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N-Linked Oligosaccharide Profiling by HPAEC-PAD

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

Abstract

High-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) is a very powerful tool for the profiling of N-linked oligosaccharides (1,2), only being limited by the concentrated sodium hydroxide and sodium acetate required to achieve the separation and sensitive detection. Oligosaccharides are separated on the basis of charge (i.e., the number of sialic acid residues) and the linkage isomers present. Neutral oligosaccharide alditols are most weakly retained, retention increasing with increasing sialylation, and NeuGc- and sulfate-bearing oligosaccharides being most strongly retained. Fucosylation results in a shorter retention. If the oligosaccharides can be effectively desalted after HPAEC-PAD, this remains the method of choice for oligosaccharide purification, and is a very powerful analytical tool.

Keywords

Sodium Acetate Sialic Acid Residue Pulse Amperometric Detection Powerful Analytical Tool Dynamic Mixer 
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.
    Townsend, R. R. (1995) Analysis of glycoconjugates using high pH anion-exchange chromatography. J. Chromatogr. Library 58, 181–209.CrossRefGoogle Scholar
  2. 2.
    Smith, K. D., Davies, M. J., and Hounsell, E. F. (1994) Structural profiling of oligosaccharides of glycoproteins, in Methods in Molecular Biology, vol. 32: Basic Protein and Peptide Protocols (Walker, J. M., ed.), Humana, Totowa, NJ, pp. 143–155.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Elizabeth F. Hounsell
    • 1
  • Michael J. Davies
  • Kevin D. Smith
  1. 1.School of Biological and Chemical SciencesBirkbeck University of LondonUK

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