猪脑心肌炎病毒VP1蛋白单克隆抗体的制备及鉴定

Preparation and identification of monoclonal antibodies against VP1 protein of porcine encephalomyocarditis virus

利用纯化的猪脑心肌炎病毒(EMCV)VP1重组蛋白免疫BALB/c小鼠,采用杂交瘤细胞技术,获得了2株稳定分泌抗EMCV VP1蛋白单克隆抗体的杂交瘤细胞株,分别命名为10B和11D。经ELISA测定,2株杂交瘤细胞培养上清效价分别为1∶1 600和1∶800,接种小鼠的腹水效价分别为1∶2.56×106和1∶1.28×106。2株单抗的亚类均为IgG2a,均能特异性识别重组VP1蛋白,但它们识别VP1蛋白的不同表位。间接免疫荧光和免疫组织化学试验证实,2株单抗具有良好的特异性,均能够识别EMCV。

Five 8-weeks-old BALB/c mice were immunized with purified recombinant fusion protein VP1 of porcine encephalomyocarditis virus（EMCV）. Two hybridoma cell lines designated 10B and 11D steadily secreting monoclonal antibody（McAb） against VP1 protein of EMCV were obtained by hyhridoma technology. The titers of the two McAbs in the supernatant culture supernatant were 1 ： 1 600 and 1 ： 800, respectively, and the titers in the ascites of the mice were 1：2. 56×10^6 and 1 ： 1. 28×10^5 respectively, detected by ELISA. The two McAbs helonged to IgG2a subclass and recognized different epitopes of VP1. The McAbs were able to recognize the recombinant VP1 protein of EMCV by Westernblotting, and could react with EMCV by indirect immunofluorescence assay （IFA） and immunohistochemistry, indicating that the two McAbs exhibit good specificity.