Enhancing the frequency of antigen‐specific hybridomas

Abstract

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.