Binding and internalization of bispecific immunotoxins.

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Binding and internalization of bispecific immunotoxins. Binding and internalization of bispecific immunotoxins. Increased concentrations of FITC-labeled DT23EpCAM and DTEpCAM23 were incubated with (A) HT-29 and (B) SK-BR-3 cells for 45 min. Following FACS, the MFI was plotted versus immunotoxin concentration. C, internalization efficiency of bound immunotoxin was determined by incubating cells with bound FITC-labeled DT23EpCAM and DTEpCAM23 at 37°C. Internalized protein was differentiated from surface bound by quenching extracellular FITC fluorescence with trypan blue. Columns, percentage of initially bound protein internalized. Brad J. Stish et al. Clin Cancer Res 2007;13:3058-3067 ©2007 by American Association for Cancer Research