抗地高辛人源小分子抗体的构建.pdfVIP

乔媛媛等　抗地高辛人源小分子抗体的构建　第 2 期 1·67 · 抗地高辛人源小分子抗体的构建 ( ) 乔媛媛 　王 　琰 　赵晓航 　张国民 　陈宇萍 　 海军总医院中心实验科 ,北京 100037 　　中国图书分类号 　R392 　　文献标识码 　A 　　文章编号 　1000484X( 2009) [摘 　要] 　目的 :将从大容量噬菌体抗体库中筛选得到的人源抗地高辛单链抗体进行分泌表达 ,并构建 Diabody (双体抗 ) ( ) 体 。方法 :将抗地高辛的噬菌体抗体转染 HB2151 菌 无琥珀抑制基因的宿主菌 ,IPTG 诱导 ,制备抗地高辛的可溶性单链抗 体。选取活性较好的克隆进行基因改造 ,构建 Diabody 表达载体 ,并分泌表达 ,表达 Diabody 进行 Western blot 鉴定。结果 :经 ELISA 结合活性测定 ,得到了结合活性较高的抗地高辛的可溶性单链抗体 ;基因改造后 ,经 ELISA 及 Western blot 测定证实获得 了结合活性较好的抗地高辛Diabody 。结论 :实验获得了分泌表达活性可靠的人源性抗地高辛抗体 ,并改造成应用前景较好的 Diabody ,为临床诊断及治疗洋地黄类药物中毒提供具有实用价值的人源小分子抗体。 [ 关键词] 　抗地高辛抗体 ;单链抗体 ;Diabody Construction of soluble antidigoxin ScFv antibodies and diabody QIAO YuanYuan , WANG Yan , ZHAO XiaoHang , ZHANG GuoMin , CHEN YuPing . Navy General Hospital , Beijing 100037, China [ Abstract] 　Objective :To clone soluble human antidigoxin antibodies from large phage antibody library and construct a vector that ex presses diabody. Methods :Soluble ScFvs were prepared through infecting E coli. HB2151 with the selected phage antibodies and induced with IPTG. The diabody vector was modified by enzyme digestion and checked by SDS electrophoretogram. Results :It was showed by ELISA that soluble ScFv had specific binding ability to digoxin. The vector to express a soluble diabody was obtained by genetic modification ,which was shown by Western blot. Conclusion :Soluble human antidigoxin antibodies were successfully obtained from phage antibodies. The vector is effi cient in creating diabody. [ Key words] 　Antidigoxin ;ScFv ;Diabody ( ) 　　地高辛 Digoxin 属洋地黄强心苷类化合物 , 临 子 , 提高抗体的效价 , 使其更有效地应用于临 床上作为正性肌力强心剂应用 ,但由于个体差异及 床[7 ,8 ] 。