基因免疫制备抗SARS病毒N蛋白单克隆抗体

目的通过基因免疫方法，制备抗SAILS病毒N蛋白的单克隆抗体并对单抗进行初步鉴定。方法将含有N蛋白的编码基因的真核重组质粒pSecTagB／N免疫BALB／c小鼠，然后取鼠脾细胞与骨髓瘤细胞融合，用原核表达的SARS病毒N蛋白筛选阳性克隆。然后通过免疫印迹法进一步证明抗体对N蛋白的特异性，用ELISA方法检测抗体对灭活SARS全病毒的免疫反应。结果筛选得到10株抗N蛋白的单克隆抗体，ELISA和免疫印迹结果表明这些抗体特异性针对N蛋白上的抗原决定簇，且有7株抗体对SARS灭活病毒有阳性反应。结论所得的单克隆抗体特异针对N蛋白。

Preparation of Monoclonal Antibodies Against N Protein of SARS Virus

Objective To produce monoclonal antibody against N protein of SAILS virus. Methods BALB/c mice were immunized with the recombinant plasmid pSeeTagB/N. Immune spleen cells were collected and hybridoma ceils were obtained by fusing the immune spleen cells with SP2/0 cells. Hybridoma cells producing monoclonal antibodies against N protein were screened by ELISA .The specificity of the antibodies to N protein was verified by Western blot. Results Ten hybridoma clones secreting antibody against N protein were obtained. N protein specific antibpdy was also detected in the mouse ascites fluid. Seven clones were capable to react with the inactivated SAILS coronavirus. Conclusion Hybridoma cells capable to produce monoclonal antibodies specific for N protein and the inactivated SARS coronavirus were produced.