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Weak affinity chromatography of small saccharides with immobilised wheat germ agglutinin and its application to monitoring of carbohydrate transferase activity

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Bioseparation

Abstract

In this work we have evaluated the potential to use wheat germ agglutinin(WGA) for weak affinity chromatography (WAC) of N-acetyl derivatives ofmono-, di-, tri- and tetrasaccharides. WGA was used as a ligand in a highperformance liquid affinity chromatography (HPLAC) system. Isocraticaffinity chromatography was conducted where similar N-acetyl saccharideswere separated according to their binding strength to WGA. Affinities areweak and lie typically in the mM range. For example, for3′sialyllactose, the dissociation constant (Kd) wasfound to be 2.4 mM at 8°C. It was interesting to note that theWGA–HPLC column can distinguish between the anomeric forms ofN-acetylglucosamine. Weak affinity chromatography with immobilised WGA wasused in an enzyme assay to detect the activity of GlcNAc-transferases.

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Authors and Affiliations

  1. Department of Natural Sciences, University of Kalmar, P.O.B. 905, S-391 29, Kalmar, Sweden

    Sten Ohlson, Maria Bergström & Lisa Leickt

  2. Neose Technologies, Inc., 102 Witmer Road, Horsham, PA, 19044, USA

    David Zopf

Authors
  1. Sten Ohlson
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  2. Maria Bergström
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  3. Lisa Leickt
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  4. David Zopf
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Ohlson, S., Bergström, M., Leickt, L. et al. Weak affinity chromatography of small saccharides with immobilised wheat germ agglutinin and its application to monitoring of carbohydrate transferase activity. Bioseparation 7, 101–105 (1998). https://doi.org/10.1023/A:1008073314855

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  • DOI: https://doi.org/10.1023/A:1008073314855

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