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

Figure 12

Quantification of the ADCC activity of trastuzumab using frozen RTU iLite effector cells and ERBB2⁺ HEK293 target cells or thaw-and-use NFAT effector cells and SK-BR-3 target cells. RTU frozen iLite effector cells V-variant (1.2 × 10⁵ cells/well) or thaw-and-use NFAT effector cells (1.2 × 10⁵ cells/well) and frozen RTU ERBB2⁺ HEK293 target cells or thaw-and-use NFAT effector cells (0.75 × 10⁵ cells/well) and SK-BR-3 target cells at an E:T ratio of 4 : 1 and 6 : 1, respectively, were 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) or Bio Glo, respectively, and the sequential determination of FL and NL activity or FL activity alone. Results are expressed as fold induction relative to the control sample without trastuzumab.