Purification and Properties of Guine Pig Antithrombin III

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Abstract
Guine pig antithrombin III has been purified from plasm by sequential heparin-Sepharose affinity chromatography, DE-52 cellu-lose chromatography, isoelectric focussing, and Sephadex G-100 gel filtration chromatography. The final product was homogeneous as judged by sodium dodecyl sulfate disc gel electrophoresis. Purifi-cation was 202-fold with yield of 41%. Antiproteinase activity of antithrombin III was determined by progressive inactivation of thrombin coagulant and amidolytic activity. Heparin cofactor activ-ity was demonstrated by immediate inactivation of thrombin by anti-thrombin III in the presence of minute quantities of heparin. It also could be demonstrated that thrombin inactivation by anti-thrombin III occurs by formation of bimolecular complex whose rate of formation is markedly enhanced by minute quantities of heparin.