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  

Development of Monoclonal Antibodies Against N Protein of Porcine Reproduction and Respiratory Syndrome Virus

作　　者：丛晓燕[1,2] 吴家强[1,2] 李俊[1,2] 时建立[1,2] 杜以军[1,2] 孙文博[1,2] 尹斐斐[2,3] 谷琳 王金宝[1,2] 

机构地区：[1]山东省农业科学院畜牧兽医研究所,山东济南250100 [2]山东省畜禽疫病防治与繁育重点实验室,山东济南250100 [3]青岛农业大学,山东青岛266109 [4]新泰市新汶办事处,山东新泰271200

出　　处：《家畜生态学报》2011年第6期61-65,共5页Journal of Domestic Animal Ecology

基　　金：农业科技成果转化资金项目(2010GB2C600259);山东省自主创新成果转化重大专项(2010ZHZX1A0417);山东省农业重大应用技术创新课题(鲁财农便〔2010〕5号);山东省中青年科学家科研奖励基金(BS2009NY010)

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 1A12,3Fll, 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 1A12,3Fll, 5A3,4E4 reacted with cells infceted by PRRSV positively and the ELISA showed no reaction with antiserum to pseudorabies virus, Japanese epidemic encephalitis virus B, porcine circovurus virus, foot and mouth disease virus.

关 键 词：猪繁殖与呼吸综合征病毒 N蛋白 单克隆抗体 

分 类 号：S811.6[农业科学—畜牧学]