Rapid purification of factor IX, factor X and prothrombin by immunoaffinity and ion exchange chromatography.
This study describes a rapid purification of factor IX, factor X and prothrombin by immunoaffinity and ion exchange chromatography. Human factor IX was purified from plasma in 34% yield using barium sulfate adsorption and immunoaffinity purification. The specific clotting activity of purified factor IX was 269 units/mg, with an apparent molecular weight of 57,000 in the presence of sodium dodecyl sulfate on polyacrylamide gels. The immunodepleted, factor-IX deficient plasma was chromatographed on a dextran sulfate agarose column which resolved prothrombin and factor X in highly purified states and with approximately 50% yield. The specific activities of prothrombin and factor X obtained by this procedure were 24 units/mg and 147 units/mg respectively. Both proteins isolated by this method showed a single component on SDS gel electrophoresis and the molecular weights of intact prothrombin and factor X were 72,000 and 67,000 respectively.
Department of Medicine, Temple University School of Medicine, Philadelphia, Pennsylvania 19140.
S S
SS
Ahmad
R
R
Rawala-Sheikh
A R
AR
Thompson
P N
PN
Walsh
D000645:Ammonium Sulfate; D000863:Antacids; D000911:Antibodies, Monoclonal; D001780:Blood Coagulation Tests; D001798:Blood Proteins / Q000032:analysis; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D002951:Citrates; D019343:Citric Acid; D005164:Factor IX / Q000302:isolation & purification*; D005170:Factor X / Q000302:isolation & purification*; D006801:Humans; D011516:Prothrombin / Q000302:isolation & purification*
D000863:Antacids; D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D002951:Citrates; D019343:Citric Acid; D011516:Prothrombin; D005164:Factor IX; D005170:Factor X; D000645:Ammonium Sulfate