Abstract: Female, 6-8 weeks old BALB/c mice were immunized with the purified recombinant fusion VP3 of IBDV. After three immunizations, the spleen cells from immunized BALB/c mice were fused with mouse myeloma cells SP2/0. By 3 cycles of limited dilutions, we obtained four strains of hybridoma cells, named HRB7B,HAR-7C,HRB-3F and HRB-10E. Isomerism and stability analysis indicated that those monoclonal antibodies had specific reaction with the VP3 of IBDV. The titer of four antibodies were up to 5.0×10² in cell culture supernatants and 6.4×10⁶ in ascetic fluids. Plusing ELISA showed that in four cell strains, HRB-7C and HRB-10E have the similar antigen epitopes. Moreover, the study identified the antigenic epitopes by peptide scanning, further structural and function analysis of IBDV. This work could provide the basis for the development of immunitybased prophylactic, therapeutic and diagnostic techniques for the control of IBD and further structural and function analysis of IBDV.