Assessment of Antibody-Dependent Cellular Cytotoxicity by Flow Cytometry

Abstract

A useful feature of therapeutic antibodies is the ability to kill the cells to which they bind. Antibodies are capable of mediating cell killing in a variety of ways. Apoptosis, complement-mediated mechanisms, and antibody-dependent cellular cytotoxicity (ADCC) are all effects that can be assayed to characterize lead antibody candidates. Extensive, multidose characterizations of a series of candidates can be performed in a short amount of time using assays developed for high-throughput flow cytometry systems. Antibodies that contain the Fc portion of the human IgG1 can activate complement-mediated killing. In the ADCC method described here, cytotoxicity is mediated mostly by natural killer (NK) cells. Thus, if an antibody binds to its target on the surface of a tumor cell, Fc receptors on the surface of the NK cells (effector cells) recognize the bound antibody. This leads to the release of cytotoxic granules containing perforin, granzymes, and interferon γ, a cytokine that can stimulate other cells of the immune system such as T cells.