HPV58型E2蛋白表达纯化与多克隆抗体制备  

Expression and purification of HPV58 E2 protein and preparation of polyclonal antibodies against HPV58 E2 protein

作　　者：姜文婷[1] 程一航[1] 唐海平[2] 谢毅[1] 连文博 詹建民[2] 徐万祥[2] 顾少华[1] 

机构地区：[1]复旦大学生命科学学院,遗传工程国家重点实验室,上海市工业菌株工程技术研究中心,上海200438 [2]上海市计划生育科学研究所,国家人口和计划生育委员会重点实验室,上海200032

出　　处：《生殖与避孕》2015年第10期674-679,共6页Reproduction and Contraception

基　　金：上海市科委资助项目,项目号:13DZ2252000

摘　　要：目的:表达纯化重组人乳头瘤病毒(human papillomavirus,HPV)58型E2蛋白,制备多克隆抗体。方法:构建重组质粒p ET-28b-E2,转化至BL21(DE3)p Lys S中诱导表达,包涵体洗涤后经镍柱亲和层析分离得到纯化目的蛋白。用纯化的重组E2蛋白免疫新西兰白兔,制备兔抗HPV58型E2蛋白多克隆抗体,ELISA分析多克隆抗体的效价,免疫印迹检测抗体的特异性。结果:表达纯化了重组HPV58型E2蛋白,制备了高滴度和高特异性的多克隆抗体。结论:制备的多克隆抗体可用于对HPV58型E2蛋白进行精细B细胞线性表位鉴定。Objective： To express and purify recombinant E2 protein of human papillomavirus（HPV） 58 and to obtain polyclonal antibodies against HPV58 E2 protein. Methods： The constructed recombinant plasmid p ET-28b-E2 was transformed into BL21（DE3） p Lys S and recombinant E2 protein expression was induced. Recombinant E2 protein was purified by nickel column affinity chromatography after inclusion body being washed.Immunizing New Zealand white rabbits with purified recombinant E2 protein, rabbit anti-HPV58 E2 polyclonal antibodies were prepared. After then, the polyclonal antibodies titer were analyzed by ELISA and the polyclonal antibodies specificity to recombinant E2 protein was detected through Western blotting. Results： We successfully expressed and purified recombinant HPV58 E2 protein and prepared the polyclonal antibodies of high titer and high specificity to recombinant E2 protein. Conclusion： The polyclonal antibodies can be used to further identify exquisite linear B-cell epitopes of HPV58 E2 protein.

关 键 词：HPV58型E2蛋白 蛋白表达 包涵体纯化 抗体制备 

分 类 号：R392-33[医药卫生—免疫学]