Piezoelectric Biosensor Based on Olfactory Receptor Expressed in a Heterologous Cell System for Drug Discovery

摘要

Nowadays,there is an urgent need to find novel tools and approaches to improve the overall efficiency of drug discovery.In the present study,we developed a quartz crystal microbalance (QCM)-based biosensor in which olfactory receptor expressed in a heterologous cell system was used as sensitive elements.An olfactory receptor protein of C.elegances,ODR-10,which was serve as a model of G-proteincoupled receptors (GPCRs),was expressed on the plasma membrane of human embryonic kidney (HEK)-293 cells.For targeting the ODR-10 on the surface of cell membrane,the rho-tag import sequence was inserted into the N-terminus of the ODR-10.The membrane expression of ODR-10 was shown by the enhanced green fluorescent protein (EGFP) which sequences was inserted into the C-terminus of the ODR-10.After that,the membrane fraction of HEK-293 cells containing ODR-10 was extracted and then coated on the electrode surface of QCM.The interactions between odorant molecules and olfactory receptors were monitored by QCM.The results indicate that ODR-10 was functionally expressed on the plasma membrane of HEK-293 cells and it can respond to diacetyl with high specificity,which is a natural ligand for ODR-10.All the results suggest one conclusion:this novel QCM-olfactory receptor hybrid biosensor can be used to detect the interactions between GPCRs and their ligands,and suggest it could be a potential tool for drug discovery and even for drug evaluation.