ABSTRACT
Through alternative adsorption of dextran sulfate (DS) and mesoporous SnO2 (Meso-SnO2), {Meso-SnO2/DS}n films were constructed and applied for the immobilization of hemoglobin (Hb). For comparison, the same-sized non-mesoporous SnO2 nanopraticles (Nonmeso-SnO2) based films were also prepared. Thus-formed films were characterized by many techniques in detail. The electrochemical impedance spectroscopy (EIS, using ferricyanide/ferrocyanide as a redox probe) experiments confirmed the charge transfer resistance increased linearly with the number of Meso-SnO2/DS bilayers, which was smaller than that of Nonmeso-SnO2/DS bilayers. Further EIS, UV-vis and AFM experiments indicate that Meso-SnO2/DS film exhibit enhanced immobilization ability of Hb with favorable orientation in comparison with that of Nonmeso-SnO2/DS film. The adsorbed Hb at Meso-SnO2/DS film showed a good direct electron transfer behavior with the heterogeneous electron transfer rate constant (ks) of 8.89 +/-0.32 s(-1), and excellent electrocatalysis to the reduction of O2 and H2O2 in pH 7.0 buffers.
Subject(s)
Biosensing Techniques/methods , Hemoglobins/chemistry , Hemoglobins/metabolism , Nanoparticles/chemistry , Tin Compounds/chemical synthesis , Adsorption , Catalysis , Electrochemistry , Hemoglobins/ultrastructure , Humans , Immobilized Proteins/chemistry , Immobilized Proteins/metabolism , Immobilized Proteins/ultrastructure , Nanoparticles/ultrastructure , Tin Compounds/chemistryABSTRACT
Direct electrochemistry of hemoglobin (Hb) was observed at glassy carbon electrode (GCE) modified with undoped nanocrystalline diamond (UND) and Hb multilayer films via layer-by-layer assembly. UV-VIS absorbance spectroscopy, electrochemical impedance spectroscopy and cyclic voltammograms were employed to characterize the film. The results showed that the UND had the effect of enhancing the electron transfer between Hb and the electrode surface. Hb in the multilayer films maintained its bioactivity and structure. It also exhibited a good catalytic activity towards the reduction of H(2)O(2). The reciprocal of catalytic current showed a linear dependence on the reciprocal of H(2)O(2) concentration ranging from 0.5 microM to 0.25 mM with a detection limit of 0.4 microM. The apparent Michaelis-Menten constant was estimated to be 0.019 mM.
