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

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1D12、5H5，经鉴定其抗体亚型分别为IgG2b、IgM，2株均为κ链，腹水ELISA效价分别为1∶107和1∶105。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。

Development of Monoclonal Anti bodies against Matrix Protein of PRRSV NVDC-JXA1 Strain

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 ELISA test.As a result，the cell lines secreted McAbs stably by culturing in vitro and recovery tests were obtained and named as 1D12，5H5 respectively，the ELISA titers of ascites was 1∶107，1∶105 respectively.The subtypes of the two McAbs were identified as IgG2b 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.