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Herein, a novel hybrid bilayer membrane is introduced as a platform to study the aggregation of amyloid-β1–42 (Aβ1–42) peptide on surfaces. The first layer was covalently attached to a glassy carbon electrode (GCE) via diazonium electrodeposition, which provided a highly stable template for the hybrid bilayer formation. To prepare the long-chain hybrid bilayer membrane (lcHBLM)-modified electrodes, GCE surfaces were modified with 4-dodecylbenzenediazonium (DDAN) followed by the modification with dihexadecyl phosphate (DHP) as the second layer. For the preparation of short-chain hybrid bilayer membrane (scHBLM)-modified electrodes, GCE surfaces were modified with 4-ethyldiazonium (EDAN) as the first layer and bis(2-ethylhexyl) phosphate (BEHP) was utilized as the second layer. X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS) were used to characterize the bilayer formation. Both positively charged [Ru(NH3)6]3+ and negatively charged ([Fe(CN)6]3-/4-) redox probes were used for electrochemical characterization of the modified surfaces using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). EIS results showed a decrease in charge transfer resistance (Rct) upon incubation of Aβ1–42 on the hybrid bilayer-modified surfaces. This framework provides a promising electrochemical platform for designing hybrid bilayers with various physicochemical properties to study the interaction of membrane-bound receptors and biomolecules on surfaces.

Electrografting a Hybrid Bilayer Membrane via Diazonium Chemistry for Electrochemical Impedance Spectroscopy of Amyloid-β Aggregation