METHOD FOR OBTAINING THROMBIN-LIKE COAGULATING ENZYME Russian patent published in 2005 - IPC

FIELD: medicine, laboratory diagnostics.

SUBSTANCE: the present innovation deals with blood testing based upon chromatographic purification of snake's venom, identification and mixing fractions that contain thrombin-like enzyme followed by isolating the target product due to chromatography upon Sepharose leveled with Tris-HCL buffer solution, moreover, snake's venom Agkistrodon halys in 0.05 M sodium chloride solution should undergo chromatography upon a column filled with Sephacryl S-300 HR, after identification and mixing fractions with thrombin-like enzyme they should be concentrated due to ultrafiltration at selecting against molecular weight of 10 Kd, the obtained solution should undergo chromatography upon a column with DEAE Sepharose FF leveled with acetate buffer solution to select and mix fractions with thrombin-like enzyme and repeatedly concentrate due to ultrafiltration, then concentrate should pass chromatography upon a column with SP Sepharose FF gel, fractions that contain the target product should be combined, diluted with distilled water up to activity of 12-15 sec in thrombin time test, then one should add saccharose, sodium azide up to 0.01% to be poured into vials and freeze dried. The method provides broadened quantity of reagents for laboratory diagnostics of disorders in the system of hemostasis. Fraction isolated out of snake's venom Agkistrodon Halys (the Russian Federation) is not inferior to that of snake's venom Bothrops atrox.

EFFECT: higher efficiency.

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