Isolation and purification of human coagulation factor Ⅶ from Cohn fraction Ⅲ paste / 中国输血杂志

ABSTRACT

Objective To isolate and purify human coagulation factor Ⅶ from Cohn fraction Ⅲ 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 eluting,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 Ⅲprecipitate is established with satisfactory purity.