GE Healthcare公司推出的AKTAxpress TWIN是一个整合的全自动蛋白纯化系统，用于多级纯化His和GST标记蛋白的小试研究。这个小型的高通量蛋白纯化系统整合了硬件、软件和介质，不需要复杂的操作程序。UNICORN软件帮助选择最佳纯化参数...GE Healthcare公司推出的AKTAxpress TWIN是一个整合的全自动蛋白纯化系统，用于多级纯化His和GST标记蛋白的小试研究。这个小型的高通量蛋白纯化系统整合了硬件、软件和介质，不需要复杂的操作程序。UNICORN软件帮助选择最佳纯化参数，自动将每一个纯化阶段中的峰值蛋白自动转移到下一个层析柱中，多级纯化出的蛋白纯度高于95％。展开更多
Enterovirus type 71（EV71） causes severe hand-foot-and-mouth disease （HFMD） resulting in hundreds of deaths of children every year; However, currently, there is no effective treatment for EV71. In this study, the E...Enterovirus type 71（EV71） causes severe hand-foot-and-mouth disease （HFMD） resulting in hundreds of deaths of children every year; However, currently, there is no effective treatment for EV71. In this study, the EV71 poly-protein （EV71-P1 protein） gene was processed and cloned into the eukaryotic expression vector pPIC9k and then expressed in Pichia pastoris strain GSll5. The EV71 P1 protein with a molecular weight of 100 kD was produced and secreted into the medium. The soluble EV71 P1 protein was purified by column chromatography with a recovery efficiency of 70%. The result of the immunological analysis showed that the EV71 P1 protein had excellent immunogenicity and could stimulate the production of EV71-VP1 IgG antibody in injected rabbits. We suggest that EV71-P1 protein is an ideal candidate for an EV71 vaccine to prevent EV71 infection.展开更多
基金Key Project of Science and Technology Department of Wuhan(zz2011-12)Key Project of the Educational Bureau of Wuhan(sz2011-13-10)+1 种基金Project of Science and Technology Department of Hubei(2010CDB04801)The State Key Laboratory program of Viral Genetic Engineering(2010KF10)
文摘Enterovirus type 71（EV71） causes severe hand-foot-and-mouth disease （HFMD） resulting in hundreds of deaths of children every year; However, currently, there is no effective treatment for EV71. In this study, the EV71 poly-protein （EV71-P1 protein） gene was processed and cloned into the eukaryotic expression vector pPIC9k and then expressed in Pichia pastoris strain GSll5. The EV71 P1 protein with a molecular weight of 100 kD was produced and secreted into the medium. The soluble EV71 P1 protein was purified by column chromatography with a recovery efficiency of 70%. The result of the immunological analysis showed that the EV71 P1 protein had excellent immunogenicity and could stimulate the production of EV71-VP1 IgG antibody in injected rabbits. We suggest that EV71-P1 protein is an ideal candidate for an EV71 vaccine to prevent EV71 infection.
