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重组人白细胞介素17的纯化及生物活性鉴定

Purification and bioactiveity detection of recombinant human interleukin 17

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【Author】 ZHANG Fushan 1),LI Rongxiu 2),LIU Zhongbin 3),QIN Xia 3) 1)Clinical Laboratory,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052 2)School of Life Science and Technology,Shanghai Jiaotong University, Shanghai 200030 3)Department of Microbiology and Immunology,Medical College,Tongji University,Shanghai 200331

【机构】郑州大学第一附属医院检验科；上海交通大学生命科学技术学院；同济大学医学院微生物学与免疫学教研室；同济大学医学院微生物学与免疫学教研室郑州450052；上海200030；上海200331；

【摘要】目的:探讨重组人白细胞介素17的复性与纯化方法,并进行活性鉴定。方法:表达重组人白细胞介素17(rhIL17)的大肠杆菌经破碎、包涵体洗涤、裂解提取、复性、SephacrylS100层析柱纯化等方法,获得rhIL17;通过ELISA方法检测IL6的含量,间接反映rhIL17的生物学活性。结果:经纯化得到rhIL17,其相对分子质量为159000,蛋白纯度为97.2%,能够刺激人胚肺成纤维细胞MRC5产生IL6,呈浓度依赖关系。结论:通过本方法复性与纯化获得高纯度、有生物学活性的rhIL17。更多还原

【Abstract】 Aim: To study the renaturation,purification, and bioactivity detection of recombinant human interleukin-17 (rhIL-17) . Methods: Recombinant rhIL-17 expression in E.coli was isolated and purified by washing and solubilization of inclusion body,renaturation, chromatography separation and dialysis.The bioactivity was detected by determining the levels of IL-6 after its stimulating MRC -5 cells.Results: rhIL-17 was obtained with a purity of 97.2%.Molecular weight of rhIL-17 was 159 000,and bioactivity experiment showed that rhIL-17 stimulated MRC-5 cells to secrete IL-6 in a dose-dependent manner.Conclusion: rhIL-17 with high purity and bioactivity could be obtained with this method.更多还原

【关键词】人白细胞介素17；纯化；生物学活性；
【Key words】 human interleukin 17； purification； bioactivity；

【文献出处】郑州大学学报(医学版) ,Journal of Zhengzhou University(Science Medical) , 编辑部邮箱 ,2005年06期

【分类号】R392
【下载频次】137

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