7] The use of conventional antisera in the production of specific monoclonal antibodies

Abstract

We have shown here that conventionally produced antisera aids in the production of specific monoclonal antibodies. The complex of immunoglobulin plus antigen acted as an effective immunogen in both rats and mice. Stable cell lines producing anit-human alpha 2M monoclonal antibodies to numerous antigenic sites on the alpha 2M molecule have been generated by this method. This method has also been used to produce monoclonal antibodies to human complement components. This procedure of immunizing mice with an immunoprecipitated product derived from conventionally produced antisera is a unique approach in that the antigen is conveniently enriched without tedious and time-consuming biochemical purification. In addition, the use of the immunoprecipitated product in conjunction with the immunoprecipitating antisera allows for rapid screening of the hybridomas in the initial stages when cell growth and maintenance is critical. This method thus further simplifies the task of obtaining a specific monoclonal antibody from a complex mixture. A final consideration is the wide availability of conventionally produced antisera to numerous proteins and other biological substances that could be used in the production of monoclonal antibodies, and consequently these antibodies could be used in more detailed studies of these same molecules.