الفهرس | Only 14 pages are availabe for public view |
Abstract A panel of monoclonal antibodies (MAbs) against each of Leptospira interrogans serovar Canicola and pathogenic Leptospira spp. were produced. L. interrogans serovar Canicola was cultivated, concentrated and prepared for fusion. BALB/c mice of 6 weeks old were inoculated with different antigenic preparations of L. interrogans serovar Canicola with complete freund’s adjuvant. Booster doses with and without incomplete freund’s adjuvant were applied. To obtain hybridomas, mice spleen cells were fused with myeloma cells in the presence of polyethylene glycol. Hybridoma cells that secret specific antibodies were selected on the basis of ELISA. These positive hybridomas were cloned by limiting dilution method to reveal subclones of one cell origin. 5 specific clones were obtained. Characterization of the produced MAbs was applied using ELISA, MAT and Western bot technique. The results revealed that three of the obtained MAbs were specific to pathogenic Leptospira spp. and two were specific to L. interrogans serovar Canicola. ELISA kits with different formats were made. Adapting the myeloma cells to medium containing 2-ME is highly recommended before fusion. |