A Novel System for the Quantification of the ADCC Activity of Therapeutic Antibodies

Figure 11

Quantification of the ADCC activity of trastuzumab using RTU iLite effector cells and ERBB2⁺ HEK293 target cells. Individual vials of frozen RTU iLite effector cells and vials of frozen RTU ERBB2⁺ or ERBB2⁻ HEK293 target cells from 3 different production lots were thawed, mixed at E:T ratio of 4 : 1, and incubated for 6 hours at 37°C in the presence of increasing concentrations of trastuzumab prior to the addition of Nano-Glo Dual-luciferase reagent (Promega, Madison, WI) and the sequential determination of FL and NL activity. Results are expressed as RLU (a) or fold induction relative to the control sample consisting of effector and target cells without trastuzumab (b).