EDC-mediated DNA attachment to nanocrystalline CVD diamond films

被引:61
作者
Christiaens, P.
Vermeeren, V.
Wenmackers, S.
Daenen, M.
Haenen, K.
Nesladek, M.
vandeVen, M.
Ameloot, M.
Michiels, L.
Wagner, P.
机构
[1] Hasselt Univ, Mat Res Inst, B-3590 Diepenbeek, Belgium
[2] Transnatl Univ Limburg, B-3590 Diepenbeek, Belgium
[3] Hasselt Univ, Inst Biomed Res, Sch Life Sci, B-3590 Diepenbeek, Belgium
[4] IMEC, Div IMOMEC, B-3590 Diepenbeek, Belgium
[5] CEA, LTD, SSTM, DETECS,LIST, F-91191 Gif Sur Yvette, France
关键词
DNA immobilization; rehybridisation; affinity biosensors; nanocrystalline diamond; fatty acids; EDC coupling; amide bond formation;
D O I
10.1016/j.bios.2005.12.013
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
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.
引用
收藏
页码:170 / 177
页数:8
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