Immunization and Fusion Protocols for Hybridoma Production

Abstract

Kohler and Milsteins’ (1) technique of monoclonal antibody production is now being exploited in most areas of biology. The essence of the method is to immortalize and then select for clones of plasma cells secreting antibody against a desired antigen. Individual plasma cells secrete antibody of a single antigenic specificity and monoclonal antibodies are thus obtainable from a cell line derived from a single plasma cell. Once established, clonal lines of hybridomas (hybrids composed of plasma cells fused with immortal myeloma cells) provide an infinite supply of antibodies with reproducible properties. Monoclonal antibodies may be produced without extensive purification of the immunogen, and without the tedious cross-adsorption steps often necessary for production of specific antisera. So far, only mouse and rat hybridomas are being produced routinely, while human monoclonal antibodies of potential therapeutic value are technically more difficult to obtain.