Monoclonal antibodies supernatant culture media

The hybridoma cells are initially grown in a medium that will not maintain the growth of the cancer cells these therefore die, as do non-fused lymphocytes, leaving only the fused cells. As the hybridoma cells grow, the supernatant fluid is tested for the presence of antibodies. Those cultures producing the desired antibody are further cloned and either grown in bulk or as tumours in animals and the monoclonal antibodies harvested. 

FIGURE 52-2 Generation of monoclonal antibodies. Mice are immunized with the selected antigen, and spleen or lymph node is harvested and B cells separated. These B cells are fused to a suitable B-cell myeloma that has been selected for its inability to grow in medium supplemented with hypoxanthine, aminopterin, and thymidine (HAT). Only myelomas that fuse with B cells can survive in HAT-supplemented medium. The hybridomas expand in culture. Those of interest based upon a specific screening technique are then selected and cloned by limiting dilution. Monoclonal antibodies can be used directly as supernatants or ascites fluid experimentally but are purified for clinical use. HPRT, hypoxanthine-guanine phosphoribosyl transferase. 

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