Purification of a Highly Potent Heparin-Like Anticoagulant from Healthy Human Plasma
A highly potent-heparin-like anticoagulant (accelerator) has been purified from citrated healthy human plasma. After heat defibrination and BaSO4. treatment on the plasma, the accelerator was adsorbed onto a DEAE-cellulose column and elution was achieved using a high ionic strength linear buffered salt gradient. The eluted accelerator was further purified by polyacrylamide gel electrophoresis. The purified accelerator can accelerate the inhibition of thrombin by antithrombin III and has a specific activity of 226 heparin units/mg of carbohydrate. The accelerator contains a small amount of protein-like material. The amount of accelerator present in healthy human blood is extremely small and can only be first detected in the concentrate after the DEAE-cellu-lose chromatography. A mere 0.5 mg of the purified accelerator is obtained from 100 ml of human plasma. Concomitant with this investigation, a second heparin-like substance also has been purified but has very low anticoagulant activity in terms of heparin units. The naturally occurring accelerator may function as heparin in the circulating blood and its level in blood may have a clinical significance in thrombotic vascular disease. Further work on its physical and chemical properties is now in progress.