One important role of the antibody effector functions is complement-dependent cytotoxicity (CDC), which induces cell lysis by a cascade of activation triggered by the binding of C1q subunits to the Fc regions of antibodies bound to the cell surface. CDC can be modulated by either Fc isotype engineering or Fc genetic mutations or Fc glycosylation profile modifications. A luminescence method based on ATP measurement will be used to estimate cells damaged by CDC. The binding of C1q and C4b to the Fc fragment of antibody to will be evaluated by FACS.

Schematic of Complement-dependent Cytotoxicity:

Reference:

1. 1. Duensing TD and Watson SR. Complement-Dependent Cytotoxicity Assay. Cold Spring Harb Protoc; 2018(2).
   2. Broyer L, Goetsch L, Broussas M. Et al Evaluation of complement-dependent cytotoxicity using ATP measurement and C1q/C4b binding. Methods Mol Biol. 2013;988:319-29.
   3. https://www.promega.com/products/cell-health-assays/cell-viability-and-cytotoxicity-assays/celltiter_glo-luminescent-cell-viability-assay/?catNum=G7570