INFLUENCE OF FLUORESCENT-ANTIBODY PROBE SPECIFICITY ON FLOW CYTOMETRIC ANALYSIS OF ANTIBODY-PRODUCING CELLS

In the flow cytometric analysis of the stability of antibody-producing cells, fluorescent antibody probes specific for immunoglobulin heavy chains have been widely utilized to quantify intracellular antibodies. To investigate the effect of the specificity of antibody probes on fl ow cytometric analysis, non-producing subclones of the 6-31 transfecto ma were stained with fluorescein isothiocyanate (FITC)-conjugated anti -human IgGs specific for heavy chain and light chain, respectively. Th e use of heavy chain-specific probe identified heavy chain-only produc ers as producers, whereas the use of light chain-specific probe identi fied light chain-only producers as producers. Thus, both heavy chain-s pecific and light chain-specific antibody probes should be used for th e accurate evaluation of a heterogeneous non-producing population. Fur thermore, the results of the flow cytometric analysis were confirmed b y immunoblotting, suggesting that flow cytometry is a useful technique for the rapid evaluation of the stability of transfectomas producing chimeric antibody.