Research Article

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

Figure 1

Activation of transcription factors implicated in CD16 signaling. Jurkat cells were cotransfected with a CD16a expression vector, the NL normalization gene, and a reporter gene construct for one of the transcription factors indicated in the figure and incubated in the presence of an equal concentration of wild-type Raji cells and 500 ng/ml of rituximab for 5 hours prior to the addition of Nano-Glo Dual-luciferase reagent (Promega, Madison, WI) and the sequential determination of FL and NL activity. Fold induction was calculated from the normalized FL RLU values observed in the presence of the Jurkat effector cells, Raji target cells, and rituximab after subtraction of the normalized FL RLU values obtained with Jurkat and Raji cells in the absence of rituximab.