Preparation of monoclonal antibodies against the MSP5 of Anaplasma marginale.

Monoclonal antibodies (MAbs) against MSP5 protein of Anaplasma marginale were produced by fusing mice myeloma cell with spleencytes of BALB/c mice immunized with recombinant MSP5 protein.Two hybridoma cell lines 1D8 and 2F3 that stably produce antibodies against MSP5 were identified by ELISA.The ascites antibody titers of the hybridoma lines were 5.49 ×105 and 7.83 ×104 respectively.The subtypes of MAbs were identified as IgG2b and IgG2a.Additive index of ELISA competitive binding assay indicated that both MAbs were able to recognize identical epitopes of the MSP5 protein of A.marginale.Western blot analysis showed that the MAbs specifically recognized certain antigenic epitopes of MSP5 protein of A.marginale,but not those of other reference antigens.