抗地高辛人源小分子抗体的构建

Construction of soluble anti-digoxin ScFv antibodies and diabody

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。

Objective：To clone soluble human anti-digoxin antibodies from large phage antibody library and construct a vector that expresses 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 anti-digoxin antibodies were successfully obtained from phage antibodies. The vector is efficient in creating diabody.