A novel superoxide anion (O(2)(-)) biosensor is proposed based on the immobilization of copper-zinc superoxide dismutase (SOD) in a gold nanoparticle-chitosan-ionic liquid (GNPs-CS-IL) biocomposite film. The SOD-based biosensor was constructed by one-step ultrasonic electrodeposition of GNP-CS-IL composite onto glassy carbon electrode (GCE), followed by immobilization of SOD on the modified electrode. Surface morphologies of a set of representative films were characterized by scanning electron microscopy. The electrochemical performance of the biosensor was evaluated by cyclic voltammetry and chronoamperometry. A pair of quasi-reversible redox peaks of SOD with a formal potential of 0.257V was observed at SOD/GNPs-CS-IL/GCE in phosphate buffer solution (PBS, 0.1M, pH 7.0). The effects of varying test conditions on the electrochemical behavior of the biosensor were investigated. Furthermore, several electrochemical parameters were calculated in detail. Based on the biomolecule recognition of the specific reactivity of SOD toward O(2)(-), the developed biosensor exhibited a fast amperometric response (<5s), wide linear range (5.6-2.7×10(3)nM), low detection limit (1.7nM), and excellent selectivity for the real-time measurement of O(2)(-). The proposed method is promising for estimating quantitatively the dynamic changes of O(2)(-) in biological systems.
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