Antibody-dependent cell-mediated cytotoxicity: a flow cytometry-based assay using fluorophores

Using flow cytometry (FCM), an assay has been developed for the determinati on of antibody-dependent cell-mediated cytotoxicity (ADCC). Target cells we re labelled with a membrane dye, PKH-26, to allow discrimination when incub ated with effector cells and antibody. Post-incubation, cell death within t he PKH-26(+) target cell population was assessed by the addition of the via bility probe TO-PRO-3 iodide (TP3). This ADCC method allows analysis to be conducted on a single cell basis and overcomes the need for radiochemicals. This communication indicates that the assay is accurate and reproducible w ith the potential to be a useful tool for evaluating the therapeutic potent ial of antibodies and antibody-based reagents. (C) 2001 Elsevier Science B. V. All rights reserved.