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. 2018 Mar 30;9(24):17117–17132. doi: 10.18632/oncotarget.24965

Figure 1.

Figure 1

(A-C) In vitro characterization of DFO-nimotuzumab kit formulation by biolayer interferometry (BLI) and flow cytometry. (A) BLI binding curves for nimotuzumab (i) and DFO-nimotuzumab (ii) at different concentrations (55.5, 166.7 and 500 nM). A KD of 23 ± 3 nM) and 48 ± 9 nM was obtained for nimotuzumab and DFO-nimotuzumab, respectively. Solid black line indicates raw data, solid red line indicates curve fit, vertical dashed red line indicates transition from association to dissociation phase. (B) Flow cytometry results of nimotuzumab in control MDA-MB-435 (i) with very low levels of EGFR and corresponding EGFR-positive A431 cells (ii and iii). No substantial binding was seen in MDA-MB-435 cells. Binding of DFO-nimotuzumab (iii) in A431 cells was comparable with nimotuzumab (ii). (C) Nimotuzumab (i) and DFO-nimotuzumab (ii) showed saturation binding by flow cytometry with a KD of 13 ± 2 nM and 17 ± 4 nM, respectively.