In Vitro Immunization for the Generation of Hybridomas Using Serum-Free Medium
No one engaged in research in the biological sciences is unaware of the impact that monoclonal antibodies have had as research tools. These proteins recognize specific sites on a single antigen and often bind to these sites with high affinity. Because of this, researchers have found many uses for monoclonal antibodies, including their application in immunoassays (1), purification or identification of cellular proteins (2,3), and studies of enzyme action (4). The significance of monoclonal antibodies was recently acknowledged when the 1984 Nobel prize was shared by the co-inventors of the B-B-cell hybridoma, Kohler and Milstein (5). As the commercial applications of monoclonal antibodies begin to grow, newer and better techniques for hybridoma formation will be necessary to meet the demand for antibodies with unique, well-defined specificities and distinctive immunological and physical properties. One approach to the improvement of hybridoma formation techniques is to attempt to define the components and reaction conditions that result in maximal formation of antibody-secreting hybridomas. Until recently, all aspects of in vitro immunization and hybridoma formation have involved an ill-defined component—animal serum, usually fetal bovine serum.
KeywordsConditioned Medium Spleen Cell Lymphoid Cell Ammonium Sulfate Precipitation Fusion Frequency
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