Figure 1.

Binding properties of anti-mesothelin antibody and CAR constructs on primary human T cells. (A) Mesothelin expression in the established cell lines. Cell extracts from mesothelin-transfected cells were subjected to Western blot analysis. The blot was incubated with a mouse monoclonal antibody mesothelin (K1). GAPDH was used as a loading control. (B) The mean fluorescence intensity (MFI) of different scFv proteins bound cells was determined by flow cytometric analysis. (C) Binding specificity of the anti-mesothelin antibody to MSLN-transfected CHO-K1 or PANC-1 cells. (D) Affinity measurements of antibody binding to mesothelin through Biacore Surface Plasmon resonance. (E) A schematic diagram showing the MSLN-specific CAR used in this study. (F) The primary human T cells efficiently express MSLN-specific CAR, as measured by flow cytometry. Mock T cells were demonstrated by assessing the expression of eGFP