Using a mixed self-assembly method, this research utilized modified Mb-functionalized gold nanoparticles (Mb-AuNPs-MPA) on the gold electrode surface to prepare a biosensor which was applied to detect superoxide anion-free electron mediators. Together with the study on the performance of the sensor, the characteristics of modified nanoclusters were investigated by UV-Vis and FTIR and the electrochemical characteristics of the electrode surface were investigated using electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV), respectively. It was demonstrated that the charge transfer resistance (Rct) of the modified electrode (Mb-MPA-AuNPs/Au) was 7862 Ω, the exchange current density () was 32.7 A/cm. And a pair of reversible and symmetrical redox peaks appeared after the coordination of Cu, and the electrical signal response of Cu/Mb-AuNPs-MPA/Au reached the highest. The superoxide anion generated in the basic DMSO system was determined by CV using the modified electrode Cu/Mb-AuNPs-MPA/Au. It was discovered that the superoxide anion had a strong disproportionation effect.
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