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Conformational Antibody Binding to a Native, Cell-Free Expressed GPCR in Block Copolymer Membranes

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posted on 2014-10-20, 02:48 authored by Hans-Peter M. de Hoog, Esther M. Lin JieRong, Sourabh Banerjee, Fabien M. Décaillot, Madhavan Nallani

G-protein coupled receptors (GPCRs) play a key role in physiological processes and are attractive drug targets. Their biophysical characterization is, however, highly challenging because of their innate instability outside a stabilizing membrane and the difficulty of finding a suitable expression system. We here show the cell-free expression of a GPCR, CXCR4, and its direct embedding in diblock copolymer membranes. The polymer-stabilized CXCR4 is readily immobilized onto biosensor chips for label-free binding analysis. Kinetic characterization using a conformationally sensitive antibody shows the receptor to exist in the correctly folded conformation, showing binding behaviour that is commensurate with heterologously expressed CXCR4.

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