Enhancing the frequency of antigen-specific hybridomas.

Techniques were developed to enhance the frequency of antigen-specific hybridomas by enriching for antibody precursor or-producing lymphocytes prior to cell fusion. Spleen cells of immunized animals were either (a) cultured in vitro for 3-4 days in the presence of the immunizing antigen or (b) adoptively transferred into X-irradiated syngeneic recipients followed immediately by an in vivo antigen boost 4 days prior to use in cell fusion. There was a 10-fold increase in the percentage of wells which develop antigen-specific, antibody-secreting hybridomas following these treatments. These techniques allow an increased yield of hybridomas with a greatly decreased technical effort.