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-protein-coupled 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.
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- Piezoelectric Biosensor Based on Olfactory Receptor Expressed in a Heterologous Cell System for Drug Discovery
- Springer Berlin Heidelberg