摘要
以新鲜猪血为原料 ,采用离心、激活、沉淀等技术提取出粗凝血酶 ,利用化学方法将分子量范围在 6KD左右的肝素连接到经溴化氰活化的 Sepharose CL- 6B树脂上 ,制成亲和树脂 ;采用亲和方法 ,从粗酶中纯化得到猪凝血酶。活性测定表明 ,在保持酶的底物专一性的同时 ,凝血酶的比活上升了 32 .5倍左右。计算总活力回收率约 5 0 .6%。同时对肝素亲和层析柱的制备以及洗脱方法进行了探讨。
In this article the affinity chromatography which served as a fast purification technique for the thrombin was studied. A rude thrombin was made from fresh blood of pigs by means of centrifugation, activation and precipitation etc.. The affinity chromatography was made by immobilizing the heparin with molecular masses about 6kD to Sepharose CL-6B which was activated by HBr. Then the thrombin was purified by the affinity chromatography. According to the activity test, which guaranteed the specific to the substances, the specific activity of the enzyme was raised to 32.5 times and the total enzyme activity is 50.6% of original activity. Also the preparation of the affinity chromatography and the elution methods was discussed.
出处
《青岛科技大学学报（自然科学版）》
CAS
2003年第3期225-228,共4页
Journal of Qingdao University of Science and Technology:Natural Science Edition
基金
青岛市海洋酶工程重点实验科研基金资助 ( 2 0 0 2 -0 0 18)
