Development And Primary Application Of Hybridoma Cell Lines Respectively Secreting Monoclonal Antibodies Against Infectious Bursal Disease Virus And Lipopolysa Ccharide Of Shigellae In The Chichen

Infectious bursal disease virus and Lipopolysaccharide of Shigellae in the Chicken were maded mixed antigens in this study.Mice was immunized with the mixed antigens,which added Bursin.Hybridoma cell strains secreting monoclonal antibodies against two antigens were obtained by one fusion and screen respectively methods. Compared with traditional method using sole antigen immunize mouse,this method used mix immunity,screening respectively,is a highly effective method in obtained the hybridoma cell strains.Hybridoma cell strains against different antigens can obtained just through one immunity and one fusion.This method not only has saved the experimental material,reduced the cost,moreover,it has saved the time and the manpower,greatly enhanced the working efficiency.Now one people could finish the work needed three or more people finished before.This has the vital significance to the scholarly research and the practical application of hybridlump technology.IBDV and LPS of Shigellae in the Chicken was purified by sucrose concentra-tion and differential centrifugation.Eight-week Balb/c mice were immunized according to Kazuhiro's produre.Murine splenocytes of the immunized mice after the third immunization were fused with SP2/0 myelomas,An indirect Enzyme-linked Immunosorbent Assay(ELISA)coated with He4 IBDV and LPS was used to screen hybridoma for production of specific antibody in hybridoma culture fluid.Three hybriddoma against Infectious bursal disease virus were generated,named 4H5,4B2,3C11,four hybridoma against Lipopolysaccharide were generated,named 3C2,3C4,4B2,4A3.The monoclonal antibodies(McAbs)were categorized into IgG2 subtype.The result of cross-reaction in ELISA dedicated that seven McAbs were type-sepecific.The indirect ELISA titers of the 4H5 hybridoma against Infectious bursal disease virus in hybridoma culture fluid and mouse ascites were 1:25600 and 1:102400 respectively.The ELISA titers of the 4H5 McAbs in hybridoma culture fluid and mouse ascites were 1:102400 and 1:819200 respectively.The cell strain also can stably secrete the McAbs after twenty-six serial passages and freezing-thawing three times within six months.Mouse ascites was purified and protein concentrations were 78.45mg/mL and 88.65mg/mL respectively.The obtained of the two species hybridoma cell strains against two antigens laied the foundations of study and rapid diagnosis of Infectious bursal disease virus and Shigellae in the Chicken.