Development of a sandwich ELISA based on the monoclonal antibodies against VP2 of infectious bursal disease virus

Three hybridomas secreting monoclonal antibodies(mAb)against VP2 of infectious bursal disease virus(IBDV)were established by fusing SP2/0with spleen cells from BALB/c mice immunized with virus-like-particle recombinant VP2 protein,designated as 1D11,2G8and 2E5.The subtypes of immunoglobulin were IgG2bκ,IgG2bκand IgG1κ,respectively.Indirect immunofluorescence assay(IFA)proved that the three mAb could react to VP2 specifically.Additivity ELISA revealed that three mAbs recognized spatially independent epitopes of VP2 .Neutralization test indicated that the neutralization titer of 1D11and 2G8was 104 and 103 respectively,while 2E5did not have neutralizing activity.A sandwich ELISA was established using 1D11as capture antibody and 2E5as enzyme-labeled antibody,and the detectable minimum of IBDV was 102 TCID50/mL.The detection kit of sandwich ELISA was developed and proved to be applicable to the clinical detection of IBDV.