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The Protein Protocols Handbook pp 917–931Cite as

Affinity Chromatography of Oligosaccharides and Glycopeptides with Immobilized Lectins

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Abstract

Sugar moieties on the cell surface play one of the most important roles in cellular recognition. To elucidate the molecular mechanism of these cellular phenomena, assessment of the structure of sugar chains is indispensable. However, it is difficult to elucidate the structures of cell surface oligosaccharides owing to two technical problems. The first is the difficulty in fractionating various oligosaccharides heterogeneous in the number, type, and substitution patterns of outer sugar branches. The second problem is that very limited amounts of material can be available, which makes it difficult to perform detailed structural studies. Lectins are proteins with sugar binding activity. Each lectin binds specifically to a certain sugar sequence in oligosaccharides and glycopeptides. To overcome these problems, lectins can serve as very useful tools. Recently, many attempts have been made to fractionate oligosaccharides and glycopeptides on immobilized lectin columns. The use of a series of immobilized lectin columns, whose sugar binding specificities have been precisely elucidated, enables us to fractionate a very small amount of radioactive oligosaccharides or glycopeptides (∼10 ng depending on the specific activity) into structurally distinct groups. In this chapter, we summarize the serial lectin-Sepharose affinity chromatographic technique for rapid, sensitive, and specific fractionation and analysis of asparagine-linked oligosaccharides of glycoproteins.

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Author information

Authors and Affiliations

  1. Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Japan

    Kazuo Yamamoto

  2. Hoshi Pharmaceutical College, Tokyo, Japan

    Tsutomu Tsuji

  3. Yakult Central Institute for Microbiology Research, Tokyo, Japan

    Toshiaki Osawa

Authors
  1. Kazuo Yamamoto
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  2. Tsutomu Tsuji
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  3. Toshiaki Osawa
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Editor information

Editors and Affiliations

  1. University of Hertfordshire, Hatfield, UK

    John M. Walker

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© 2002 Humana Press Inc., Totowa, NJ

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Yamamoto, K., Tsuji, T., Osawa, T. (2002). Affinity Chromatography of Oligosaccharides and Glycopeptides with Immobilized Lectins. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-169-8:917

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  • DOI: https://doi.org/10.1385/1-59259-169-8:917

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-940-7

  • Online ISBN: 978-1-59259-169-5

  • eBook Packages: Springer Book Archive

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