Production of stable rabbit-mouse hybridomas that secrete rabbit mAb of defined specificity.
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.
Raybould
T J
TJ
Allelix Inc., Diagnostics Division, Mississauga, Ontario, Canada.
Takahashi
M
M
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002459:Cell Fusion; D002460:Cell Line; D006825:Hybridomas / Q000276:immunology*; D007621:Karyotyping; D051379:Mice / Q000276:immunology*; D011817:Rabbits / Q000276:immunology*; D013297:Streptococcus pyogenes / Q000276:immunology; D013997:Time Factors
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal