茶尺蠖核型多角体病毒多角体蛋白单克隆抗体的制备

Preparation of a monoclonal antibody against polyhedrin of Ectropis obliqua nucleopolyhedrovirus

为了建立一种基于免疫反应检测茶尺蠖核型多角体病毒的方法,以纯化后的茶尺蠖核型多角体病毒作为抗原,免疫BALB/c小鼠,将小鼠脾脏细胞与小鼠骨髓瘤细胞Sp2/0融合,经间接ELISA筛选及克隆得到了一株稳定分泌单克隆抗体的杂交瘤细胞株,命名为7D3。同时克隆并在大肠杆菌中表达了EoNPV多角体蛋白基因,获得重组多角体蛋白。经Western blotting鉴定,该抗体可与EoNPV的多角体蛋白特异性结合。利用制备EoNPV多角体蛋白的单克隆抗体,建立了间接ELISA测定EoNPV的方法。

To develop a method based on immunoreactions for detection of Ectropis obliqua Nucleopolyhedrovirus（EoNPV）,the polyhedra of the virus were purified and used to immunize the mouse BALB/c.The spleen cells from the immunized mice were then fused with the myeloma cell line Sp2/0.A hybridoma cell line which can stably secrete the monoclonal antibody against EoNPV was achieved by using indirect ELISA screening and cloning methods,and was named as 7D3.Meanwhile,the polyhedrin gene was cloned from EoNPV and expressed in E.coli.Western blotting analysis showed that the monoclonal antibody prepared from 7D3 could specifically react with the recombinant polyhedrin.An indirect ELISA method based on this monoclonal antibody for detecting EoNPV in infected tea looper was developed.