摘要
分别用纯化的猪繁殖与呼吸综合征病毒（PRRSV）和纯化的重组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）。用纯化的PRRSV免疫小鼠，经细胞融合获得2株可分泌特异性单抗的杂交瘤细胞株，分别命名为4B8、4D8。用重组N蛋白免疫的小鼠，经细胞融合获得3株可分泌特异性McAb的抗PRRSV N蛋白的杂交瘤细胞2F3、4D5、5D11。间接ELISA检测4D8、4B8、4D5和5D11杂交瘤上清效价为1∶32～1∶512，而2F3的腹水效价为1∶12 800。单抗2F3、4B8和4D8与纯化病毒的Western blotting反应都为阳性，而4D5和5D11为阴性。IFA检测结果5株单抗都有明显的荧光，与PRRSV呈阳性反应。2F3的Ig亚型为IgM。5株单抗杂交瘤细胞连续传代至20代，分泌相应McAb的效价基本一致。本研究为PRRSV生物学诊断和方法研究提供有用工具。
After the immunization of BALB/c mice with purified porcine reproduction and respiratory syndrome virus （ PRRSV）, the stimulated splenocytes were fused with SP2/O myelomas to produce hybridomas. Two hybridoma lines of monoclonal antibody against N protein of PRRSV have been developed, designated as 4B8 and 4D8. Using recombinant N protein as immunogen, three hybridoma lines designated as 2F3, 4D5 and 5D11, were obtained. McAb 2F3 was identified as subclass IgM and the light chain belongs K chain. The ELISA titers of antibodies to PRRSV in cultural cell superuatant were 1：32 ～ 1：512, and the ELISA titer of the ascites of 2F3 was 1：12800. The results of IFA showed the McAbs could react with PRRSV in MARC-145 cells. Results of western-blotting with purified PRRSV indicated that the McAbs 2F3, 4B8 and 4D8 were against the lined epitopes in N protein of PRRSV. Together, it suggested that the McAbs would be useful reagents for diagnosis and epitope identification of PRRSV.
出处
《畜牧与兽医》
北大核心
2007年第4期1-4,共4页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金[30270990]
教育部博士点基金项目[20060307007]
新世纪优秀人才支持计划[NCET-04-0502]基金资助
