Nanobiologie : la révolution diagnostique

National audienceDe la nanotechnologie ... Les nanotechnologies recouvrent l'ensemble des techniques, outils et procédés qui permettent de manipuler la matière à une échelle en dessous de la centaine de nanomètres (1 nanomètre = 1 milliardième de mètre), d'élaborer de nouveaux matériaux et composants toujours plus petits, de construire atome par atome de nouvelles molécules et d'exploiter leurs propriétés en vue de nouvelles applications. Les nanotechnologies devraient constituer dans les prochaines années un marché considérable. À l'horizon 2015, 15% de l'activité manufacturière mondiale serait concernée par des dispositifs ou des matériaux utilisant des avancées issues des nanotechnologies. En 2008, le montant du marché mondial est estimé à 500 milliards de dollars et pourrait doubler en 2015, selon la National science foundation. Toutefois, les nanotechnologies suscitent également des inquiétudes. Manipuler la matière à l'échelle moléculaire et interférer avec le monde du vivant soulève des questions éthiques et sanitaires qui doivent être prises en considération par les pouvoirs publics et les acteurs concernés. La nanotechnologie peut être considérée comme une évolution logique de la microélectronique dont l'objectif est la fabrication de circuits intégrés, familièrement appelés « puces électroniques » dont la référence en matériau de base est le silicium. A côté de la nanoélectronique industrielle a émergé la nanotechnologie moléculaire. Le but de cette dernière discipline est de pouvoir assurer un contrôle parfait sur la structure de la matière et de pouvoir construire des objets complexes avec une précision à l'échelle de l'atome ou de la molécule. ... aux nanoBIOtechnologies... Un important challenge dans les dispositifs de BIOcapteurs et de diagnostic est de savoir comment reconstituer/étudier des mécanismes biologiques pertinents à la surface d'une biopuce et quels outils analytiques sont convenables pour fournir rapidement des informations justes sur la structure des molécules attachées à la surface. Un meilleur contrôle dans la réalisation de biopuces peut être obtenu en combinant différentes approches interdisciplinaires, avec pour préoccupation de répondre à une question clé biologique. Les recherches menées au sein de la plateforme CLIPP (Clinical & Innovation Proteomic Platform), rassemblant des compétences multidisciplinaires, sont focalisées sur cet objectif. L'ensemble « conception, réalisation et caractérisation » de puces à protéines représente un des axes de recherche de la plateforme CLIPP. Nous développons différentes stratégies d'ingénierie/structuration de matériaux, de fonctionnalisation chimique et d'immobilisation de biomolécules qui conduit à l'optimisation d'évènements de reconnaissance et leur caractérisation à la surface de la puce. Dernièrement, nous avons développé une méthode simple pour la préparation de substrat d'or présentant des terrasses atomiquement planes, et permettant une caractérisation biomoléculaire à l'échelle nanométrique1. Le mode d'action du taxol sur les microtubules et les conséquences sur leur conformation a pu également être élucidé par observation par microscopie à force atomique (AFM) en conditions physiologiques2. Nous avons également développé des approches originales, consistant à combiner une analyse « globale » par Surface Plasmon Resonance (SPR) et un moyen de caractérisation à l'échelle nanométrique (AFM, Spectroscopie de Photo-électrons X, spectrométrie de masse (MS)). Nous avons en particulier démontré que l'orientation d'une protéine immobilisée en monocouche sur une surface, peut être déterminée par la détection de fragments secondaires spécifiques de la protéine par TOF-SIMS3. Ces développements ont contribué à l'établissement d'une nouvelle combinaison « Analyse des interactions Biomoléculaires / spectrométrie de masse (BIA-MS) basée sur une procédure totalement « on the chip »4. Nous proposons une approche bas coût combinant l'analyse sur un BIAcore 2000 sur puces synthétisées par nos soins (réalisation du substrat et greffage d'anticorps) avec l'identification par MS, directement sur la puce et sans étape d'élution. Utilisant cette technique, l'identification de complexes protéiques a pu être obtenue, et permet ainsi de compléter l'approche BIA-MS dans la découverte et l'analyse de complexes protéiques. Dernièrement, ces développements ont été transposés à de l'analyse multiplexée sur micro-arrays. Les premiers résultats de détection de biomarqueurs dans du plasma s'avèrent être très prometteurs pour une approche diagnostic moyen/haut débit

Preparation of flat gold terraces for protein chip developments

International audienceA simple method to prepare flat gold terraces on mica for atomic force microscopy biomolecular characterisation is described. The procedure includes preheating of the substrate, metal deposition and an annealing step. All of these steps are at elevated temperatures (300–420°C). This approach allows one to prepare large flat gold terraces (200– 500 nm), which constitute ideal substrates for visualisation and characterisation of a self-assembly monolayer of biomolecules at the nanoscale. The authors illustrated this potential of characterisation with the reconstitution of a protein monolayer

