从Cohn氏法组分Ⅲ沉淀纯化人凝血因子Ⅶ

Isolation and purification of human coagulation factor Ⅶ from Cohn fraction Ⅲ paste

目的探索从Cohn氏法组分Ⅲ(组分Ⅲ)沉淀中纯化人凝血因子Ⅶ(FⅦ)的方法并对纯化产物进行鉴定。方法将组分Ⅲ沉淀溶解,经柠檬酸钡吸附、洗脱、硫酸铵沉淀、2次DEAE-Sepharose Fast Flow离子交换层析和Sephadex G-100凝胶过滤纯化人FⅦ。结果从400g组分Ⅲ沉淀中最终得到10.1mg纯化的FⅦ,活性为1775.8U/mg,FⅦ被浓缩了近6000倍,总活性回收率为17.6%,SDS-PAGE电泳鉴定只有1条主条带。结论组分Ⅲ沉淀中有很高FⅧ活性;建立了从组分Ⅲ沉淀中纯化人FⅦ的方法。

Objective To isolate and purify human coagulation factor Ⅶ from Cohn fraction m precipitate. Methods The purification procedure of human factor Ⅶ from Cohn fraction Ⅲ precipitate involves dissolving fraction Ⅲ, absorbing factor Ⅶ onto barium citrate and during, ammonium sulfate fractionation and DEAE-Sepharose Fast Flow ion exchange chromatography, and Sephadex G-100 gel filtration chromatography. Results 10. 1mg purified FⅦ was obtained from 400g Cohn fraction Ⅲ precipitate. The purified FⅦ has a specific clotting activity of 1775.8U/mg and the overall yield of FⅦ specific clotting activity is 17.6 % of the starting material. The purity of FⅦ was judged by SDS-PAGE and there was only one protein band on the gel. Conclusion The procedure of purifying Ⅶ from Cohn fraction Ⅲ precipitate is established with satisfactory purity.