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Abstract Female, 6-8w Balb/c mice were immunized with the purified recombinant fusion protein. After three immunizations, the spleen cells from Balb/c immunized were fused with mouse myeloma cells SP2/0. An indirect ELISA with purified VP1 protein as coated antigen was used to screen antibody-producing hybridomas. By 3 cycles of limited dilutions, Six positive hybridoma cell lines stably secreting monoclonal antibodies were selected. All MAbs reacted with MDCC-MSB1 cell infected with CAV by indirect immunofluorescence assay (IFA) and they recognized the VP1 protein expressed in Baculovirus by Western blot. This demonstrated that all MAbs have good specificity. The subtype of all MAbs is IgG1 and light chains of all MAbs are kappa. Taking advantage of VP1 fragments expressed, the epitopes corresponding to the MAbs are analyzed. The results showed that the monoclonal antibodies 1C5、2F3、4D2 and 4E8 recognized the same epitope, which was located in 218-274aa. Moreover, the two other epitopes were identified and they were located in 274-301, 324-369aa, respectively.
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Received: 25 September 2006
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