Toulon P, Chadeuf G, Aiach M
Laboratoire d'Hémostase, Hôpital Broussais, Paris, France.
J Chromatogr. 1991 Feb 22;539(2):493-500. doi: 10.1016/s0021-9673(01)83959-5.
Heparin cofactor II (HCII) is an inhibitor of thrombin in human plasma whose activity is enhanced by heparin and dermatan sulphate. HCII was purified to homogeneity from normal human plasma with an overall yield of 7.5%. After treatment with barium chloride, precipitation with 50% saturated ammonium sulphate and dialysis of the resuspended precipitate against 0.02 M Tris-HCl (pH 7.4), the sample was chromatographed on a heparin-Sepharose CL 6B affinity column, DEAE-Sepharose CL 6B ion-exchange gel and an AcA 34 gel permeation column. For the final steps, a high-performance liquid chromatographic system was used which included ion-exchange chromatography on a Mono-Q column and gel permeation using a Superose column. The purified protein was homogeneous by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The specific activity of purified HCII was 12.2 U/mg. The HCII activity was evaluated as antithrombin dermatan sulphate cofactor activity. A specific antiserum against HCII was raised in the rabbit.
肝素辅因子II（HCII）是人体血浆中凝血酶的一种抑制剂，其活性可被肝素和硫酸皮肤素增强。HCII从正常人血浆中纯化至均一，总产率为7.5%。经氯化钡处理、50%饱和硫酸铵沉淀以及将重悬的沉淀对0.02M Tris-HCl（pH 7.4）进行透析后，样品在肝素-琼脂糖CL 6B亲和柱、DEAE-琼脂糖CL 6B离子交换凝胶和AcA 34凝胶渗透柱上进行层析。对于最后步骤，使用了高效液相色谱系统，其中包括在Mono-Q柱上进行离子交换色谱以及使用Superose柱进行凝胶渗透。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，纯化后的蛋白质是均一的。纯化的HCII的比活性为12.2 U/mg。HCII活性作为抗凝血酶硫酸皮肤素辅因子活性进行评估。在兔体内制备了针对HCII的特异性抗血清。
