Abstract

Supported lipid bilayers (SLBs) have been used extensively as an effective model of biological membranes, in the context of in vitro biophysics research, and the membranes of liposomes, in the context of the development of nanoscale drug delivery devices. Despite numerous surface-sensitive techniques having been applied to their study, the comprehensive optical characterization of SLBs using surface plasmon resonance (SPR) has not been conducted. In this study, Fresnel multilayer analysis is utilized to effectively calculate layer parameters (thickness and refractive indices) with the aid of dual-wavelength and dispersion coefficient analysis, in which the linear change in the refractive index as a function of wavelength is assumed. Using complementary information from impedance-based quartz crystal microbalance experiments, biophysical properties, for example, area-per-lipid-molecule and the quantity of lipid-associated water molecules, are calculated for different lipid types and mixtures, one of which is representative of a raft-forming lipid mixture. It is proposed that the hydration layer beneath the bilayer is, in fact, an integral part of the measured optical signal. Also, the traditional Jung model analysis and the ratio of SPR responses are investigated in terms of assessing the structure of the lipid layer that is formed.

Highlights

  • Biomimetic membrane models are the leading platforms to complement in vitro cell-screening assays in the analysis of biochemical and physical interactions involving biomembranes.[1,2] The design and study of these platforms is important with respect to advancements in cell biology and from a pharmaceutical perspective; individualized drug therapies and selective targeting of membrane proteins require information regarding the complex catalytic biochemical processes performed by membrane proteins with a resolution far beyond the diffraction limit of visual optics

  • The two methods (3 and 4) used in the analysis yield very similar results: refractive index values at 670 nm wavelength for different lipid bilayer compositions are in the range of 1.4776−1.4889, using method (3) and 1.4774−1.4855, using method (4)

  • We have provided advanced methodology for thin-layer characterization utilizing Fresnellayer analysis, enabling the accurate determination of the thickness, refractive indices, and linear dispersion coefficient of the layer

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Summary

Introduction

Biomimetic membrane models are the leading platforms to complement in vitro cell-screening assays in the analysis of biochemical and physical interactions involving biomembranes.[1,2] The design and study of these platforms is important with respect to advancements in cell biology and from a pharmaceutical perspective; individualized drug therapies and selective targeting of membrane proteins require information regarding the complex catalytic biochemical processes performed by membrane proteins with a resolution far beyond the diffraction limit of visual optics. Intrinsic autofluorescence of the compound libraries themselves can result in false positives through interference effects. These factors increase the need for label-free alternatives resilient to autofluorescence.[3] membrane models are a key tool for the in vitro investigation of the surface properties of nanoscale drug delivery devices, for example, drug delivery liposomes.[4]

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