Fig. 3

Glycoengineered anti-MCSP antibody LC007 synergises with CD16-158V-CAR for effective melanoma cell recognition and lysis. a Schematic overview of constructs used in this figure; the anti-MCSP antibody (LC007 or glycoengineered LC007GE), CD16-CAR with VH, FL, VL variants and Vdel. b CD16 VH-CAR-, CD16 VL-CAR-, CD16 FL-CAR-, CD16-del-transduced T cells were co-cultured for 48 h with 20.000 MCSP⁺ A375 melanoma cells in the presence or absence of either 10 µg/ml of the anti-EGFR antibody cetuximab (positive control), increasing doses of the anti-MCSP antibodies LC007 or increasing doses of the glycoengineered LC007GE (10, 1, and 0.1 µg/ml) as indicated in the figure. IFN-γ production was measured by ELISA. c 10⁵ CD16 VH-CAR-, CD16 VL-CAR-, CD16 FL-CAR- or CD16-del-transduced T cells were added after overnight incubation to 10⁴ MCSP⁺ A375 melanoma cells, in the presence or absence of either 10 µg/ml of the anti-EGFR antibody cetuximab, 10 µg/ml of the anti-MCSP antibody LC007 or glycoengineered LC007GE, as indicated in the figure. The time point when T cells and antibodies were added to the system are indicated by the black arrows. Cytotoxicity was measured in real-time, as a mean of cell viability by xCELLigence technology. All graphs show mean values of at least three technical replicate and each experiment shown is a representative figure of at least three independent experiments. For figure B a two-sided unpaired Student´s t test was used and for C a two-way ANOVA with Bonferroni-correction was used to determine the p-values. A p-value < 0.05 was considered statistically significant