Diamond exhibits several special properties, for example good biocompatibility and a large electrochemical potential window, that make it particularly suitable for biofunctionalization and biosensing. Here we show that proteins can be attached covalently to nanocrystalline diamond thin films. Moreover, we show that, although the biomolecules are immobilized at the surface, they are still fully functional and active. Hydrogen-terminated nanocrystalline diamond films were modified by using a photochemical process to generate a surface layer of amino groups, to which proteins were covalently attached. We used green fluorescent protein to reveal the successful coupling directly. After functionalization of nanocrystalline diamond electrodes with the enzyme catalase, a direct electron transfer between the enzyme's redox centre and the diamond electrode was detected. Moreover, the modified electrode was found to be sensitive to hydrogen peroxide. Because of its dual role as a substrate for biofunctionalization and as an electrode, nanocrystalline diamond is a very promising candidate for future biosensor applications.

The optical properties of nanocrystalline diamond films grown from a hydrogen-rich CH4∕H2 gas phase by hot filament chemical vapor deposition, as well as from an argon-rich Ar∕CH4 gas phase by microwave plasma enhanced chemical vapor deposition, are reported. The influence of nitrogen incorporation on the optical absorption is investigated. The diamond films are characterized by photothermal deflection spectroscopy and temperature dependent spectrally resolved photoconductivity. An onset of absorption at about 0.8eV in undoped films is attributed to transitions from π to π states introduced into the band gap by the high amount of sp2 bonded carbon at the grain boundaries. Incorporation of nitrogen leads to a strong absorption in the whole energy spectrum, as a result of the increasing number of sp2 carbon atoms. The effect of surface states has been observed in the high energy region of the spectrum. Transitions to the conduction band tail and photothermal ionization processes account for the observed onset at 4.4eV. Photocurrent quenching at about 3.3eV is observed in the case of samples grown from a hydrogen-rich CH4∕H2 gas phase.

Horseradish peroxidase (HRP) has been immobilized on the surface of functionalized nanocrystalline diamond (NCD) thin films. The structure of the modified NCD surface as well as the electrochemical behavior of the whole system was characterized by impedance spectroscopy and cyclic voltammetry. The proximity of HRP heme groups to the NCD surface allowed direct electron transfer between them, resulting in two separated one-electron-transfer peaks at 0.05 V and 0.29 V vs Ag/AgCl, corresponding to the cathodic and anodic process, respectively. The heterogeneous electron-transfer constant for both processes was calculated to be 0.066 s(-1), the charge-transfer coefficient alpha = 0.49, and the immobilized enzymatic layer about 2.10(-10) mol/cm2. The modified NCD electrode was used as a third-generation biosensor for hydrogen peroxide determination showing a linear response in the 0.1-45 mM H2O2 range, at +0.05 V vs Ag/AgCl.

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