[2] Methods of enhancing the frequency of antigen-specific hybridomas

This chapter discusses the methods of enhancing the frequency of antigen-specific hybridomas. The hybridization of spleen cells from immunized mice with plasmacytoma cell lines is a powerful method for the production of monoclonal antibodies. However, the development of monoclonal antibodies has been hampered by low frequency of antigen-specific hybridomas recovered after cell fusion. In an effort to improve the yield of hybridomas, the chapter discusses two techniques to select and expand the number of spleen cells producing specific antibody prior to fusion. The first method is the adoptive transfer of spleen cells from immunized animals to X-irradiated syngeneic recipients followed by in vivo antigen boosting. In the second method, spleen cells from immunized mice have been cultured with antigen before fusion. The use of either of these methods resulted in a 10-to 50-fold increase in the percentage of antigen-specific antibody-secreting hybridomas. These methods for enhancing the yield of antigen-specific hybridomas and the techniques used for cell fusion and cloning by limiting dilution are described in this chapter.