Antibody-dependent cell-mediated cytotoxicity: a flow cytometry-based assay using fluorophores.
Using flow cytometry (FCM), an assay has been developed for the determination of antibody-dependent cell-mediated cytotoxicity (ADCC). Target cells were labelled with a membrane dye, PKH-26, to allow discrimination when incubated with effector cells and antibody. Post-incubation, cell death within the PKH-26+ target cell population was assessed by the addition of the viability 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 with the potential to be a useful tool for evaluating the therapeutic potential of antibodies and antibody-based reagents.
Wilkinson
R W
RW
Applied Development Laboratory, Imperial Cancer Research Technology, Dominion House, St Bartholomew's Hospital, EC1A 7BE, London, UK.
Lee-MacAry
A E
AE
Davies
D
D
Snary
D
D
Ross
E L
EL
D000818:Animals; D000920:Antibody-Dependent Cell Cytotoxicity; D002232:Carbocyanines; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D009930:Organic Chemicals; D015203:Reproducibility of Results
D002232:Carbocyanines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C070080:PKH 26; C098830:TO-PRO-3