Journal
BIOSENSORS & BIOELECTRONICS
Volume 22, Issue 2, Pages 170-177Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2005.12.013
Keywords
DNA immobilization; rehybridisation; affinity biosensors; nanocrystalline diamond; fatty acids; EDC coupling; amide bond formation
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Chemical vapour deposited (CVD) diamond is a very promising material for biosensor fabrication owing both to its chemical inertness and the ability to make it electrical semiconducting that allows for connection with integrated circuits. For biosensor construction, a biochemical method to immobilize nucleic acids to a diamond surface has been developed. Nanocrystalline diamond is grown using microwave plasma-enhanced chemical vapour deposition (MPECVD). After hydrogenation of the surface, 10-undecenoic acid, an w-unsaturated fatty acid, is tethered by 254nm photochemical attachment. This is followed by 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide (EDC)-mediated attachment of amino, (NH(2))-modified dsDNA. The functionality of the covalently bound dsDNA molecules is confirmed by fluorescence measurements, PCR and gel electrophoresis during 35 denaturation and rehybridisation steps. The linking method after the fatty acid attachment can easily be applied to other biomolecules like antibodies and enzymes. (c) 2006 Elsevier B.V. All rights reserved.
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