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Simultaneous Fluorescent Labelling and Biotinylation of Oligosaccharides: A Versatile Approach to the Analysis of Oligosaccharide Structure and Function

  • Derek K. Toomre
  • Ajit Varki
Part of the BioMethods book series (BIOMETHODS)

Abstract

Oligosaccharides participate in a variety of important biological functions, including interactions with specific receptors (1–4). A prerequisite to the discovery and exploration of such interactions is the comprehensive structural analysis of homogeneous oligosaccharide species. The purification of oligosaccharides from complex mixtures requires diverse techniques, as well as sensitive and specific detection. One way to achieve sensitive detection is to attach a chromophore to the reducing sugar by reductive amination (5, 6) (see Chapters 7,11,15,18 in this volume for other examples of such techniques). Fluorescent chromophores (in contrast to light-absorbing chromophores) can often permit detection in the low picomole range for high-pressure liquid chromatography (HPLC) and polyacrylamide gel electrophoresis (PAGE) techniques, and in the femtomole range in capillary electrophoresis. Furthermore, hydrophobic or anionic properties of the chromophoric groups can be exploited to improve oligosaccharide fractionation by either electrophoresis or HPLC (5–8). Often several multidimensional chromatographic techniques, based on different physical properties, are required to adequately resolve complex mixtures. Several examples have been described of “two-dimensional mapping” with fluorescent pyridylamino (PA)-coupled oligosaccharides (5, 9). More recently, 2-aminobenzamide has been employed as an alternate fluorescent tag which gives both nonselective and highly efficient coupling to oligosaccharides (10).

Keywords

Ammonium Formate Reductive Amination Pyridine Acetate Fast Atom Bombard Troubleshooting Guide 
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.

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Copyright information

© Birkhäuser Verlag Basel 1997

Authors and Affiliations

  • Derek K. Toomre
  • Ajit Varki

There are no affiliations available

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