The Study Of CSFV McAb

Classical swine fever(CSF), a highly-infectious disease, brings severe economic losses (?)o the pig farming worldwide.It has been the most important infectious disease in our country for many years,and CSFV is the pathogeny.Continuous fever, capaillary vessel degeneration , hemorrhage, infarct and necrosis is the main pathological characters. In order to control CSF, many countries have paid a lot of manpower and money. In recent years, this disease has showen many new characters. The incidence of this disease raised, the clinical representation became complex and the incidence of the immuned hogs raised. CSF is still the most important infection in our country.Monoclonal antibodies are produced by cell fusion technology. In contrast to polyc(?)nal antibodies, monoclonal antibodies have following virtues: identical structures, un(?) component, high specificity, animate biotic-activity and fewer crossing reac(?) Monoclonal antibodies of CSFV are prospective in this study, with the purpose of prevent and diagnose.To gain purified special mice anti-CSFV, six Balb/C mice were immunized . Antibody titers were determined using indirect ELISA with CSFV E2 in coating the micro-wells. The specific anti-CSFV sera were detected in all mice and were sufficient for subsequent experiment.SP₂/O myelomas cell were chosen for cell fusion. Keep it in the bloom of proliferation before cell fusion.Feeder cells should be prepared before cell fusion. Kill one or more Balb/C mouse and draw its celiac macrophage and lymphocyte, then pave them in the cell culture wells. These should be operated one day before cell fusion.Electro-fusion, laser-fusion, chem.-fusion and bio-fusion are optional Chem-fusion is chosen. Highly purified Polyethylene 1000(PEG-1000) was used for cell fusion.Cell fusion was proceeded in a 50-ml centrifugal tube. Mingle spleen cells and myeloma cells (10:1) put into the centrifuge tube, and then instill PEG-1000 scrupulously at a c(?)ain dose. Put the mixed cells into the cell culture wells that had been paved with feeder cells., and selected-culture with HAT and HT. Avoiding contamination is cruciai to the cell fusion. Inspect the celis next day.When cells have paved the whole cells, select positive cells using indirect ELISA. And then proliferate the positive cells.Subclone the positive cells by limiting dilution for three times to get identical cells. And then inject them into mouse abdominal cavity. Retrieve ascites in the fifteenth day.After repetitious efforts, we got four positive hybridomas named 3D5,3D7,3F8 and 4B2 with indirect tiers of 1:102400-1 : 6400, which fulfill the expectation.The indirect immunofluorescent assay(IFA) results indicate that the four panels of mAbsreacted with the PK-15 cells, which were infected with CSFV Shimen and the CSFV E2 protein. In addition, All the mAbs, showed crossing reactivity with bovine viral diarrhea virus(BVDV) antigen.