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| - Immobilization of Ligands with Precise Control of Density toElectroactive Surfaces
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| - We report a broadly applicable surface chemistry methodology to immobilize ligands, proteins,and cells to an electroactive substrate with precise control of ligand density. This strategy is based on thecoupling of soluble aminooxy terminated ligands with an electroactive quinone terminated monolayer. Thesurface chemistry product oxime is also redox active but at a different potential and therefore allows forreal−time monitoring of the immobilization reaction. Only the quinone form of the immobilized redox pairis reactive with soluble aminooxy groups, which allows for the determination of the yield of reaction, theability to immobilize multiple ligands at controlled densities, and the in−situ modulation of ligand activity.We demonstrate this methodology by using cyclic voltammetry to characterize the kinetics of a modelinterfacial reaction with aminooxy acetic acid. We also demonstrate the synthetic flexibility and utility ofthis method for biospecific interactions by installing aminooxy terminated FLAG peptides and characterizingtheir binding to soluble anti−FLAG with surface plasmon resonance spectroscopy. We further show thismethodology is compatible with microarray technology by printing rhodamine−oxyamine in various sizespots and characterizing the yield within the spots by cyclic voltammetry. We also show this methodologyis compatible with cell culture conditions and fluorescent microscopy technology for cell biological studies.Arraying RGD−oxyamine peptides on these substrates allows for bio-specific adhesion of Swiss 3T3Fibroblasts.
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