Production of Monoclonal Antibodies Specific for Xanthomonas campestris pv, phaseoli

Authors: Fatma YÜCEL, Engin BERMEK, Beyazıt ÇIRAKOĞLU

Abstract: Xanthomonas campestris pv, phaseoli is a plant pathogen which causes common blight in beans. A hybridoma clone secreting a monoclonal antibody (6F3) against X. cam, pv, phaseoli was developed by fusion of the spleen cells of an immunized BALB/c mouse with FO (ATCC CRL 1646) mouse myeloma cells and possessing the characteristics specifed by the ELlSA system for use in the development of diagnostic kits. Hybrid cells resulting from fusion were detected in 348 wells. ELlSA tests showed one of these hybrids to react specifically with X. cam. pv. phaseoli. Limiting dilutions were performed in order to obtain a hybridoma cell culture producing monoclonal antibodies against X. cam. pve. phaseoli. The monoclonal antibody obtained. 6F3, had a higher affinity for the X. cam. pv. phaseoli (NCPPB-3035) used in the immunization. Its reaction with the other Xanthomonas strains and certain other bacteria strains was negligible. The purification of the monoclonal antibody and its isotype, the IgM subisotype are also reported. Monoclonal antibody thought to react with a membrane polypeptide antigen.

Keywords: Immunodiagnosis, monoclonal antibody , Xanthomonas campestris pv. phaseoli, ELlSA