Strategy of macromolecular grafting onto a gold substrate dedicated to protein-protein interaction measurements.
Fiche publication
Date publication
mars 2006
Journal
Biosensors & bioelectronics
Auteurs
Membres identifiés du Cancéropôle Est :
Dr BOIREAU Wilfrid, Pr DELAGE-MOURROUX Régis
Tous les auteurs :
Mansuy-Schlick V, Delage-Mourroux R, Jouvenot M, Boireau W
Lien Pubmed
Résumé
Many biotechnology applications use proteins immobilized on surface. For biosensor, the sensing layer is a key component interfacing the transducer and the sample. Strategies employed to activate the bidimensional surface act directly on the performance of the biosensor. In this paper we propose a novel strategy for engineered proteins self-assembly. Our original supramolecular structure allows a direct and fast covalent attachment of proteins onto bare gold substrate through a homobifunctional cross-linker, 1,4-di-([2'-pyridyldithio]propionamido)butane (DPDPB). In this work, engineered proteins and linker-protein complexes were synthesized and characterized by gel electrophoresis, chromatography and spectroscopy experiments. Macromolecular construction "DPDPB-GST tag-GEC1 protein" was conceived in order to guarantee a 2D architecture enhancing the capabilities of the target (tubulin) to recognize its partner (GEC1). Surface plasmon resonance measurements clearly showed potential of this particular self-assembled protein layer compared to a commercial immunosensor interface. At the concentrations tested, the recognition process occurs between tubulin and the immobilized GEC1; moreover enhanced binding was obtained with the home-made 2D sensing layer more than with 3D carboxymethyl dextran matrix.
Mots clés
Biosensing Techniques, instrumentation, Coated Materials, Biocompatible, chemistry, Crystallization, methods, Feasibility Studies, Gold, chemistry, Immunoassay, methods, Macromolecular Substances, chemistry, Protein Interaction Mapping, methods, Surface Plasmon Resonance, methods
Référence
Biosens Bioelectron. 2006 Mar;21(9):1830-7