title: A living cell-based biosensor utilizing G-protein coupled receptors: principles and detection methods.
abstract: This study explores the feasibility of using a bullfrog fibroblast cell line (FT cells) expressing G protein coupled receptors (GPCRs) as the basis for a living cell-based biosensor. We have fabricated gold microelectrode arrays on a silicon dioxide substrate that supports long term, robust growth of the cells at room temperature and under ambient atmospheric conditions. Activation of an endogenous GPCR to ATP was monitored with an optical method that detects rises in intracellular calcium and with an electrochemical method that monitors the increased secretion of pre-loaded norepinephrine on a MEMS device. FT cells were also transfected to express reporter genes driven by several different promoters, raising the possibility that they could be modified genetically to express novel GPCRs as well. The ability to harness GPCRs for BioMEMS applications by using cells that are easy to grow on MEMS devices and to modify genetically opens the way for a new generation of devices based on these naturally selective and highly sensitive chemoreceptors.
authors.affiliation: College of Nanoscale Science and Engineering, University at Albany, State University of New York, 255 Fuller Road, Albany, NY 12203, United States.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Feng
authors.forename: James
authors.initials: J
authors.lastname: Castracane
authors.forename: Natalya
authors.initials: N
authors.lastname: Tokranova
authors.forename: Alison
authors.initials: A
authors.lastname: Gracias
authors.forename: Gregory
authors.initials: G
authors.lastname: Lnenicka
authors.forename: Ben G
authors.initials: BG
authors.lastname: Szaro
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D002118:Calcium / Q000378:metabolism; D004563:Electrochemistry; D005347:Fibroblasts / Q000378:metabolism; D046228:Microarray Analysis; D011892:Rana catesbeiana; D043562:Receptors, G-Protein-Coupled / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D018048:Receptors, Purinergic P2 / Q000187:drug effects
chemical_list: D043562:Receptors, G-Protein-Coupled; D018048:Receptors, Purinergic P2; D000255:Adenosine Triphosphate; D002118:Calcium