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. 2014 Nov 18;107(10):2456–2466. doi: 10.1016/j.bpj.2014.10.007

Figure 4.

Figure 4

The linker length and IFNα binding affinity modulate the targeted cell signaling activation by antibody-cytokine complexes. K562 (nontarget) and K562 CD20+ (target) cells were incubated with DNA-protein complexes (as shown in Fig. 1A, with three different DNA linker lengths) for 30 min, lysed, and assayed for Stat1 phosphorylation by ELISA. Simulations were performed using the two-component approach described in Fig. 1 to model the experimental results. (A and B) Stat1 phosphorylation curves are shown for DNA-protein complexes incorporating (A) IFNα(R144A)-SNAP and (B) IFNα(WT)-SNAP. (C and D) EC50 values were calculated and compared for the experimental and simulated data by fitting a four-parameter logistic function to the curves for the (C) IFNα(R144A)-SNAP and (D) IFNα(WT)-SNAP complexes. To see this figure in color, go online.