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Specific monoclonal antibodies are produced by hybridoma cells, which form when B lymphocytes and myeloma cells combine.
Hybridoma or phage-display technology can be used to make monoclonal antibodies. In order to elicit an immunological response, a mouse is first immunized using a specific antigen in hybridoma technique. In hypoxanthine-aminopterin-thymidine (HAT) media, B cells from the spleen of the immunized mice are isolated and united with myeloma cells that have undergone immortalization. Myeloma cells are specifically chosen to be HAT-sensitive and non-antibody secreting. Hybridomas, the resulting fusion product, secrete antibodies and are immortal.
Unfused B cells also perish since they are not immortal and cannot survive in the HAT mixture with unfused myeloma cells. Then, these hybridomas are diluted into single cell wells and put through an antibody screening process. These cloned B cells generate particular monoclonal antibodies.
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