Objective To provide a new purification process for snake venom thrombin-like enzyme.Methods The snake venom was pretreated and purified by Benzamidine Sepharose 6B affinity chromatography,SP Sepharose High Performance cation exchange chromatography and Phenyl Sepharose 6 Fast Flow hydrophobic chromatography,and finally the venom thrombin-like enzyme was obtained.The enzyme was detected by coagulometer for activity,reduced SDS-PAGE for molecular weight,non-reduced SDS-PAGE,capillary electrophoresis and high-performance liquid chromatography for purity.And the quality comparison was studied between the enzyme purified by the new process and the existing patented process.Results There was no significant difference in molecular weight between the two kinds of purification process,but the enzyme purified by the new process had the higher specific activity,purity and better process stability than that by the existing patented process.Conclusion Compared with the existing patented process,the new process simplifies the process steps,shortens the purification cycle,reduces the risk of exposure to the process environment.The high-purity thrombin-like enzymes with good process stability and quality consistency could be obtained,which is conducive to clinical drug safety.
关键词
蛇毒类凝血酶/纯化/新方法
Key words
snake venom thrombin-like enzyme/purification/new process
维普
万方数据