猪繁殖与呼吸综合征病毒NVDC-JXA1株膜基质蛋白单克隆抗体的研制

Development of Monoclonal Antibodies against Matrix Protein of PRRSV NVDC-JXA1 Strain

为制备抗猪繁殖与呼吸综合征病毒NVDC-JXA1株单克隆抗体,用纯化的猪繁殖与呼吸综合征病毒NVDC-JXA1株免疫BALB/c小鼠,最后一次免疫后第3天取其脾细胞与SP2/0细胞在聚乙二醇作用下融合,通过酶联免疫吸附试验(ELISA)和间接免疫荧光试验(IFA)筛选,以有限稀释法克隆3次,制备单克隆抗体,并对制备的单克隆抗体进行鉴定。结果表明获得了2株分泌PRRSV膜基质蛋白特异性抗体的杂交瘤细胞,命名为1D12、5H5,经鉴定其抗体亚型分别为IgG2b、IgM,2株均为κ链,腹水ELISA效价分别为1∶107和1∶105。该单克隆抗体与PRV、PPV、PCV、CSFV、JEV无交叉反应。作者成功制备了抗猪繁殖与呼吸综合征病毒NVDC-JXA1株膜基质蛋白单克隆抗体,为进一步建立相关诊断方法奠定了基础。

To develop and characterize monoclonal antibody（McAb） against porcine reproductive and respiratory syndrome virus NVDC-JXA1 strain（PRRSV NVDC-JXA1 strain） for related research work.BALB/c mice were immunized conventionally with the purified virions of PRRSV NVDC-JXA1 strain.On the third day after the final immunization,spleen cells of mice fused with myeloma cells SP2/0 were filtered by culturing selectively,diluting finitely,enzyme-linked immunosorbent assay（ELISA） and IFA test,to obtain the subtypes from the clone line by EI.ISA test. As a result, the cell lines secreted McAbs stably by culturing in vitro and recovery tests were obtained and named as 1D12, 5H5 respectively, the ELISA titers of ascites was 1 ： 10^7 ,1 ： 10^8 respectively. The subtypes of the two McAbs were identified as IgG2b and IgM,Kappa light chain. The two McAbs reacted specifically with only matrix protein of PRRSV but not with PRV,PPV, PCV,CSFV,JEV by indirect-ELISA. The McAbs will be very useful as specific reagents in diagnosis and control of PRRS.