Conformational antibody binding to a native, cell-free expressed GPCR in block copolymer membranes

• Main Authors: De Hoog, Hans-Peter M., Lin JieRong, Esther M., Banerjee, Sourabh, Décaillot, Fabien M., Nallani, Madhavan
• Other Authors: Karnik, Sadashiva
• Format: Article
• Language: English
• Published: 2014
• Subjects: DRNTU::Engineering::Materials::Biomaterials
• Online Access: https://hdl.handle.net/10356/101602; http://hdl.handle.net/10220/24190
• Institution: Nanyang Technological University

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.