A method for systematic purification from bovine plasma of six vitamin K-dependent coagulation factors: prothrombin, factor X, factor IX, protein S, protein C, and protein Z.
A systematic purification scheme is presented for the isolation of six vitamin K-dependent coagulation factors from bovine plasma in a functionally and biochemically pure state. The vitamin K-dependent proteins concentrated by the ordinary barium citrate adsorption were first separated into four fractions, fractions A, B, C, and D, by DEAE-Sephadex A-50 chromatography. From the pooled fraction A, protein S, factor IX, and prothrombin were purified by column chromatography on Blue-Sepharose CL-6B. Heparin-Sepharose chromatography of the pooled fraction B provided mainly pure factor IX, in addition to homogeneous prothrombin. A high degree of resolution of protein C and prothrombin from the pooled fraction C was obtained with a Blue-Sepharose column. This dye-ligand chromatographic procedure was also very effective for the separation of protein Z and factor X contained in the pooled fraction D. Thus, these preparative procedures allowed high recovery of milligram and gram quantities of six vitamin K-dependent proteins from 15 liters of plasma in only two chromatographic steps, except for protein S, which required three (the third step was rechromatography on Blue-Sepharose CL-6B).
Hashimoto
N
N
Morita
T
T
Iwanaga
S
S
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001779:Blood Coagulation Factors / Q000302:isolation & purification*; D001798:Blood Proteins / Q000302:isolation & purification; D002417:Cattle; D002845:Chromatography / Q000379:methods; D005164:Factor IX / Q000302:isolation & purification; D005170:Factor X / Q000302:isolation & purification; D006023:Glycoproteins / Q000302:isolation & purification; D008970:Molecular Weight; D011486:Protein C; D017293:Protein S; D011516:Prothrombin / Q000302:isolation & purification; D014812:Vitamin K
D000596:Amino Acids; D001779:Blood Coagulation Factors; D001798:Blood Proteins; D006023:Glycoproteins; D011486:Protein C; D017293:Protein S; C026080:plasma protein Z; D014812:Vitamin K; D011516:Prothrombin; D005164:Factor IX; D005170:Factor X