猪繁殖与呼吸综合征病毒单克隆抗体的制备和鉴定

Preparation and Identification of Monoclonal Antibodies against PRRSV

目的制备抗猪繁殖与呼吸综合征病毒(PRRSV)单克隆抗体并对其生物学特性进行鉴定。方法以纯化的PRRSV全病毒抗原免疫BALB/c小鼠,通过杂交瘤技术制备单克隆抗体,并测定其免疫球蛋白亚类、效价和特异性。结果成功获得2株能稳定分泌抗PRRSVM蛋白的单克隆抗体杂交瘤细胞,命名为3C3、3D2,经鉴定亚类均为IgG2a、kappa链。2株单克隆抗体腹水ELISA效价达105～107,IPMA效价达1:2560～1:10240,与猪瘟病毒、猪伪狂犬病毒、猪细小病毒、猪圆环病毒等无交叉反应。结论获得特异性针对猪繁殖与呼吸综合征病毒的单克隆抗体,为研究该病的快速诊断技术奠定基础。

Objective To prepare and identify monoclonal antibodies （McAbs） against PRRSV. Methods BALB/c mice were immunized with the purified PRRSV, and monoclonal antibodies were obtained by means of hybridoma technique. The resultant monoclonal antibodies were identified for subtype, titer and evaluated for specificity. Result Two hybridoma cell lines stably secreting specific McAbs against M protein of PRRSV were obtained, named as 3C3 and 3D2, which were identified as immunoglobulin G2a （IgG2a） kappa chain, and the ELISA and IPMA titers of ascites were 1 ： 105 to 1：107 and 1：2560 to 1 ： 10240 respectively. The prepared McAbs showed specific reaction against PRRSV, and not reacted with CSFV, PRV, PPV and PCV. Conclusion Two McAbs against PRRSV have been successfully prepared, further facilitating which may the study of rapid detection techniques for PRRSV.