抗博尔纳病病毒核蛋白单克隆抗体的制备和鉴定

Preparation and identification of monoclonal antibody against nucleoprotein of Borna disease virus

目的制备并鉴定鼠抗博尔纳病病毒核蛋白单克隆抗体。方法重组核蛋白免疫Balb/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞进行常规融合,用间接ELISA法筛选阳性杂交瘤并进行有限稀释法克隆和亚克隆,建立分泌抗核蛋白单克隆抗体的杂交瘤细胞株,纯化单克隆抗体并进行效价测定和特异性鉴定。结果建立了1株能稳定分泌抗核蛋白单克隆抗体的杂交瘤细胞株,命名为2F6E8,重链为IgG2a,轻链为κ。ELISA法测得腹水的效价高达1∶32 000以上。Western-blot和间接免疫荧光实验证实该单克隆抗体能特异性结合重组核蛋白及天然核蛋白。结论成功制备了效价高、特异性好的抗核蛋白单克隆抗体,为建立博尔纳病病毒血清免疫学检测方法奠定了基础。

This study aims to prepare and identify monoclonal antibodies against Borna disease virus（BDV） nucleoprotein.Balb/c mice were immunized with recombinant BDV nucleoprotein and the spleen cells of the immunized mice were fused with the SP2/0 myeloma cells.The positive clones were selected and subcloned 3 times.One hybridoma cell line secreting specific McAb against BDV nucleoprotein was obtained which named 2F6E8,and the McAb was belonged to IgG2a.ELISA showed that the titers of the ascites was 1∶32 000;Western blotting and IFA experiments illustrated that the monoclonal antibody could specifically bind to BDV nucleoprotein.This polyclonal antibody may provide the foundation for the further studies on BDV serum immunology detection methods.