Rapid Screening and Replica Plating of Hybridomas for the Production and Characterization of Monoclonal Antibodies

This chapter describes the rapid screening and replica plating of hybridomas for the production and characterization of monoclonal antibodies. The chapter presents a report that mouse myeloma can be fused with antibody-producing spleen cells and that the resulting cloned hybrid cell lines “hybridomas” continued to secrete monoclonal antibodies. This discovery initiated a new era in immunological research. The chapter describes that the opportunities for future research and the clinical applications in both diagnosis and therapy of disease appear to be enormous. Details of the experimental procedures for the production of hybridomas are explained in the chapter. The chapter explains that the most difficult aspect of the hybridoma technique is the identification and selection of the hybridomas that are producing antibody of the desired specificity. In addition, many screening assays are used to identify the antibody-producing cultures, including a variety of radioimmunoassays, enzyme immunoassays, immunofluroscence assays, and hemolytic assay. Selection of the optimal screening assay is critical to the success of the development of a hybridoma. This selection depends heavily upon the physiochemical characteristics of the antigen of interest.