药品评价
藥品評價
약품평개
DRUG REEVALUATION
2014年
24期
29-31
,共3页
五步蛇毒%凝血酶样酶%凝胶过滤色谱%阴离子交换色谱
五步蛇毒%凝血酶樣酶%凝膠過濾色譜%陰離子交換色譜
오보사독%응혈매양매%응효과려색보%음리자교환색보
Agkistrodon Acutusvenom%Thrombin-like Enzyme%Gel Filtration Chromatography%DEAE-Sepharose Fast Flow Ion-exchange
目的：改进分离工艺，找到液相色谱分离纯化五步蛇毒中凝血酶样酶的最佳方案。方法：五步蛇毒依次经Superdex G75凝胶过滤、DEAE-Sephrose F.F阴离子交换和Superdex G30凝胶过滤，通过改变洗脱液pH值、流速、Tris浓度、盐浓度及上样量来考察对分离纯化五步蛇毒中凝血酶样酶的影响。结果：Superdex 75凝胶过滤色谱分离纯化受洗脱液流速、Tris浓度、盐浓度、pH值、上样量的影响；DEAE-Sephrose F.F阴离子交换色谱受盐浓度的影响；Superdex G30凝胶过滤色谱受上样量的影响。三步分离纯化后获得一种五步蛇毒凝血酶样酶，分子量约23kD。结论：依次经Superdex G75、DEAE-Sephrose F.F和Superdex G30凝胶，通过改变洗脱液流速、Tris浓度、盐浓度、pH值、上样量等能得到最佳分离条件，最终可获得五步蛇毒凝血酶样酶的纯品。
目的：改進分離工藝，找到液相色譜分離純化五步蛇毒中凝血酶樣酶的最佳方案。方法：五步蛇毒依次經Superdex G75凝膠過濾、DEAE-Sephrose F.F陰離子交換和Superdex G30凝膠過濾，通過改變洗脫液pH值、流速、Tris濃度、鹽濃度及上樣量來攷察對分離純化五步蛇毒中凝血酶樣酶的影響。結果：Superdex 75凝膠過濾色譜分離純化受洗脫液流速、Tris濃度、鹽濃度、pH值、上樣量的影響；DEAE-Sephrose F.F陰離子交換色譜受鹽濃度的影響；Superdex G30凝膠過濾色譜受上樣量的影響。三步分離純化後穫得一種五步蛇毒凝血酶樣酶，分子量約23kD。結論：依次經Superdex G75、DEAE-Sephrose F.F和Superdex G30凝膠，通過改變洗脫液流速、Tris濃度、鹽濃度、pH值、上樣量等能得到最佳分離條件，最終可穫得五步蛇毒凝血酶樣酶的純品。
목적：개진분리공예，조도액상색보분리순화오보사독중응혈매양매적최가방안。방법：오보사독의차경Superdex G75응효과려、DEAE-Sephrose F.F음리자교환화Superdex G30응효과려，통과개변세탈액pH치、류속、Tris농도、염농도급상양량래고찰대분리순화오보사독중응혈매양매적영향。결과：Superdex 75응효과려색보분리순화수세탈액류속、Tris농도、염농도、pH치、상양량적영향；DEAE-Sephrose F.F음리자교환색보수염농도적영향；Superdex G30응효과려색보수상양량적영향。삼보분리순화후획득일충오보사독응혈매양매，분자량약23kD。결론：의차경Superdex G75、DEAE-Sephrose F.F화Superdex G30응효，통과개변세탈액류속、Tris농도、염농도、pH치、상양량등능득도최가분리조건，최종가획득오보사독응혈매양매적순품。
Objective:This study was designed to separate and purify the component of thrombin-like enzyme from Agkistrodon acutus venom by the new technology. Methods:Proteins were separated and purified by Superdex 75, DEAE-Sepharose Fast Flow and Superdex 30, with the changing of the flow rate, pH value, concentrations of Tris and NaCl, and the sample volume. Results:The flow rate, pH value, concentrations of Tris and NaCl, as well as the sample volume affected Superdex 75' s separation effect. The concentration of NaCl influences DEAE-Sepharose Fast Flow' s separation. The sample volume influences Superdex 30' s separation effect. The thrombin-like enzyme protein was purified from Agkistrodon acutus venom with a molecular weight around 25 kD after the three-step separation. Conclusion:By changing the eluent flow rate, the concentrations of Tris and NaCl, pH value, volume of sample, the best separation conditions and purified thrombin-like enzyme from Agkistrodon acutus venom can be obtained.
