Glycobiology Protocols pp 125-143

Part of the Methods in Molecular Biology book series (MIMB, volume 347)

Detailed Structural Analysis of N-Glycans Released From Glycoproteins in SDS-PAGE Gel Bands Using HPLC Combined With Exoglycosidase Array Digestions

  • Louise Royle
  • Catherine M. Radcliffe
  • Raymond A. Dwek
  • Pauline M. Rudd

Abstract

In contrast to the linear sequences of protein and DNA, oligosaccharides are branched structures. In addition, almost all glycoproteins consist of a heterogeneous collection of differently glycosylated variants. Glycan analysis therefore requires high-resolution separation techniques that can provide detailed structural analysis, including both monosaccharide sequence and linkage information. This chapter describes how a combination of high-performance liquid chromatography (HPLC) and exoglycosidase enzyme array digestions can deliver quantitative glycan analysis of sugars released from glycoproteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel bands by matching HPLC elution positions with a database of standard glycans.

Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Louise Royle
    • 1
  • Catherine M. Radcliffe
    • 1
  • Raymond A. Dwek
    • 1
  • Pauline M. Rudd
    • 1
  1. 1.Department of Biochemistry, Glycobiology InstituteUniversity of OxfordOxfordEngland

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