title: Conformational antibody binding to a native, cell-free expressed GPCR in block copolymer membranes.
abstract: 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.
authors.affiliation: ACM Biolabs Pte Ltd, Research Techno Plaza XF-6, Singapore, Singapore.
authors.forename: Hans-Peter M
authors.initials: HP
authors.lastname: de Hoog
authors.affiliation: ACM Biolabs Pte Ltd, Research Techno Plaza XF-6, Singapore, Singapore.
authors.forename: Esther M
authors.initials: EM
authors.lastname: Lin JieRong
authors.affiliation: ACM Biolabs Pte Ltd, Research Techno Plaza XF-6, Singapore, Singapore.
authors.forename: Sourabh
authors.initials: S
authors.lastname: Banerjee
authors.affiliation: Singapore Immunology Network, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore.
authors.forename: Fabien M
authors.initials: FM
authors.lastname: Décaillot
authors.affiliation: ACM Biolabs Pte Ltd, Research Techno Plaza XF-6, Singapore, Singapore; Centre for Biomimetic Sensor Science, School of Materials Science and Engineering, Nanyang Technological University, Singapore, Singapore.
authors.forename: Madhavan
authors.initials: M
authors.lastname: Nallani
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D015374:Biosensing Techniques; D002474:Cell-Free System / Q000737:chemistry; D006801:Humans; D008567:Membranes, Artificial; D051379:Mice; D011487:Protein Conformation; D017510:Protein Folding; D019718:Receptors, CXCR4 / Q000737:chemistry*
chemical_list: D000906:Antibodies; C527276:CXCR4 protein, human; D008567:Membranes, Artificial; D019718:Receptors, CXCR4