Functionalization of 3-D Structures for Grafting of Biological Molecules
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Functionalization of 3-D Structures for Grafting of Biological Molecules Cécile Oillic1, Pierre Mur1, Elisabeth Blanquet2, Guillaume Delapierre1, Françoise Vinet1 and Thierry Billon1 1 CEA-DRT-LETI-CEA/GRE, 17 rue des Martyrs 38054 Grenoble Cedex9, France 2 LTPCM/ ENSEEG/ INPG, BP 75 1130 rue de la piscine, 38402 Saint Martin d’Hères cedex, France ABSTRACT Even though most microarrays present good quality, accuracy and reliability, they are made on a planar surface structure, which neither enough increases the accessibility of the targets to the probes nor the loading capacity of the solid support. To achieve a high density of reactive functions, the use of a non-planar structure is investigated to increase the available surface area for grafting of biomolecules. We propose to build up a pseudo-three-dimensional silicon structure, covered with a specific oxide layer, and then functionalized, allowing to introduce covalent and stable bindings of amino-modified oligonucleotides probes on the reactive layer of the support. The performances of these supports after silanisation are investigated by means of hybridization experiments using complementary fluorescent labeled-oligonucleotides targets. Our results indicate that these novel surfaces provide a higher specific surface area for attaching biomolecules and higher accessibility of the targets, which will increase the density of biomolecules and hence, the sensitivity of the fluorescence signal in comparison to the results obtained with a planar surface structure.
INTRODUCTION DNA microarrays are considered as powerful tools, allowing massively parallel analysis of biological processes and therefore a reduction of time analysis [1]. Thin films materials with specific surface properties are useful for these applications, in particular for covalent immobilization of various biological molecules such as nucleic acids and proteins. A high number of different molecular probes immobilized at well defined locations on a solid support allow simultaneous detection of fluorescently labeled biomolecules (called targets). After biological recognition between probes and targets, the microarray is analyzed by the measurement of the fluorescence image of its surface. Due to the low amount of biological samples, a main issue is to detect fluorescent targets with the lowest detection threshold and the best sensitivity. Different strategies for improving the sensitivity of fluorescent detection have already been investigated, for example, by increasing the hybridization events onto the microarray surface [2]. Indeed, even though most microarrays present good characteristics, they are made on a planar surface structure, which limits the accessibility of the targets to the probes and the loading capacity. A solution for this issue consists in increasing the specific surface area and hence the probes grafting density using silicon nanostructures.
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EXPERIMENTAL DETAILS Elaboration of the 3-D support The deposition of the silicon nanostructures is performed o
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