[Isolation of human thrombin using affinity chromatography on gramicidin C-silochrome C 80].
Thrombin purification is conducted by biospecific chromatography on gramicidin C-silochrome C 80. Preparations possessing the fibrinogen-coagulating activity of 2500-3200 NIH units per 1 mg of protein and containing 98% of active sites are obtained. Data obtained from electrophoresis in PAAG with the presence of DS-Na show the alpha-thrombin content to be 96%; the admixture of beta-thrombin possessing no coagulating activity does not exceed 4%. The kinetic constants are presented for thrombin hydrolysis of tosyl-L-arginine methyl ester (TAME), benzoyl-L-arginine ethyl ester (BAEE) and chromogenic substrate S-2238. The addition of isopropanol increases sharply the stability of thrombin when storing it in the aqueous-salt solutions.
D002846:Chromatography, Affinity; D006096:Gramicidin; D006801:Humans; D008024:Ligands; D058428:Silica Gel; D012822:Silicon Dioxide; D013917:Thrombin / Q000302:isolation & purification*
D008024:Ligands; D006096:Gramicidin; D058428:Silica Gel; D012822:Silicon Dioxide; D013917:Thrombin
Poluchenie trombina cheloveka s pomoshch'iu affinnoĭ khromatografii na gramitsidin C-silokhrome C 80.