抗人转铁蛋白单链抗体的研制及部分性能研究  

Preparation and partial characterization of ScFv antibody to human transferrin

作　　者：李兆隆[1] 郭红玲[1] 常彩琴[1] 涂政[1] 肖卫民[2] 王明鑫[1] 王俭[1] 

机构地区：[1]公安部物证鉴定中心,北京100038 [2]北京大学生命科学学院,北京100871

出　　处：《中国法医学杂志》2001年第S1期12-16,共5页Chinese Journal of Forensic Medicine

基　　金：公安部重点课题(978221401)

摘　　要：研制人转铁蛋白的单链抗体。以纯化人转铁蛋白免疫小鼠,制备小鼠脾脏 mRNA,RT-PCR 扩增重轻链可变区基因;Linker 连接及引入 Sfi Ⅰ和 NotⅠ酶切位点构建单链抗体基因;同载体 pCANTAB 5E 连接,转化 E.coliTG1细胞,以 M13K07挽救制备噬菌体抗体呈现文库,筛选抗原阳性重组噬菌体粒子(Panning);感染 E.coliHB2151菌株,制备可溶抗体并检测其性质;测定抗体基因序列并分析同源性及结构特征。制备出4株分泌抗人转铁蛋白可溶抗体的 E.coli HB2151菌株。抗体基因只由小鼠抗体的轻重链可变区基因构成,开放读码,无终止子,并紧接一末尾为一琥珀终止密码的 E 末端序列,其对应氨基酸序列 V_H 和 V_L 部分均遵从抗体可变区特征。实验证明,可把基因工程抗体技术应用于法医血清学,以制备新型抗体试剂。To prepare human transferrin ScFv antibodies.The mRNA from the spleens of mice im- munized by the purified human transferrin was isolated.The genes of the variable region of either heavy or light chain were amplified by RT-PCR followed by linking with a linker and then the restriction sites of the Sfi Ⅰ and Not Ⅰ enzymes were introduced.The genes of the single chain variable fragment which is called ScFv was constructed.After that,the ScFv products were linked with the vector pCANTAB 5Es and then transforme...

关 键 词：抗人转铁蛋白 单链抗体(ScFv) 噬菌体呈现抗体库 固相抗原选择 

分 类 号：D919[医药卫生—法医学]