title: A screen-printed microband glucose biosensor system for real-time monitoring of toxicity in cell culture.
abstract: Microband biosensors, screen-printed from a water-based carbon ink containing cobalt phthalocyanine redox mediator and glucose oxidase (GOD) enzyme, were used to monitor glucose levels continuously in buffer and culture medium. Five biosensors were operated amperometrically (E(app) of +0.4V), in a 12-well tissue culture plate system at 37°C, using a multipotentiostat. After 24 h, a linear calibration plot was obtained from steady-state current responses for glucose concentrations up to 10 mM (dynamic range 30 mM). Within the linear region, a correlation coefficient (R(2)) of 0.981 was obtained between biosensor and spectrophotometric assays. Over 24 h, an estimated 0.15% (89 nmol) of the starting glucose concentration (24 mM) was consumed by the microbiosensor. The sensitivity of the biosensor response in full culture medium was stable between pHs 7.3 and 8.4. Amperometric responses for HepG2 monolayer cultures decreased with time in inverse proportionality to cell number (for 0 to 10(6) cell/ml), as glucose was being metabolised. HepG2 3D cultures (spheroids) were also shown to metabolise glucose, at a rate which was independent of spheroid age (between 6 and 15 days). Spheroids were used to assay the effect of a typical hepatotoxin, paracetamol. At 1 mM paracetamol, glucose uptake was inhibited by 95% after 6 h in culture; at 500 μM, around 15% inhibition was observed after 16 h. This microband biosensor culture system could form the basis for an in vitro toxicity testing system.
authors.affiliation: Centre for Analytical, Materials and Sensors Science, Faculty of Health and Life Sciences, University of the West of England, Bristol BS16 1QY, UK.
authors.forename: R M
authors.initials: RM
authors.lastname: Pemberton
authors.forename: J
authors.initials: J
authors.lastname: Xu
authors.forename: R
authors.initials: R
authors.lastname: Pittson
authors.forename: G A
authors.initials: GA
authors.lastname: Drago
authors.forename: J
authors.initials: J
authors.lastname: Griffiths
authors.forename: S K
authors.initials: SK
authors.lastname: Jackson
authors.forename: J P
authors.initials: JP
authors.lastname: Hart
mesh_terms: D000082:Acetaminophen / Q000633:toxicity*; D001681:Biological Assay / Q000295:instrumentation*; D015374:Biosensing Techniques / Q000295:instrumentation*; D002470:Cell Survival / Q000187:drug effects*; D003199:Computer Systems; D003217:Conductometry / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005947:Glucose / Q000032:analysis*; D056945:Hep G2 Cells; D006801:Humans
chemical_list: D000082:Acetaminophen; D005947:Glucose