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

2.50
Hdl Handle:
http://hdl.handle.net/10541/85563
Title:
Combined strong anion-exchange HPLC and PAGE approach for the purification of heparan sulphate oligosaccharides.
Authors:
Vivès, Romain R; Goodger, Sarah J; Pye, David A
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.
Affiliation:
CRC Drug Development Group, and University of Manchester Department of Medical Oncology, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, UK.
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
Issue Date:
15-Feb-2001
URI:
http://hdl.handle.net/10541/85563
PubMed ID:
11171089
Type:
Article
Language:
en
ISSN:
0264-6021
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorVivès, Romain Ren
dc.contributor.authorGoodger, Sarah Jen
dc.contributor.authorPye, David Aen
dc.date.accessioned2009-11-06T14:47:31Z-
dc.date.available2009-11-06T14:47:31Z-
dc.date.issued2001-02-15-
dc.identifier.citationCombined strong anion-exchange HPLC and PAGE approach for the purification of heparan sulphate oligosaccharides. 2001, 354 (Pt 1):141-7 Biochem. J.en
dc.identifier.issn0264-6021-
dc.identifier.pmid11171089-
dc.identifier.urihttp://hdl.handle.net/10541/85563-
dc.description.abstractHeparan 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.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshChromatography, High Pressure Liquid-
dc.subject.meshChromatography, Ion Exchange-
dc.subject.meshElectrophoresis, Polyacrylamide Gel-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshSwine-
dc.titleCombined strong anion-exchange HPLC and PAGE approach for the purification of heparan sulphate oligosaccharides.en
dc.typeArticleen
dc.contributor.departmentCRC Drug Development Group, and University of Manchester Department of Medical Oncology, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, UK.en
dc.identifier.journalThe Biochemical Journalen

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