杆状病毒GP64蛋白单克隆抗体的制备及鉴定

Preparation and identification of monoclonal antibodies against baculovirus GP64 protein

为研究杆状病毒GP64蛋白在感染宿主过程中的表达情况,通过克隆构建含GP64基因的重组质粒,在CHO-S细胞表达重组GP64蛋白,利用野生型杆状病毒免疫BALB/c小鼠血清,筛选出重组GP64蛋白的单克隆抗体。结果表明,构建出正确的GP64重组质粒,成功表达出GP64蛋白,筛选出1株分泌单克隆抗体的杂交瘤细胞5B1;单克隆抗体亚类鉴定结果显示,单抗亚类为IgG2b,轻链类型为κappa;IFA和Western-blot结果显示筛选出的GP64单克隆抗体可以与杆状病毒及GP64蛋白发生特异性结合。结论,GP64蛋白的单克隆抗体制备成功。

In order to study the expression of baculovirus GP64 protein during host infection,a monoclonal antibody that recognized GP64 protein need to be prepared.In this study,a GP64 recombinant plasmid was constructed,and CHO-S cells was used to express the recombinant GP64 protein.Wild type baculovirus was used to immunize BALB/c mice,and recombinant GP64 protein was used to screen for monoclonal antibodies.In result,the correct GP64 recombinant plasmid was successfully constructed and the GP64 protein was successfully expressed.A hybridoma cell 5B1 that secreted a monoclonal antibody was selected.The results of series experiments showed that this monoclonal antibody subclass was IgG2b and the light chain type wasκappa.The results of IFA and Western-blot showed that the GP64 monoclonal antibody were specifically bonded to baculovirus and the recombinant GP64 protein.In conclusion,GP64 protein monoclonal antibody was successfully prepared.