An alternative methodology for a quantitative flow-based cell-mediated in vitro cytotoxicity assay to evaluate immune cell potency

In vitro cytotoxicity assays have a crucial role in determining the functionality and potency levels of various immune cells, including both NK and T-cells. A flow cytometry based cytoxicity assay is commonly used to measure the levels of cell-mediated killing of a target cancer cell. The assay involves staining the cancer cells along with a co-culture incubation step. The cytotoxicity value is typically reported as a percentage killing efficiency of the target cancer cell at a fixed effector-to-target ratio. In vitro cytotoxicity assays have commonly been used to provide valuable potency data and have many diverse applications for multiple cell types. We propose an alternative assay methodology which can improve accuracy over traditional cytotoxicity analysis methods. This new reporting method/approach can overcome signal saturation issues and allows for more consistent and meaningful outputs over a broader dynamic assay range. It also eliminates any potential extrapolation needed during analysis. This new method provides a means for a more robust cytotoxicity assay, resulting in a more valuable measuring tool