Application of area scaling analysis to identify natural killer cell and monocyte involvement in the GranToxiLux antibody dependent cell-mediated cytotoxicity assay

Modification of the GranToxiLux ADCC assay to include Area Scaling Analysis (ASA) for qualitative assessment of antibody recruitment of NK cells and monocytes Conclusion: The inclusion of area scaling analysis (ASA) in the GranToxiLux ADCC assay allows simultaneous quantification of ADCC activity at the target cell level, and assessment of the contribution of natural killer cells and monocytes to the total observed ADCC activity when whole human peripheral blood mononuclear cells are used as a source of effector cells. The Duke Cytometer has been optimized and maintained according the procedures described by Perfetto and colleagues (Perfetto SP, Ambrozak D, Nguyen R, Chattopadhyay PK, Roederer M. Quality assurance for polychromatic flow cytometry using a suite of calibration beads. Nat Protoc 2012;7:2067-79.) In addition, both the Duke Cytometer and the IHV cell sorter are evaluated daily using Cytometer Setup & Tracking Breads – BD Biosciences.