尼帕病毒G和F蛋白单克隆抗体的制备与鉴定

摘要

To prepare the monoclonal antibody (Mab) against G or F protein of Nipah virus (NiV), hybridoma cell lines were produced by fusinig SP2/0 cells with spleen cells from mouse immunized with Pcagg-NiVG or Pcagg-NiVF expressing G or F protein of NiV, respectively. Two hybridoma cell lines, designed G3E9 and F2G1 were ^identified by indirect immunofluorescence assay, which were stablely secreting Mabs against G or F protein. The two Mabs were IgM subtype with kappa light chain. Both Mabs were able to reduce syncytium formation of BHK. Cells that induced by the co-expression of G and F protein of NiV. Futhermore, the Mabs also had the ability to neutralize the psudotyped vesicular stomatitis virus expressing both G and F proteins of NiV.%为制备抗尼帕病毒(NiV)G和F蛋白的单克隆抗体(MAb),本研究以表达NiV G和F蛋白的真核重组表达质粒pCAGG-NiVG和pCAGG-NiVF分别免疫BALB/c小鼠,采用常规技术制备杂交瘤细胞；以表达G和F蛋白的重组牛痘病毒(rWR-NiVG、rWR-NiVF)分别感染BHK细胞,通过间接免疫荧光(IFA)筛选阳性克隆,分别得到一株抗G和抗F蛋白的MAb,命名为G3E9和F2G1.两株MAb均为IgM亚型/κ链.IFA检测G3E9和F2G1杂交瘤细胞诱导的小鼠腹水中IgM抗体效价均为1:1 024.细胞融合抑制试验表明两株MAb均具有抑制NiV G和F蛋白共表达诱导细胞融合的功能；并且采用表达NiV G和F蛋白的水泡性口炎伪型病毒进行的中和试验表明两株MAb具有一定的中和活性.