用昆虫细胞表达系统表达人组织激肽释放酶原

Expression of Human Tissue Prokallikrein in Insect Cells

目的:利用杆状病毒-昆虫细胞表达系统表达proHUK,系统优化表达条件。方法:用改进的方法对昆虫细胞进行了无血清悬浮适应培养,用ELISA、SDS-PAGE方法对各种条件下proHUK的表达量进行检测。结果:Sf-9、Hi-5细胞在血清减量速度为5%、1%,接种密度分别为2×106cells/mL1、×106cells/mL时能很快适应无血清悬浮培养。在病毒感染复度MOI为10,细胞接种密度为1×106cells/mL条件下培养96h后,proHUK的表达量最高可达30mg/L。结论:改进的方法使昆虫细胞能更快适应无血清悬浮生长条件,获得了高表达proHUK的方法,为其大规模制备奠定了基础。

Objective： Using baculovirus- insect cell system to express human tissue prokallikrein. To obtain the optimal expression conditions for the recombinant prokallikrein. Methods： A new method was developed to recuhure insect cells. The expression level was detected by EIJSA and SDS- PAGE. Results： The cells were reeuhured in medium without FBS, and further suspended in a spinner flask. The expression level of recombinant prokallikrein under MOI = Z10, cells inoculating density 1 × 10^6cells/mL eonditious was as high as 30mg/L. Conclusion： A new method to obtain high expression level prokalllkrein was developed.