Hydrogen peroxide sensor based on horseradish peroxidase immobilized on an electrode modified with DNA-L-cysteine-gold-platinum nanoparticles in polypyrrole film

A third-generation mediator-free hydrogen peroxide biosensor was fabricated by combining electrodeposition and self-assembly techniques. In this strategy, Au-Pt hybrid nanoparticles were electrodeposited on a film of polypyrrole, and L-cysteine was assembled to their surface by exploiting the strong affinity between mercapto groups and nanoparticles. Then, DNA and horseradish peroxidase were respectively self-assembled on the surface of the electrode. The resulting electrode was characterized by impedance spectroscopy, and the electrode surface (without enzyme) was characterized by scanning electron microscopy. The response of the sensor towards hydrogen peroxide, as investigated by cyclic voltammetry and chronoamperometry, is linear between 4.9 µM to 4.8 mM, with a detection limit of 1.3 µM (at an S/N of 3). The apparent Michaelis-Menten constant $ \left( {K_M^{app}} \right) $ is 0.69 mM.