Institut de Recherche Interdisciplinaire (IRI, USR 3078), Parc de la Haute Borne, 50 Avenue de Halley, BP 70478, 59658 Villeneuve d'Ascq, France.
Langmuir. 2010 Jan 19;26(2):1075-80. doi: 10.1021/la903012v.
The successful covalent linking of green fluorescence protein and streptavidin to patterned benzophenone-modified boron-doped diamond (BDD) electrodes is demonstrated. Photoreactive benzophenone moieties were covalently grafted to oxidized diamond surfaces via an esterification reaction. Patterned BDD surfaces were obtained using a UV/ozone lithographic approach either on hydrogen-terminated BDD or on poly(ethylene)-glycol-modified BDD surfaces. UV light (lambda = 365 nm) irradiation of the patterned BDD surfaces in the presence of green fluorescence protein (GFP) or streptavidin resulted in the covalent immobilization of the proteins. The presence of poly(ethylene) glycol chains reduces significantly the nonspecific adsorption of the proteins. The success of the photoimmobilization of streptavidin was evidenced through biomolecular interaction with avidin. The preservation of the biological activity was furthermore underlined by photoimmobilization of peptides directly onto benzophenone modified BDD using a photomask.
成功地将绿色荧光蛋白和链霉亲和素共价连接到图案化的苯并二酮修饰的硼掺杂金刚石（BDD）电极上。通过酯化反应将光反应性苯并二酮部分共价接枝到氧化金刚石表面上。通过 UV/臭氧光刻法在氢终止的 BDD 或聚（乙二醇）修饰的 BDD 表面上获得图案化的 BDD 表面。在存在绿色荧光蛋白（GFP）或链霉亲和素的情况下，用 UV 光（lambda = 365nm）照射图案化的 BDD 表面，导致蛋白质的共价固定。聚乙二醇链的存在显著降低了蛋白质的非特异性吸附。通过与亲和素的生物分子相互作用，证明了链霉亲和素的光固定的成功。通过使用光掩模将肽直接固定到苯并二酮修饰的 BDD 上，进一步强调了生物活性的保留。
