Sinniger V, Tapon-Bretaudière J, Zhou F L, Bros A, Muller D, Jozefonvicz J, Fischer A M
Laboratoire d'Hématologie, C.H.U. Necker-Enfants Malades, Paris, France.
J Chromatogr. 1991 Feb 22;539(2):289-96. doi: 10.1016/s0021-9673(01)83937-6.
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.
硫酸皮肤素（DS）是一种糖胺聚糖，它能通过血浆抑制剂肝素辅因子II（HCII）特异性催化凝血酶抑制作用。将DS固定在二氧化硅基质上以研究其与HCII的相互作用。DS在预先涂有多糖的二氧化硅上以良好的产率进行固定，以中和带负电荷的硅醇基团。通过吸附等温线测量，固定化DS对HCII的亲和常数与可溶性DS获得的值一致。结合到二氧化硅基质上的DS也被测试作为从人血浆中纯化HCII的色谱支持物；确定了HCII吸附和解吸的最佳条件。洗脱得到的HCII产率良好（21%），且没有凝血酶的另一种主要血浆抑制剂抗凝血酶III的污染。
