鼠抗人转铁蛋白单链抗体基因的表达  

The expression of genes of single chain variable fragments of mouse's antibodies against human transferrin

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

机构地区：[1]公安部物证鉴定中心,北京100038

出　　处：《刑事技术》2002年第1期3-6,共4页Forensic Science and Technology

摘　　要：目的 制备抗人转铁蛋白单链抗体。方法 拼装好并带有酶切位点粘性末端的鼠抗人转铁蛋白的单链抗体基因经凝胶电泳定量后,同载体pCANTAB 5E连接,以其转化E.coli TG1细胞,经辅助噬菌体M13K07挽救构建噬菌体抗体表面呈现文库、抗原包被固相材料富集选择阳性重组噬菌体克隆（Panning）。感染能产生可溶抗体片段的大肠杆菌菌株E.coli HB2151,制备可溶抗体,以Anti-E末端抗体进行Western blot、ELISA检验和分子量测定。结果 人转铁蛋白的单链抗体基因成功表达。结论 用基因工程抗体技术制备抗体是可行的。OBJECTIVE To prepare single chain variable fragments of antibodies against human transferrin. METHODS The assembled genes of single chain variable fragments of mouse' s antibody against human transferrin, which are cohesive at the each of its two ends, were quantitated in agarose gel by electrophoresis, then ligated with vector pCANTAB 5E. The E. coli TGI cells were transformed by the ligated vectors, and reinfected with the helper phage M13K07 to rescue the recombinant phagemid to fabricate the phage-displaying antibody repertoires. Through a process called panning, the antigen - positive recombinant phage clones were selected and enriched, and with them the cells of E. coli HB2151, the strain which has capability to secrete soluble antibodies, were infected, resulting the soluble ScFv antibodies. By Western blot, ELISA with the addition of anti - E tag antibodies and molecular weight measurement, partial characterization to the antibodies newly-produced was carried out. RESULTS ScFv antibodies against human transferrin were produced successfully. CONCLUSION Preparing antibodies by genetic engineered antibody technology is feasible.

关 键 词：连接 转化 噬菌体抗体库 基因表达 可溶抗体 鼠抗抗体蛋白 单链抗体基因 法医物证 蛋白检验试剂 

分 类 号：D919.2[医药卫生—法医学] Q511[政治法律—法学]