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. 2014 Dec 17;4:7518. doi: 10.1038/srep07518

Figure 4. Characterization of affinity matured VEGFR2-binding Affibody molecules selected by staphylococcal display.

Figure 4

(a) Representative sensorgrams from SPR analysis of affinity-matured Affibody molecules (ZVEGFR2_22 and ZVEGFR2_40) binding to immobilized VEGFR2, showing the response signal. Affibody molecules were injected at concentrations of 5, 10 and 20 nM. Data is referenced by subtraction of simultaneous responses from reference surface. The experiment was performed in duplicates. (b) Representative results from SPR-based competition assay. Sensorgrams were obtained from a double injection, where 1 μM of ZVEGFR2_22 was injected (1), immediately followed by a second injection (2) of either a combination of 1 μM of ZVEGFR2_22 and 1 μM of ZVEGFR2_40, or 2 μM of ZVEGFR2_22, over immobilized human or murine VEGFR2. The experiment was performed in duplicates. (c) Representative results from SPR-based assay of VEGF blocking. 40 nM of human VEGFR2, which had been pre-incubated for 40 min with a 25 × molar excess of ZVEGFR2_22, was injected over a surface of immobilized human VEGF-A. The experiment was performed in duplicates.