Analysis of anti-glycophorin monoclonal antibodies by flow cytometry

Our previous efforts with monoclonal antibodies against red blood cell antigens generally involved flow cytometric analysis and fluorescence activated cell sorting of erythrocytes using immunolabeling [1, 2, 3]. Therefore, our analyses performed for this workshop were by flow cytometry. Because flow cytometry is a technically exact method for measuring fluorescence from a large number of cells, specificity and sensitivity of labeling can be measured with high precision. However, the values determined with this technique may be much different from specificity and sensitivity determined using other measurements (e.g., hemagglutination or immunoblotting) or under different labeling conditions (e.g., different pH or ionic strength). Thus, we emphasize that evaluation of antibody characteristics depends on the measuring system.