A third-generation hydrogen peroxide biosensor based on horseradish peroxidase immobilized in a tetrathiafulvalene–tetracyanoquinodimethane/multiwalled carbon nanotubes film

Abstract

A new third-generation biosensor for H2O2 assay was developed on the basis of the immobilization of horseradish peroxidase (HRP) in a nanocomposite film of tetrathiafulvalene–tetracyanoquinodimethane (TTF–TCNQ)/multiwalled carbon nanotubes (MWCNTs) modified gold electrode. The prepared HRP/TTF–TCNQ/MWCNTs/Au electrode was used for the bioelectrocatalytic reduction of H2O2, with a linear range from 0.005 to 1.05mM and a detection limit of 0.5μM for amperometric sensing of H2O2. In addition, a novel method on the basis of electrochemical quartz crystal microbalance (EQCM) measurements was proposed to determine the effective enzymatic specific activity (ESA) of the immobilized HRP for the first time, and the ESA was found to be greater at the TTF–TCNQ/MWCNTs/Au electrode than that at the MWCNTs/Au or TTF–TCNQ/Au electrode, indicating that the TTF–TCNQ/MWCNTs film is a good HRP-immobilization matrix to achieve the direct electron transfer between the enzyme and the electrode.