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Label-free quantification of membrane-ligand interactions using backscattering interferometry
Label-free quantification of membrane-ligand interactions using backscattering interferometry
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Label-free quantification of membrane-ligand interactions using backscattering interferometry
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Label-free quantification of membrane-ligand interactions using backscattering interferometry
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Label-free quantification of membrane-ligand interactions using backscattering interferometry
Label-free quantification of membrane-ligand interactions using backscattering interferometry
Journal Article

Label-free quantification of membrane-ligand interactions using backscattering interferometry

2011
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Overview
Methods to measure affinities of membrane proteins and soluble ligands are cumbersome and often rely on truncations or other modifications of the membrane protein or ligand. Baksh et al . show that backscattering interferometry is a sensitive and accurate technology for the label-free quantification of ligand–membrane receptor interactions. Although membrane proteins are ubiquitous within all living organisms and represent the majority of drug targets, a general method for direct, label-free measurement of ligand binding to native membranes has not been reported. Here we show that backscattering interferometry (BSI) can accurately quantify ligand-receptor binding affinities in a variety of membrane environments. By detecting minute changes in the refractive index of a solution, BSI allows binding interactions of proteins with their ligands to be measured at picomolar concentrations. Equilibrium binding constants in the micromolar to picomolar range were obtained for small- and large-molecule interactions in both synthetic and cell-derived membranes without the use of labels or supporting substrates. The simple and low-cost hardware, high sensitivity and label-free nature of BSI should make it readily applicable to the study of many membrane-associated proteins of biochemical and pharmacological interest.