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Inspired by the high electrocatalytic activity of electrochemically reduced graphene oxide-Au nanoparticles (ERGO-AuNPs) nanohybrid and their strong adsorption capacity for immobilizing biomolecules, we developed an ultrasensitive superoxide anion (O2 •− ) electrochemical biosensor based on hemin immobilized on ERGO-AuNPs nanohybrid modified glass carbon electrode (GCE). The electrochemical properties of immobilized hemin were investigated. A considerably higher surface concentration (Г*) of the immobilized hemin was measured on the order of 1.02×10 −7 mol cm −2 . The results indicated that the ERGO-AuNPs nanohybrid enhanced the electrocatalytic activity of hemin for O2 •− detection due to the synergy generated by ERGO and AuNPs. The resulting biosensor exhibited an excellent amperometric response toward O2 •− , at linear ranges of 3.28×10 −5 to 3.28×10 −4 M with sensitivities of 186 nA μM -1 cm -2 and 124 nA μM -1 cm -2 at 0.5 V and −0.1 V, separately. The apparent Michaelis–Menten constant (Km) was estimated to be 0.51 mM and 0.48 mM at 0.5 V and −0.1 V, respectively, which indicated a clearly stronger interaction of hemin with O2 •− . The results indicated that the biosensor have strong potential for analysis of biological molecules.

Enhanced Electrocatalytic Activity of Graphene-Gold Nanoparticles as a Superoxide Anion Biosensor Based on Immobilized Hemin