Surface characterization and efficiency of a matrix-free and flat carboxylated gold sensor chip for surface plasmon resonance (SPR) |
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Authors: | L Roussille G Brotons L Ballut G Louarn D Ausserré S Ricard-Blum |
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Institution: | 1.Laboratoire de Physique de l’Etat Condensé,UMR 6087 CNRS-Université du Maine, molecular landscape and biophotonic horizons,Le Mans,France;2.Institut de Biologie et Chimie des Protéines,UMR 5086 CNRS-Université Lyon 1, IFR 128 Biosciences Lyon-Gerland,Lyon Cedex 07,France;3.Institut des Matériaux Jean Rouxel,UMR 6502 CNRS-Université de Nantes,Nantes,France |
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Abstract: | We report the preparation and characterization of a matrix-free carboxylated surface plasmon resonance (SPR) sensor chip with
high sensing efficiency by functionalizing a bare gold thin film with a self-assembled monolayer of 16-mercaptohexadecanoic
acid (SAM–MHDA chip). The self assembled monolayer surface coverage of the gold layer was carefully evaluated and the SAM
was characterized by infrared reflection absorption spectroscopy, X-ray photoemission spectroscopy, atomic force microscopy,
X-ray reflectivity-diffraction, and SPR experiments with bovine serum albumin. We compared the SPR signal obtained on this
chip made of a dense monolayer of carboxylic acid groups with commercially available carboxylated sensor chips built on the
same gold substrate, a matrix-free C1 chip, and a CM5 chip with a ~100 nm dextran hydrogel matrix (GE Healthcare). Two well-studied
interaction types were tested, the binding of a biotinylated antibody (immunoglobulin G) to streptavidin and an antigen–antibody
interaction. For both interactions, the well characterized densely functionalized SAM–MHDA chip gave a high signal-to-noise
ratio and showed a gain in the availability of immobilized ligands for their partners injected in buffer flow. It thus compared
favourably with commercially available sensor chips. |
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