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.
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February 2001
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Research Article|
February 08 2001
Combined strong anion-exchange HPLC and PAGE approach for the purification of heparan sulphate oligosaccharides
Romain R. VIVÈS;
Romain R. VIVÈS
*CRC Drug Development Group, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, U.K.
†The CRC and University of Manchester Department of Medical Oncology, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, U.K.
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Sarah GOODGER;
Sarah GOODGER
*CRC Drug Development Group, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, U.K.
†The CRC and University of Manchester Department of Medical Oncology, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, U.K.
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David A. PYE
David A. PYE
1
*CRC Drug Development Group, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, U.K.
†The CRC and University of Manchester Department of Medical Oncology, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, U.K.
1To whom correspondence should be addressed, at the Department of Medical Oncology (e-mail dpye@picr.man.ac.uk).
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Publisher: Portland Press Ltd
Received:
June 07 2000
Revision Received:
October 07 2000
Accepted:
October 27 2000
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 2001
2001
Biochem J (2001) 354 (1): 141–147.
Article history
Received:
June 07 2000
Revision Received:
October 07 2000
Accepted:
October 27 2000
Citation
Romain R. VIVÈS, Sarah GOODGER, David A. PYE; Combined strong anion-exchange HPLC and PAGE approach for the purification of heparan sulphate oligosaccharides. Biochem J 15 February 2001; 354 (1): 141–147. doi: https://doi.org/10.1042/bj3540141
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