人源TNF-α抗体基因的克隆与表达  

Cloning and expression of human TNF-α antibody gene

作　　者：田露芳[1] 赵小玲[2] 张克军[1] 杨志华[2] 

机构地区：[1]山西医科大学第一医院耳鼻喉科,山西太原030001 [2]军事医学科学院卫生学环境医学研究所应激医学研究室,天津300050

出　　处：《中国医药导报》2009年第21期9-11,共3页China Medical Herald

基　　金：国家自然科学基金(30700748);天津市科技攻关项目(06YFGPSH03600)

摘　　要：目的:在原核系统中高效表达抗TNF-α单克隆抗体VH/K21,以便进一步研究其生物学功能,预测临床应用前景。方法:以已筛选获得的TNF-α抗体基因VH/K21为模板进行PCR扩增,产物与其表达质粒进行双酶切,连接后克隆进入PET22b(+)/BL21(DE3)感受态中并在该原核系统中实现可溶性表达;对表达产物进行Western blotting、ELISA鉴定。结果:成功克隆与表达出可溶性的高效表达的人源TNF-α单克隆抗体。序列分析表明该克隆具有全新的人源TNF-α抗体基因,它可以特异识别人TNF-α。结论:人源TNF-α单克隆抗体的获得为进一步研制应用于临床的TNF-α人源抗体奠定了基础,也将为其他人源抗体的制备提供理论依据和技术基础。Objective： To express the TNF-α monoclonal antibody VH/K21 in the prokaryotic expression system, in order to further explore its biological functions and predict its clinical application prospect. Methods： The screened TNF-α antibody VH/K21 gene was amplified by PCR and cloned into the prokaryotic expression vector pET-22b（＋）/BL21（DE3）, and achieved soluble expression. The products were identified by Western blotting and ELISA. Results： The soluble and high expressed human TNF-α monoclonal antibody was successfully cloned and expressed. Sequence analysis suggested that this clone had fully new human TNF-α antibody gene, and it could specifically recognize human TNF-α. Conclusion： The obtain of human TNF-α monoclonal antibody lays a foundation for its clinical application, and would provide a theoretical and technical basis for the preparation of other human antibodies.

关 键 词：抗体 TNF-Α 变应性鼻炎 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]