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Generation of monoclonal DNP-specific IgM and IgE murine antibodies on the efficacy of hybridization.
Murine hybridomas secreting monoclonal antibodies against the 2,4-dinitrophenyl (DNP) group were generated by fusion of NS1 mouse myeloma cells and mouse spleen cells. The spleen cells were obtained from recipient mice which had received spleen cells from syngeneic mice immunized with DNP-ovalbumin and which were subsequently infected with N.b. larvae simultaneously with the adoptive cell transfer. The recipient mice were immunized with DNP-N.b.-protein. Four fusion experiments were performed starting with spleen cells from recipient mice at days 5, 11, 20 and 44 after the adoptive cell transfer. Sixty-one hybrids out of 550 secreted immunoglobulins of an unknown specific including 4 IgE- and 7 IgM-secreting hybrids; 2 produced immunoglobulins of an unknown isotype. IgE clones were obtained only when the spleens were taken 1-2 weeks after the adoptive cell transfer, while only IgM-producing cells were generated with spleen cells 6 weeks after the adoptive cell transfer although the IgE-serum titers did not decline within this time period. The analysis of the light chains for the DNP-specific antibodies revealed seven with kappa- and three lambda-light chains.
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