Preparation And Potential Identification Of The Monoclonal Antibodies Against Psendorabies Virus FA

Pseudorabies virus(PrV), a member of Alpha Herpesvirus, is the causative agent of Pseudorabies (Aujeszky's disease),one of the most serious infectious diseases of several domestic and wild animals. Swine was the host and reservoir of PrV. Pseudorabies was an economically important disease of swine industry. Currently it is an important task to prevent, control and eradicate this disease in China.Accurate, promptly examines the cause of disease prevents one which of important measures disease proliferates. At present there are many diagnostic methods about PR,but there are a few diagnostic methods which was convenient,simple,low price and hypspecification.In many diseases detection,Mab was used generally which to show high-sensitivity and hadro-specificity.Less application of monoclonal antibody to detect PRV in China also reported. In order to benefit from the PRV infected animals and meat in the detection of food pathogens, Preparation and characterization of the experimental anti-pseudorabies virus antibody hybridoma cell line was carried out for follow-up to do a good job paving the way to start the experiment.With PK-15 cell culture PRV- FA longitude difference fast offcenter and sucrose density gradient offcenter, purification antigen. This experiment simultaneously prepares the Pseudorabies Virus cell culture medium and the chicken embryo culture medium, takes the immunity original immunity BALB/C mouse with the false mad dog chicken embryo culture medium, but takes indirect ELISA with the purity higher false Pseudorabies Virus's cell culture medium envelope antigen establishment indirect ELISA, after using for screens the fusion hybridoma cell, as far as possible avoids the non-specificity protein to the screening disturbance, cuts the false positive probability, enhances a masculine screening the efficiency and the accuracy. Takes the immunity BALB/c mouse's spleen cell and SP2/0 myeloma cytomixis application indirect ELISA screens, undergoes 3 time limited dilution method clones, obtained 2 hybridomas to be able to secrete the anti-pig false Pseudorabies Virus protein the monoclonal antibody hybridoma cell line, the naming is 4G9E9,1H9C9 stably, afterward carried on the appraisal to these two monoclonal antibody hybridoma cell line, after appraised these 2 hybridoma cell line is the IgG1 subgroup, 4G9E9 and in the 1H9C9 hybridoma cell culture clear and the mouse ascites monoclonal antibody ELISA titre respectively is 1:6400 and 1:12800 as well as 1:25600 and 1:102400. 2 monoclonal antibodies do not have the cross reaction with PPRV, HCV,JEV,PPV. The continuous cultivation 15 generations, 2 hybridoma cell lines still could stabilize the secretory antibody and the titre are invariable. Ascites after ammonium sulfate precipitation dialysis purification, the protein content is 2.412 mg/mL. These 2 monoclonal antibody hybridoma cell line's acquisition was the PRV research, the fast diagnosis method establishment has laid the good foundation.