猪繁殖与呼吸综合征病毒SX-1株N蛋白单克隆抗体的制备

摘要

用纯化的重组HIS标签N蛋白免疫BALB/c小鼠,采用杂交瘤技术制备抗PRRSV N蛋白的单克隆抗体(McAb),经细胞融合获得5株可分泌特异性McAb的抗PRRSV N蛋白的杂交瘤细胞1A12,3F11,5A3,5F9,4E4.其中1A12,3F11,5A3,4E4杂交瘤上清与HIS-N蛋白与GST-N蛋白两种抗原包被的ELISA反应均为阳性.5F9与HIS-N蛋白包被的ELISA反应为阳性,而与GST-N蛋白包被的ELISA反应呈阴性.单抗1A12,3F11,4E4,5A3与HIS-N与GST-N蛋白的Western blotting反应均为阳性,而5F9只与HIS-N反应.IFA检测显示1A12,3F11,5A3,4E4株单抗都有明显的荧光,说明其与PRRSV呈阳性反应.同时,ELISA检测又有很好的特异性.%After the immunization of BALB/c mice with recombinant HIS-N protein, the stimulated splenocytes were fused with SP2/0 myelomas to produce hybridomas. 5 hybridoma lines of monoclonal antibody against N protein of PRRSV have been developed, designated as 1A12, 3F11, 5A3, 5F9,4E4. The results of IFA showed the McAbs could react with PRRSV in MARC-145 cells. The results of western-blotting indicated that all the McAbs were against the lined epitopes in HIS-N protein and GST-N protein of PRRSV except 5A3 and 5F9 which only reacted with HIS-N protein. The IFA showed the 1A12,3F11, 5A3,4E4 reacted with cells infceted by PRRSV positively and the ELISA showed no reaction with antise-rum to pseudorabies virus, Japanese epidemic encephalitis virus B, porcine circovurus virus, foot and mouth disease virus.