寨卡病毒NS1蛋白的原核表达和单克隆抗体的制备

Prokaryotic expression and monoclonal antibody preparation against NS1 protein of Zika virus

目的高效表达和纯化重组的寨卡病毒NS1蛋白,制备抗NS1蛋白的单克隆抗体。方法构建含有寨卡病毒NS1基因的原核表达质粒,利用大肠杆菌大量表达重组NS1蛋白,纯化蛋白后免疫小鼠并进行细胞融合,筛选制备高纯度的单克隆抗体。结果在大肠杆菌中高效表达了重组NS1蛋白,重组NS1蛋白具有良好的免疫原性,筛选出3株分泌针对寨卡病毒NS1蛋白单克隆抗体的杂交瘤细胞株,分别命名为6B8、7D11和3E2,纯化的单克隆抗体与重组NS1蛋白有良好的特异性反应。结论利用重组NS1蛋白免疫制备了抗NS1的单克隆抗体,为后续建立针对NS1蛋白的ELISA检测方法及相关研究提供了基础。

In order to prepare the monoclonal antibodies against Zika virus NS1 protein,a recombinant plasmid containing Zika virus NS1 gene was constructed,and the recombinant NS1 protein was expressed by E.coli.The mice were immunized with the purified recombinant NS1 protein,and anti-NS1 monoclonal antibodies were prepared by cell fusion and screening.After rounds of selection,three strains of hybridoma cell lines which secreted monoclonal antibodies against Zika virus NS1 protein were established and designated as 3E2,6B8 and 7D11 respectively.The monoclonal antibodies with high purity were generated and purified from the ascites.All the mAbs belong to IgG 1 isotype,and have a specific reaction to the recombinant NS1 protein.In conclusion,the monoclonal antibodies against NS1 were prepared,which will provide a raw material for the ELISA detection method of Zika virus.