In this paper, the composite films of Nafion/Hb/FDU-Pt-15/PDDA were constructed by layer-by-layer assembly technique. The FDU-15-Pt mesoporous materials were synthesized and characterized by transmission electron microscopy (TEM) and X-ray diffraction (XRD). The resulting films were verified by electrochemical impedance spectroscopy (EIS), cyclic voltammetry and UV-vis spectroscopy. The direct electrochemical and electrocatalytic properties of hemoglobin (Hb) in the Nafion/Hb/FDU-15-Pt/PDDA films were also investigated. A pair of stable and well-defined reversible redox peaks was observed and the formal potential of Hb Fe(III)/Fe(II) redox couple was found to be -0.324 V (vs. SCE). Based on a series of optimized conditions, the films modified electrode displayed a good electrocatalytic activity to the reduction of hydrogen peroxide (H2O2), which had linear current response from 2.0 x 10(-6) M to 6.0 x 10(-2) M with the detection limit of 1.0 x 10(-6) M (S/N=3). The apparent Michaelis-Menten constant (K(m)(app)) was determined to be 0.15 mM in the range of lower concentration of H2O2. The apparent heterogeneous electron transfer rate constant (k(s)) was 1.05 +/- 0.03 s(-1). The good sensitivity, reproducibility, excellent stability and wide linear range make Nafion/Hb/FDU-15-Pt/PDDA a promising application of the preparation of third-generation biosensor.
2009 Elsevier B.V. All rights reserved.