title: Antibody-dependent cell-mediated cytotoxicity: a flow cytometry-based assay using fluorophores.
abstract: 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.
authors.affiliation: Applied Development Laboratory, Imperial Cancer Research Technology, Dominion House, St Bartholomew's Hospital, EC1A 7BE, London, UK.
authors.forename: R W
authors.initials: RW
authors.lastname: Wilkinson
authors.forename: A E
authors.initials: AE
authors.lastname: Lee-MacAry
authors.forename: D
authors.initials: D
authors.lastname: Davies
authors.forename: D
authors.initials: D
authors.lastname: Snary
authors.forename: E L
authors.initials: EL
authors.lastname: Ross
mesh_terms: 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
chemical_list: D002232:Carbocyanines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C070080:PKH 26; C098830:TO-PRO-3