It is always challenging to construct a smart functional nanostructure with specific physicochemical properties to real time detect biointeresting molecules released from live-cells. We report here a new approach to build a free-standing biomimetic sensor by covalently bonding RGD-peptide on the surface of pyrenebutyric acid functionalized graphene film. The resulted graphene biofilm sensor comprises a well-packed layered nanostructure, in which the RGD-peptide component provides desired biomimetic properties for superior human cell attachment and growth on the film surface to allow real-time detection of nitric oxide, an important signal yet short-life molecule released from the attached human endothelial cells under drug stimulations. The film sensor exhibits good flexibility and stability by retaining its original response after 45 bending/relaxing cycles and high reproducibility from its almost unchanged current responses after 15 repeated measurements, while possessing high sensitivity, good selectivity against interferences often existing in biological systems, and demonstrating real time quantitative detection capability toward nitric oxide molecule released from living cells. This study not only demonstrates a facial approach to fabricate a smart nanostructured graphene-based functional biofilm, but also provides a powerful and reliable platform to the real-time study of biointeresting molecules released from living cells, thus rendering potential broad applications in neuroscience, screening drug therapy effect, and live-cell assays.
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