Figure 3.

Cell-based ADCC assays of Fc variants. (A) Reporter cell-based assay for Fc variants. Jurkat-NFAT-FcγRIIIa reporter cell was used for monitoring the early nuclear translocation of NFAT upon ADCC induction. The reporter cells were stimulated by 1 µg well-coated Fc variants for 24 h and bioluminescent signals were measured. (B) PBMC-based ADCC assay of antibody variants. SK-BR-3 target cells were co-cultured with human PBMCs from 3 healthy donors in the presence of 1 µg/mL trastuzumab or its Fc variants. Target cell lysis was measured by the release of lactate dehydrogenase (LDH). Each color represents a different donor. (C) Concentration-dependent study of trastuzumab or its Fc variants-mediated ADCC on SK-BR-3 cells using PBMC as effector cells. (D) Concentration-dependent study of rituximab or its Fc variants-mediated ADCC on Ramos cells using PBMC as effector cells. Bars represent the mean ± SEM of three independent experiments.