Abstract

The optimal conditions for immobilizing heparin through its terminal formyl group were investigated. When Amino Sepharose (1 g) was suspended in 1 ml of phosphate buffer (pH 7) containing 30 mg of heparin and 3 mg of sodium cyanoborohydride, with shaking at room temperature, the maximum immobilization of heparin (10 mg of heparin per gram of wet gel) was reached within 2 days. The heparin sepharose thus obtained was stable: no significant loss of the heparin content was observed after storage for 4 months at 4°C. Heparin was also immobilized by the same method with amino TSK gel G5000PW instead of Amino Sepharose 4B and was successfully applied to the high-performance liquid affinity chromatography of fibronectin and thrombin.

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