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In recent years, we have presented several reports with respect to Chinese Yellow ( ocher originated from China ) as a new adsorbent for blood coagulation factors. Adsorbability of this ocher is strong for many coagulation factors in plasma including fibrinogen, while quite weak for antithrombins. After ocher adsorption treatment, AT HI activity of plasma in comperison with the value before adsorption is 86 - 96 % in clotting assay ( Biggs thrombin time methods ) and 96 - 100 % in immunologic activity (detected by SRID ). In this study, purification of plasma AT HI, we have used this ocher as the adsorbent in pretreatment of plasma together with heparin - sepharose affinity chromatography. As the actual procedure, 5 g of ocher was added to 100 ml of ACD plasma, and the mixture was subjected to adsorption with stirring at room temperature to obtain the ocher adsorbed plasma, crude AT HI fraction, in the supernatant after centrifugal separation. Highly purified AT HI with a molecular weight of approximately 67,000 ( determined by SDS-PAGE ) was obtained from the crude AT HE fraction by means of heparin - sepharose affinity chromatography. The antiserum from rabbits immunized with the purified AT HI showed a single fuse precipitin line against the starting plasma, ocher adsorbed plasma and AT HI fraction. However, it did not exhibit reaction with α2-macroglobulin and α1-antitrypsin. From the above results, it has been found that the ocher is an effective adsorbent in pretreatment of plasma for AT HI purification.

thrombosis and haemostasis

Purification Of Antithrombin Ul Using Chinese Yellow ( Ocher Originated From China ) As A New Adsorbent And Heparin - Sepharose Affinity Chromatography