摘要
目的:纯化制备rhIL-21,并分析其免疫学活性。方法:在含重组质粒pGEX4T-2/IL-21的大肠杆菌DH5α中,经IPTG诱导表达出GST-融合蛋白,蛋白经过一系列的纯化,SDS-PAGE电泳鉴定蛋白纯度。用MTT法研究rhIL-21对T细胞增殖、NK细胞杀伤活性的影响。结果:表达出了可溶性融合蛋白,纯化后的rhIL-21纯度达95%。rhIL-21能促进T细胞增殖,能增强NK细胞杀伤活性。结论:成功制备了rhIL-21,其在体外具有一定的免疫学活性,为其大量制备和体内的生物学活性研究奠定了基础。
Objective:To prepare and purify recombinant human interleukin 21(rhIL-21),and to study its immunologicd activity in vitro.Methods:The E.coli DH5α containing recombinant plasmid pGEX4T-2/IL-21 was induced to express protein by IPTG.Supernatants of lysate were purified by affinity chromatography,thrombin digestion and cation chromatography sequentially.The purified products were analyzed by SDS-PAGE.The effect of rhIL-21 on proliferation of T lymphocyte and cytotoxicity of NK cells was studied by using MTT method.Results:The target protein was soluble in supernatants of the germ culture.Purity of rhIL-21 was 95% after the procedures for purification.rhIL-21 augmented proliferation of T lymphocytes and cytotoxicity of NK cells in vitro.Conclusion:rhIL-21 was successfully prepared in small scale and purified.It laid a foundation of study on biological activity in vivo,the procedure of pilot and large-scale preparations of rhIL-21.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2009年第1期72-75,共4页
Chinese Journal of Immunology
基金
山东省自然科学基金(E99C02)
青岛市科技局计划项目(04-2-J2-95)资助
关键词
人白细胞介素21
大肠杆菌
表达
纯化
活性
Human interleukin 21
Escherichia coli
Expression
Purification
Activity