A simultaneous purification of human prothrombin and factor IX

• Published in: Blood & Vessel Vol. 13; no. 1; pp. 63 - 71
• Main Authors: FUJIMURA, Yoshihiro, SAKAI, Toshiyuki, MATSUYAMA, Ikuko, MIKAMI, Sadaaki, YOSHIOKA, Akira, FUKUI, Hiromu
• Format: Journal Article
• Language: English
• Published: The Japanese Society on Thrombosis and Hemostasis 1982
• Subjects: DEAE-Sepharose CL-6B; factor IX; heparin-Sepharose 4B; preparative disc gel electrophoresis; prothrombin
• ISSN: 0386-9717; 1884-2372
• DOI: 10.2491/jjsth1970.13.63

Human factor IX has been purified from a commercial prothrombin complex concentrate (Proplex®) with a yield of 17%. The purified factor IX preparation had a specific activity of 171 units per mg protein. The purification procedure involves BaSO4 adsorption-elution, 40-70% saturated (NH4)2SO4 precipitation, DEAE-Sepharose CL-6B chromatography, heparin-Sepharose 4B chromatography, and preparative disc gel electrophoresis. The final product is homogeneous on an analytical polyacrylamide gel electrophoresis, and has an apparent molecular weight (mol. wt.) of 57, 000 which is not changed by chemical reduction. The monospecific antiserum to factor IX was raised against rabbits, and possessed an inhibitory activity equivalent to 35 Bethesda units per ml. In the course of factor IX purification, prothrombin (mol. wt. 72, 000) has also been purified (36% yield), and has been shown to have a specific activity of 8.6 units per mg protein.