Harding M J, Molitor T W
Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul.
Arch Virol. 1992;123(3-4):323-33. doi: 10.1007/BF01317267.
Monoclonal antibody technologies were applied to the study of early events in porcine parvovirus (PPV) infections in vitro. Balb/c mice were immunized with whole swine testicle cells and hybridomas were produced following fusion with myeloma cells. Resultant clones were screened firstly in an ELISA system, to detect monoclonal antibody recognition of swine testicle cells, and secondly, in a fluorescent antibody test to detect monoclonal antibody which inhibited production of PPV antigen. One clone, 1H11, which satisfied these screening requirements, recognized proteins present in cell lines both permissive and non-permissive for porcine parvovirus replication and inhibited the production of virus progeny of several PPV isolates. A linear staining pattern of cross-linked plasma membranes, indicative of monoclonal antibody binding at the cell membrane, was demonstrated by indirect immunofluorescence assays. In immunoblotting experiments, 1H11 recognized a polypeptide of approximately 40 kDa in size, present in both permissive and non-permissive cell lines.
单克隆抗体技术被应用于体外猪细小病毒(PPV)感染早期事件的研究。用全猪睾丸细胞免疫Balb/c小鼠,并在与骨髓瘤细胞融合后产生杂交瘤。首先在ELISA系统中筛选所得克隆,以检测单克隆抗体对猪睾丸细胞的识别,其次在荧光抗体试验中筛选,以检测抑制PPV抗原产生的单克隆抗体。一个满足这些筛选要求的克隆1H11,识别猪细小病毒复制允许和非允许细胞系中存在的蛋白质,并抑制几种PPV分离株病毒后代的产生。间接免疫荧光试验显示,交联质膜呈线性染色模式,表明单克隆抗体在细胞膜上结合。在免疫印迹实验中,1H11识别在允许和非允许细胞系中均存在的大小约为40 kDa的多肽。