2 CHICKEN B-CELL LINES RESISTANT TO OUABAIN FOR THE PRODUCTION OF CHICKEN MONOCLONAL-ANTIBODIES

Two chicken B cell lines, termed MuH1 and MuH4, which are resistant to ouabain, were established as fusion partners for production of chicke n monoclonal antibody from the parental cell lines R27H1 and R27H4 whi ch were deficient in thymidine kinase activity. The MuH1 synthesized, but did not secrete mu chain, whereas the MuH4 secreted non-specific I gM. Fusions of the MuH1 and MuH4 with spleen cells from chickens immun ized with human IgG resulted in successful preparations of hybridomas secreting antibodies specific to human IgG. Transformed cells insensit ive to HAT appeared in many culture wells after cell hybridizations wi th R27H4, but on hybridization with MuH1 or MuH4 the appearance of suc h unfavorable cells could be avoided by the inclusion of ouabain in HA T medium. Thus, in comparison with the R27H4 line, the MuH1 and MuH4 l ines did not provide higher fusion efficiencies, but gave increased op portunities to obtain hybridomas producing specific antibodies. Among a total of eight hybridomas, four derived from HuM1 and four derived f rom MuH4, seven secreted both IgG and IgM, and the remaining one deriv ed from MuH1 secreted only IgG. In all eight hybridoma lines IgG was s pecifically reactive to human IgG.