马血凝血酶原纯化方法的比较研究

Comparative Study on Purification Methods of Blood Coagulation Enzymes in Horse Blood

文章利用等电点法提取马凝血酶原并经过超滤膜和大孔阴离子交换树脂纯化,比较两种纯化方法的效果。方法:等电点法提取马凝血酶原液后,用最终浓度为0.1mol/L的Ca Cl2于30℃水浴中激活2.5h测酶活力。超滤膜纯化参数:30k Da再生纤维素膜,料液5℃,流速35.67m/s,浓缩2倍激活后测凝血酶活力。大孔树脂纯化法参数:上样流速0.5m L/min,以1%-100%的洗脱剂梯度洗脱,最佳比例为60%Na Cl,激活后测酶活力。结果:粗酶活力为119U/m L;超滤后为249U/m L;大孔树脂纯化后酶活力为301U/m L。结果表明大孔树脂的纯化效果优于超滤膜的纯化效果。

Use of isoelectric point extraction horse prothrombin and after ultrafiltration membrane and macroporous anion exchange resin purification,the effect of two purification methods. Methods：Extraction of the isoelectric point of the horse thrombin stock solution,0.1 mol/L Ca Cl2 activation of the enzyme activity measured at 2.5 h30 ℃ water bath with final concentration. UF purification parameters：30 k Da regenerated cellulose membrane,liquid 5 ℃,the flow rate of 35.67 m/s,twice as concentrated measuring thrombin activation energy. Macroporous method parameters：sample flow rate 0.5 m L/min,with 1% to 100% gradient of eluent,the optimum ratio of 60%Na Cl,the enzyme activity measured after activation. Results：The crude enzyme activity was 119 U/m L; ultrafiltration is 249 U/m L; enzyme activity after macroporous resin purification of 301 U/m L. The results show that the effect of macroporous resin purification superior ultrafiltration membrane purification effect.