title: A novel surface acoustic wave-based biosensor for highly sensitive functional assays of olfactory receptors.
abstract: Olfactory receptors, which are responsible for sensing odor molecules, form the largest G protein-coupled receptor (GPCR) family in mammalian animals. These proteins play an important role in the detection of chemical signals and signal transduction to the brain. Currently, only a limited number of olfactory receptors have been characterized, which is mainly due to the lack of sensitive and efficient tools for performing functional assays of these receptors. This paper describes a novel surface acoustic wave (SAW)-based biosensor for highly sensitive functional assays of olfactory receptors. An olfactory receptor of Caenorhabditis elegans, ODR-10, was expressed on the plasma membrane of human breast cancer MCF-7 cells, which was used as a model system for this study. For specific odorant response assays, the membrane fraction of MCF-7 cells containing ODR-10 was extracted and integrated with our SAW sensors. The response of ODR-10 to various odorants was monitored by recording the resonance frequency shifts of SAWs applied to the sensor. Our results show that heterologously expressed ODR-10 receptors can specifically respond to diacetyl, its natural ligand. Dose-dependent responses were obtained by performing measurements using various concentrations of diacetyl. The sensitivity of this biosensor is 2kHz/ng and can detect concentrations as low as 10(-10)mM, which is 10× lower than what has previously been reported. This biosensor can be used to characterize odorant response profiles of olfactory receptors and provide information rich data for functional assays of olfactory receptors. In addition to providing a greater understanding of the biological mechanisms of GPCRs, such data holds great potential in many other fields such as food industry, biomedicine, and environmental protection.
authors.affiliation: Biosensor National Special Laboratory, Key Laboratory for Biomedical Engineering of Ministry of Education, Department of Biomedical Engineering, Zhejiang University, Hangzhou 310027, China. cswu@zju.edu.cn
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Wu
authors.forename: Liping
authors.initials: L
authors.lastname: Du
authors.forename: Di
authors.initials: D
authors.lastname: Wang
authors.forename: Le
authors.initials: L
authors.lastname: Wang
authors.forename: Luhang
authors.initials: L
authors.lastname: Zhao
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
mesh_terms: D000162:Acoustics; D000818:Animals; D001681:Biological Assay; D015374:Biosensing Techniques; D029742:Caenorhabditis elegans Proteins / Q000096:biosynthesis / Q000235:genetics; D045744:Cell Line, Tumor; D006801:Humans; D009812:Odorants / Q000032:analysis*; D018035:Receptors, Odorant / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D012680:Sensitivity and Specificity
chemical_list: D029742:Caenorhabditis elegans Proteins; C109039:ODR-10 protein, C elegans; D018035:Receptors, Odorant