抗trop2人源抗体fab片段原核表达的优化 - 南京医科大学.doc

全人源抗狂犬病病毒scFv-Fc融合抗体的制备及活性分析 高畅1 张倩倩1 熊四平1 唐小军1 仇镇宁3 冯振卿1,3§* 朱进2,3* （1南京医科大学病理学系，南京210029; 2 南京军区军事医学研究所，南京210002; 3 卫生部抗体技术重点实验室， 南京210029） 【摘要】 【关键词】Preparation and identification of the binding activity of a whole human scFv-Fc fusion antibody against rabies virus GAO Chang1, ZHANG Qian-qian1 , XIONG Siping1 ,TANG Xiao-jun 1 ,QIU Zhen-ning 3 FENG Zhen-qing 1§ , ZHU Jin 1,2 (1 Department of Pathology,, Nanjing Medical University, Nanjing 210029; 2 Huadong Medical Institute of Biotechniques, Nanjing 210002; 3 Key Laboratory of Antibody Technique of Ministry of Health, Nanjing Medical University, , Nanjing 210029;) [Abstract] Objective: To prepare a human scFv-Fc fusion antibody and to analyze its binding activity. Methods: The human scFv-Fc-pColdⅡprokaryotic expression vector was constructed. After optimizing prokaryotic expression conditions at different concentration of IPTG, the human scFv-Fc antibody was expressed and purified.Purified humanized scFv-Fc antibody was confirmed its binding activity by binding to purified inactivated rabies virus. Results: After sequence analysis, the human scFv-Fc-pColdⅡprokaryotic expression vector was successfully established. Through transforming E.coli.BL21(DE3), human scFv-Fc fusion antibody was expressed by optimized induction of IPTG at concentration of 0.5mmol/L .By His-trap column purification, the human scFv-Fc fusion antibody was purified with excellent binding activity even in 512 times diluted concentration. Conclusions: We have successfully established a whole human anti-rabies virus scFv-Fc fusion antibody with excellent binding activity. This fusion antibody can lay a foundation for developing novel antibody-targeted drugs of rabies prophylaxis after exposure. [Key words] rabies virus; whole human; scFv-Fc antibody §[基金项目]国家自然科学基金资助 *通讯作者，E-mail:冯振卿 fengzhenqing@njmu.edu.cn； 朱进 zhujin1968@njmu.edu.cn 狂犬病是由狂犬病病毒引起的人畜共患病[1]，我国是受狂犬病危害最为严重的国家之一，仅次于印度，居全球第二位，狂犬病一直位于我国各类传染病报告死亡数的前三位。根据最新WHO推荐