Science, Vol 240, Issue 4860, 1788-1790
Copyright © 1988 by American Association for the Advancement of Science
Production of stable rabbit-mouse hybridomas that secrete rabbit mAb of defined specificity
TJ Raybould
and
M Takahashi
Allelix Inc., Diagnostics Division, Mississauga, Ontario, Canada.
Inclusion of normal rabbit serum (NRS) in culture medium after interspecific fusion of hyperimmunized rabbit spleen cells with murine SP2/0 myeloma cells produced 271 rabbit-mouse hybridomas (RMHs) that secreted rabbit immunoglobulin against group A Streptococcus (GAS). Continued use of NRS-supplemented medium during cloning yielded stabilized monoclonal RMH lines that have secreted GAS-specific rabbit antibody at concentrations similar to murine hybridomas (3 to 8 micrograms per 10(6) cells per 24 hours), for over 4 months of culture in vitro. The use of NRS as a medium supplement during initial culture, cloning, and stabilization of RMHs enables production of considerably more specific rabbit monoclonal antibody (mAb)-secreting RMHs than have previously been reported.
