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

Figure 15

Quantification of the ADCC activity of cetuximab using iLite effector cells (V-variant) and EGFR⁺ HEK293 target cells: effect of incubation time. (a) RTU iLite effector cells V-variant (1.2 × 10⁵ cells/well) and RTU EGFR⁺ HEK293 target cells were mixed at an E:T ratio of 4 : 1 and incubated for 4, 6, or 18 hours at 37°C in the presence of increasing concentrations of cetuximab 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 FL activity in RLU. (b) iLite effector cells V-variant (1.2 × 10⁵ cells/well) or NFAT responsive effector cells V-variant cells and EGFR⁺ HEK293 target cells were mixed at an E:T ratio of 4 : 1 and incubated for 6 at 37°C in the presence of increasing concentrations of cetuximab 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 fold induction relative to the control without cetuximab.

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