Purification Anticoagulation Protein From Human Placenta And Its Effects On Human Plasma Coagulation

Objective: To purify anticoagulation protein from human placental and study the biochemical charaterization and anticoagulation effect in vitro. Methods: The placental anticoagulation protein was purified from homogenated human placental by ammonium sulfate precipitation and column chromatography on DEAE Sepharose CL-6B, Sephadex G-75 and Heparin Sepharose CL-6B.Using SDS-PAGE calcuated its moelcular weight and them estimated isoelectric point by the isoelectric focusing electrophoresis; and assayed anticoagulant activities in vitro by Acitivated partia thromboplastin time (APTT), Prothrombin time (PT), Thrombin time (TT) , The content of fibrinogen(FIB) and Blood clotting factor VIII and IX. Result: The purified protein showed a single band by SDS-PAGE with a moelcular weight of 34.9KD .The isoelectric point was pH4.9 by The isoelectric focusing electrophoresis. This protein prolonged acitivated partial thromboplastin time and thrombin time in vitro,but had no effect on prothrombin time and fibrinogen. It also had inhibition on factor VIIIand factor IX. The inhibition of the protein on factor VIII was more stronger than factor IX and its effect on APTT was more stronger with increasing its concentration. Conclution: The placentalanticoagulation protein has strong anticoagulation activity, it may be inhibit the coagulation process by the entogenous way.