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On the use of aptamer microarrays as a platform for the exploration of human prothrombin/thrombin conversion
Authors:Camille Daniel  Yoann Roupioz  Thierry Livache  Arnaud Buhot
Affiliation:1. Université Grenoble Alpes, INAC–SPrAM, F-38000 Grenoble, France;2. Centre National de la Recherche Scientifique (CNRS), SPrAM, F-38000 Grenoble, France;3. Centre d’Etudes Atomiques (CEA), INAC–SPrAM, F-38000 Grenoble, France
Abstract:Microarrays are particular biosensors with multiple grafted probes that are generally used for parallel and simultaneous detection of various targets. In this study, we used microarrays with aptamer probes in order to follow up the different biomolecular interactions of a single enzyme, the thrombin protein, involved in the complex coagulation cascade. More precisely, thanks to label-free surface plasmon resonance imaging, we were able to monitor in real time an important step in the firing of the coagulation cascade in situ—the enzymatic transformation of prothrombin into thrombin, catalyzed by factor Xa. We were also able to appraise the influence of other biochemical factors and their corresponding inhibiting or enhancing behaviors on thrombin activation. Our study opens the door for the development of a complete microarray-based platform not only for the whole coagulation cascade analysis but also for novel drug screening assays in pharmacology.
Keywords:Aptamers   Thrombin   Prothrombin   Microarrays   Surface plasmon resonance imaging   Coagulation cascade
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