Intra-Species Mice Hybridomas Against a Recombinant Protein of Mycobacterium avium paratuberculosis

Abstract

Singh, M., Siddiqui, M.Z. and Singh, R.P. 2002. Intra-species mice hybridomas against a recombinant protein of Mycobacterium avium paratuberculosis. J. Appl. Anim. Res., 22: 137–144. Intra-species mice hybridomas were produced by fusing the 35-kDa recombinant protein of Mycobacterium avium paratuberculosis primed splenocytes of Swiss white mice with BALB/c derived myeloma cells. Presence of peritoneal macrophages derived from Swiss white mice as feeder layer did not make much difference in the production of intra-species mice hybridomas. The total hybridoma production efficiency was 71% when peritoneal macrophages were used as feeder layer, whereas it was 59% when no feeder layer was used. Five stable positive hybridomas were subcloned to produce monoclonals. Out of 5 monoclonal antibodies (MAbs) obtained, 4 MAbs (1AG2, 2BD2, 2BG2, 1DE8) were of IgGl sub-class, whereas the fifth MAb (1AG8) was IgG2b. The Western blot analyses of these MAbs demonstrated the specificity of the antibodies to the recombinant protein through the presence of strongly reacting bands on immunoblots.