Fig. 1

Generation and binding characteristics of the NKG2D–CD16/CD3 fusion proteins. a Schematic illustration of the NKG2D-CD3 and NKG2D-CD16 BFP consisting of the extracellular domain of NKG2D (F78-V216) fused with a CH₂-linker to a CD3 or CD16 specific Fab-fragment. b Exemplary results of a size exclusion chromatography (left and middle panel) and a SDS PAGE (right panel) for both BFP. c Specific binding of the BFP was analyzed by flow cytometry using C1R-MICA transfectants and T cells and NK cells of healthy donors incubated with control (open peaks) or the BFP (shaded peaks) followed by an anti-human PE conjugate. d PBMC of healthy donors were incubated with the indicated concentrations of the BFP or NKG2D-Fc-ADCC followed by an anti-human PE conjugate and analyzed by flow cytometry. Exemplary data from one representative experiment of a total of three with similar results are shown. Mean fluorescence intensity (MFI) levels and EC₅₀ were calculated using a sigmoidal dose-response curve with GraphPad Prism