Straight GDP-Tubulin Protofilaments Form in the Presence of Taxol

International audienceMicrotubules exist in dynamic equilibrium, growing and shrinking by the addition or loss of tubulin dimers from the ends of protofilaments. The hydrolysis of GTP in b-tubulin destabilizes the microtubule lattice by increasing the curvature of protofilaments in the microtubule and putting strain on the lattice. The ob- servation that protofilament curvature depends on GTP hydrolysis suggests that microtubule destabil- izers and stabilizers work by modulating the curvature of the microtubule lattice itself. Indeed, the microtu- bule destabilizer MCAK has been shown to increase the curvature of protofilaments during depolymeriza- tion. Here, we show that the atomic force microscopy (AFM) of individual tubulin protofilaments provides sufficient resolution to allow the imaging of single pro- tofilaments in their native environment. By using this assay, we confirm previous results for the effects of GTP hydrolysis and MCAK on the conformation of pro- tofilaments. We go on to show that taxol stabilizes microtubules by straightening the GDP protofilament and slowing down the transition of protofilaments from straight to a curved configuration

Standardisation de la métrologie des EVs en milieu complexe

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Accent 2"/&gt <w:LsdException Locked="false" Priority="66" Name="Medium List 2 Accent 2"/&gt <w:LsdException Locked="false" Priority="67" Name="Medium Grid 1 Accent 2"/&gt <w:LsdException Locked="false" Priority="68" Name="Medium Grid 2 Accent 2"/&gt <w:LsdException Locked="false" Priority="69" Name="Medium Grid 3 Accent 2"/&gt <w:LsdException Locked="false" Priority="70" Name="Dark List Accent 2"/&gt <w:LsdException Locked="false" Priority="71" Name="Colorful Shading Accent 2"/&gt <w:LsdException Locked="false" Priority="72" Name="Colorful List Accent 2"/&gt <w:LsdException Locked="false" Priority="73" Name="Colorful Grid Accent 2"/&gt <w:LsdException Locked="false" Priority="60" Name="Light Shading Accent 3"/&gt <w:LsdException Locked="false" Priority="61" Name="Light List Accent 3"/&gt <w:LsdException Locked="false" Priority="62" Name="Light Grid Accent 3"/&gt <w:LsdException Locked="false" Priority="63" Name="Medium Shading 1 Accent 3"/&gt <w:LsdException Locked="false" Priority="64" Name="Medium Shading 2 Accent 3"/&gt <w:LsdException Locked="false" Priority="65" Name="Medium List 1 Accent 3"/&gt <w:LsdException Locked="false" Priority="66" Name="Medium List 2 Accent 3"/&gt <w:LsdException Locked="false" Priority="67" Name="Medium Grid 1 Accent 3"/&gt <w:LsdException Locked="false" Priority="68" Name="Medium Grid 2 Accent 3"/&gt <w:LsdException Locked="false" Priority="69" Name="Medium Gr

Immunocapture des vésicules extracellulaires

International audienceImmunocapture des vésicules extracellulaire

Microsystems and NanoBioanalytical tools for characterization of cellular secretome

National audienceMicrosystems and NanoBioanalytical tools for characterization of cellular secretome<br&g

Plateforme NanoBioAnalytique pour la caractérisation des vésicules extracellulaires

International audienceWorkshop « À la conquête des nanovésicules biologiques

Les vésicules extracellulaires: Définition, séparation, caractérisation

International audienceNo abstract availabl

Higher-order Lamb waves with quasi-zero surface displacement components on a GaAs piezoelectric plate

International audienceHigher-order Lamb waves with quasi-zero surface displacement components are reported on (100)-cut GaAs propagating along the &lt;110&gt; direction where the total displacement at the surface of the plate is less than 10% of the maximum total displacement. The dispersion curves and the displacement component profiles show the reduction of total displacement at the surface of the plate starting when the phase velocities of the higher-order modes are crossing the shear bulk acoustic wave velocity to the value as low as 5%. Due to the concentration of acoustic energy inside the plate, the reported quasi-zero plate acoustic waves (QZ-PAW) further reduce the radiation of acoustic energy when the plate surface is in contact with liquid. The experimental results validate the occurrence of QZ-PAW with a reduction of viscous damping insertion loss compared to previously reported quasi-longitudinal Lamb waves. The results demonstrate the potential QZ-PAW mode for emerging applications such as dual-mode PAW sensors, PAW devices with integrated sensor and actuator, thin-film and ultra-high frequency PAW sensors in highly viscous liquid media

Morphological and functional characterization of the consequences of the oxidative stress experimentally induced on skin cell models in vitro

During the skin ageing, the produced radical species make unstable the "proliferation / differentiation / apoptosis" coupling of epidermal cells; we propose to study this loss of cell integrity, by a double original and complementary approach: - the establishment of proteomic comparative profiles, by spectrofluorimetric, 2D chromatographic/mass spectrometry techniques, - the exploration of cell growth and morphology, by a spectroscopic and topographic study by atomic force microscopy. This work will allow identifying the better biomarker(s) of skin ageing modeled in vitro by a chosen chemical treatment using glyphosate. Original molecules with cell protection effect, able to correct the induced morphological and/or functional disorders could also been identified. Our first results revealed cell morphology of a line of keratinocytes grown on different substrates. A glyphosate dose-effect was noted through an important cell destructuration. Preliminary force spectroscopy measurements on cells revealed also specific interactions with glyphosate. These experimental results based on AFM investigation should highlight the first steps of the oxidative stress mechanism, by visualization and quantification of the phenomenon after its induction at the individual cell scale