Molecular hydrogel-stabilized enzyme with facilitated electron transfer for determination of H2O2 released from live...

In this work, small molecular hydrogel was first employed as a surrounding matrix to stabilize an enzyme model, Cytochrome c (Cyt c), and more importantly to facilitate electron transfer between redox enzyme and electrode. Direct electron transfer of Cyt c was successfully achieved in the molecular hydrogel with redox formal potential (E(0')) of 100.7 ± 3.2 mV versus Ag|AgCl and heterogeneous electron transfer rate constant (ks) up to 18.6 ± 2.3 s(-1). Experimental data demonstrated that Cyt c was stably immobilized into the molecular hydrogel and retained its inherent bioactive activity toward H2O2. The direct redox reaction of Cyt c, followed by the biochemical reaction between Cyt c and H2O2, established a reliable approach to determine H2O2 at an optimized potential with high selectivity over other reactive oxygen species (ROS), oxygen, metal ions, ascobic acid (AA), and so on. In addition, the present biosensor for H2O2 also exhibited wide linear range and low detection limit, which fulfills the requirements for detection of H2O2 in a biological system. The remarkable analytical performance of the present biosensor, as well as the long-term stability and good reproducibility ascribed to the molecular hydrogel-stabilized enzyme, provided a durable platform for real-time determination of H2O2 from live cells.