重组人白细胞介素-18的纯化、复性及其活性检测  

Purification,Renaturation and Biological Activity of Recombinant Human Interleukin-18

作　　者：陈文芳[1] 张利宁[1] 宋磊 刘志亭 刘应军 

机构地区：[1]山东大学医学院免疫学研究所,济南250012 [2]山东泉港药业有限公司,济南250011

出　　处：《中国生物制品学杂志》2005年第3期238-240,共3页Chinese Journal of Biologicals

基　　金：山东省科学技术发展计划项目(021100103)(本项目属山东泉港药业有限公司).

摘　　要：目的纯化和复性rhIL18,并测定其生物学活性。方法采用提取包涵体、分子筛层析及离子交换层析的方法纯化rhIL18,经稀释法复性,用IL18诱导PBMC产生IFNγ,ELISA法检测上清中IFNγ的水平。结果rhIL18纯度大于95%,复性回收率达30%以上,获得的产品可诱导PBMC产生大量的IFNγ,且呈剂量依赖关系。结论该纯化、复性方法为大量生产rhIL18打下了基础。Objective To purify and re-naturalize recombinant human interleukin-18(rhIL-18) and determine its biological activity.Methods Extract inclusion body of rhIL-18,purify by molecule sieve and ion-exchange chromatography and re-naturalize by dilution.The re-naturalized rhIL-18 was used for induction of secretion of IFN-γ by PBMC,and the IFN-γ level was determined by ELISA.Results The purity and recovery rate of rhIL-18 were more than 95% and more than 30% respectively.The re-naturalized rhIL-18 showed significant and dose-dependent activity in the induction of IFN-γ in PBMC.Conclusion The procedures for purification and re-naturalization laid a foundation of large-scale production of rhIL-18.

关 键 词：白细胞介素-18 复性 纯化 活性检测 重组人 IFN-γ 离子交换层析 ELISA法 PBMC 生物学活性 IL-18 依赖关系 大量生产 包涵体 分子筛 稀释法 回收率 诱导 

分 类 号：Q786[生物学—分子生物学] S852.4[农业科学—基础兽医学]