Fig. 3

Binding of αSTn and αSTn-IgG4 TMs to STn-expressing cancer cells. a Both MDA-MB-231 and MCR cancer cells expressing STn were stained with 1 μg of TMs, mAb 5B9 and PE-labeled anti-mouse-IgG mAb. Staining with αSTn mAbs L2A5 and αSTn TM served as positive controls. L2A5 mAb was detected using an Alexa Fluor 488 anti-mouse IgM mAb. Stained cells (black lines) and respective controls (grey lines) are displayed as histograms. MFI and percentage of positively stained cells (in parenthesis) are shown. Results for one representative binding assay are depicted. b and c Binding affinity curves of αSTn (grey) or αSTn-IgG4 (black) TMs to MDA-MB-231 STn⁺(b) and MCR STn⁺(c) cells were obtained using increasing concentrations of TM. Binding was detected as described above. Data are presented as MFI average values ± SD from four independent experiments