一种新的灵长类IL-4基因表达产物纯化方法  

An Innovative Method for the Purification of Recombinant IL-4 from Macaca mulatta

作　　者：袁竹青[1] 李剑[1] 汤云霞[2] 吴忠道[3] 余新炳[3] 

机构地区：[1]广州医学院病原生物学教研室,广州510182 [2]广州医学院第一附属医院,广州510120 [3]中山大学中山医学院寄生虫学教研室,广州510080

出　　处：《热带医学杂志》2010年第11期1277-1280,共4页Journal of Tropical Medicine

摘　　要：目的研究一种能保持重组蛋白活性的新的蛋白纯化方法。方法 IPTG诱导表达重组质粒pET-30a(+)-IL-4,取可溶性部分进行Ni-NTA agarose非变性方法纯化,随后逆向利用Biologic Duoflow Chromatography系统阴离子交换柱进一步分纯,并用Western Blot鉴定所得纯化蛋白。结果重组质粒pET-30a(+)-IL-4有部分可溶性表达,经Ni-NTA agarose非变性的方法纯化后仍有部分杂蛋白的存在,利用Biologic Duoflow Chromatography系统阴离子交换柱进一步分纯后得到高度纯化的重组蛋白并经Western Blot鉴定为目的蛋白。结论利用创新方法经两个纯化步骤得到了有活性的目的蛋白。Objective Develop an innovative method to purify biologically active recombinant protein.Methods Recombinant IL-4 expression plasmid pET-30a（＋） was induced by IPTG and Ni-NTA agarose was used for the purification of recombinant protein.Biologic Duoflow Chromatography system was used for further purification.The purified protein was identified by Western Blot.Results Soluble recombinant IL-4 was purified using Ni-NTA agarose and Biologic Duoflow Chromatography.The protein was confirmed by Western Blot.Conclusion Active recombinant IL-4 can be purified by these two steps purification method.

关 键 词：IL-4 蛋白纯化 Chromatography系统 

分 类 号：R284.2[医药卫生—中药学]