hounsell, E. F. (1993) A general strategy for glycoprotein oligosaccharide analysis, in Methods in Molecular Biology, vol. 14: Glycoprotein Analysis in Biomedicine (Hounsell, E. F., ed.), Humana, Totowa, NJ, pp. 1–15.CrossRefGoogle 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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