title: Immobilization of dermatan sulphate on a silica matrix and its possible use as an affinity chromatography support for heparin cofactor II purification.
abstract: Dermatan sulphate (DS) is a glycosaminoglycan which catalyses specifically thrombin inhibition by a plasmatic inhibitor, Heparin cofactor II (HCII). DS was insolubilized on a silica matrix to study its interaction with HCII. The immobilization of DS was performed with a good yield on a silica previously coated with polysaccharides in order to neutralize the negatively charged silanol groups. The value of the affinity constant of insolubilized DS for HCII, measured by the adsorption isotherm, is consistent with the value obtained for soluble DS. The DS bound to the silica matrix was also tested as a chromatographic support for the purification of HCII from human plasma; the optimum conditions for HCII adsorption and desorption were determined. The eluted HCII was obtained with a good yield (21%) and with no contamination by antithrombin III, the other main plasmatic inhibitor of thrombin.
authors.affiliation: Laboratoire d'Hématologie, C.H.U. Necker-Enfants Malades, Paris, France.
authors.forename: V
authors.initials: V
authors.lastname: Sinniger
authors.forename: J
authors.initials: J
authors.lastname: Tapon-Bretaudière
authors.forename: F L
authors.initials: FL
authors.lastname: Zhou
authors.forename: A
authors.initials: A
authors.lastname: Bros
authors.forename: D
authors.initials: D
authors.lastname: Muller
authors.forename: J
authors.initials: J
authors.lastname: Jozefonvicz
authors.forename: A M
authors.initials: AM
authors.lastname: Fischer
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D003871:Dermatan Sulfate / Q000737:chemistry; D015844:Heparin Cofactor II / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D012822:Silicon Dioxide
chemical_list: D003871:Dermatan Sulfate; D012822:Silicon Dioxide; D015844:Heparin Cofactor II