Factors that Affect Protein Adsorption on Nanostructured Titania Films. A Novel Spectroelectrochemical Application to Sensing
- 9 November 2001
- journal article
- research article
- Published by American Chemical Society (ACS) in Langmuir
- Vol. 17 (25), 7899-7906
- https://doi.org/10.1021/la010309b
Abstract
Nanoporous, thick (8 μm) films of titania (TiO2) were prepared and used for the immobilization of proteins. A detailed study has been made into the factors influencing protein adsorption on TiO2. Among these, we investigated pH, ionic strength of solution, protein surface charge, protein size, and immobilization time. Protein immobilization is found to be remarkably stable, attributed to secondary binding processes occurring after the initial immobilization. We also investigated the electrochemical properties of these films using cyclic voltammetry and spectroelectrochemistry and found that not only was direct reduction of the FeIII−heme to FeII−heme of both cytochrome-c and hemoglobin possible but that all the protein in the film is electroactive. We further demonstrate the use of a hemoglobin/TiO2 film as an aerobic sensor for nitric oxide. Optical sensing is demonstrated, with a limit of detection of 1 μM.Keywords
This publication has 32 references indexed in Scilit:
- Application of chemistry to in vitro diagnostic testsChemical Society Reviews, 2000
- Effect of Trehalose on Alkaline Transition of Cytochrome-cThe Journal of Physical Chemistry B, 2000
- Interruption and Time-Resolution of Catalysis by a Flavoenzyme Using Fast Scan Protein Film VoltammetryJournal of the American Chemical Society, 2000
- Photoelectrochromic heterosupramolecular assembliesJournal of Materials Chemistry, 2000
- Catalytic Electrochemical Synthesis Using Nanocrystalline Titanium Dioxide Cathodes in MicroemulsionsLangmuir, 1998
- Chronoabsorptometric determination of adsorption isotherms for cytochrome c on tin oxide electrodesLangmuir, 1992
- Interfacial electron transfer of flavin coenzymes and riboflavin adsorbed on textured TiO2 filmsBioelectrochemistry and Bioenergetics, 1991
- Light-induced reduction of cytochrome c by colloidal TiO2Bioelectrochemistry and Bioenergetics, 1986
- Bacterial proteins with co-binding b- or c-type haem fucntions and absorption spectroscopyBiochimica et Biophysica Acta (BBA) - Reviews on Bioenergetics, 1984
- Haem exposure as the determinate of oxidation–reduction potential of haem proteinsNature, 1978