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. 2013 Nov 15;8(11):e81919. doi: 10.1371/journal.pone.0081919

Figure 2. Binding of IL12-SS1 (Fv) to MSLN and cancer cells.

Figure 2

(A) The binding specificity of IL12-SS1 (Fv) on MSLN was detected by ELISA. (B) FACS analysis of IL12-SS1 (Fv) on mesothelin-expressing cancer cells. NCI-H226, OVCAR-3, H9, A431, and HEK-293 cells were stained with IL12-SS1 (Fv) which was detected by PE-conjugated anti-histidine antibody (solid line). Mesothelin expression was detected by a mouse anti-MSLN antibody (MN, dotted line) or an irrelevant isotype mAb control (gray surface). Samples were analyzed by flow cytometry. H9: the transfected A431 cell line stably expressing mesothelin; OVCAR-3: human ovarian cancer cell lines; NCI-H226: human mesothelioma cell lines; HEK-293: human embryonic Kidney cell lines. (C) FACS analysis of IL12-SS1 (Fv) on A431 cells. A431 cells were stained with IL12-SS1 (Fv) in the presence of (dotted line) or the absence of (solid line) an anti-IL12R β2 blocking antibody. The binding of IL12-SS1 (Fv) was detected by FITC-conjugated anti-Flag antibody. (D) SS1P and IL12-SS1 (Fv) bound to H9 cells in a dose-dependent manner. H9 cells were stained with indicated concentrations of IL12-SS1 (Fv) and then detected by flow cytometry. The K d value for SS1P and IL12-SS1 (Fv) binding to H9 cells are approximately 2.8 and 60 nM, respectively.