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    Combined strong anion-exchange HPLC and PAGE approach for the purification of heparan sulphate oligosaccharides.

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    Authors
    Vivès, Romain R
    Goodger, Sarah J
    Pye, David A
    Affiliation
    CRC Drug Development Group, and University of Manchester Department of Medical Oncology, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, UK.
    Issue Date
    2001-02-15
    
    Metadata
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    Abstract
    Heparan sulphates are highly sulphated linear polysaccharides involved in many cellular functions. Their biological properties stem from their ability to interact with a wide range of proteins. An increasing number of studies, using heparan sulphate-derived oligosaccharides, suggest that specific structural features within the polysaccharide are responsible for ligand recognition and regulation. In the present study, we show that strong anion-exchange HPLC alone, a commonly used technique for purification of heparan sulphate-derived oligosaccharides, may not permit the isolation of highly pure heparan sulphate oligosaccharide species. This was determined by PAGE analysis of hexa-, octa- and decasaccharide samples deemed to be pure by strong anion-exchange HPLC. In addition, subtle differences in the positioning of sulphate groups within heparan sulphate hexasaccharides were impossible to detect by strong anion-exchange HPLC. PAGE analysis on the other hand afforded excellent resolution of these structural isomers. The precise positioning of specific sulphate groups has been implicated in determining the specificity of heparan sulphate interactions and biological activities; hence, the purification of oligosaccharide species that differ in this way becomes an important issue. In this study, we have used strong anion-exchange HPLC and PAGE techniques to allow production of the homogeneous heparan sulphate oligosaccharide species that will be required for the detailed study of structure/activity relationships.
    Citation
    Combined strong anion-exchange HPLC and PAGE approach for the purification of heparan sulphate oligosaccharides. 2001, 354 (Pt 1):141-7 Biochem. J.
    Journal
    The Biochemical Journal
    URI
    http://hdl.handle.net/10541/85563
    PubMed ID
    11171089
    Type
    Article
    Language
    en
    ISSN
    0264-6021
    Collections
    All Paterson Institute for Cancer Research

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