A screen-printed microband glucose biosensor system for real-time monitoring of toxicity in cell culture

Xu, J.; Pemberton, Roy; Pittson, Robin; Drago, Guido A; Griffiths, John; Jackson, Simon K.; Hart, John P.

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 (Eapp of +0.4V), in a 12-well tissue culture plate system at 37°C, using a multipotentiostat. After 24h, a linear calibration plot was obtained from steady-state current responses for glucose concentrations up to 10mM (dynamic range 30mM). Within the linear region, a correlation coefficient (R2) of 0.981 was obtained between biosensor and spectrophotometric assays. Over 24h, an estimated 0.15% (89nmol) of the starting glucose concentration (24mM) 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 106cell/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 1mM paracetamol, glucose uptake was inhibited by 95% after 6h in culture; at 500μM, around 15% inhibition was observed after 16h. This microband biosensor culture system could form the basis for an in vitro toxicity testing system. © 2010 Elsevier B.V.

Citation

Xu, J., Pemberton, R., Pittson, R., Drago, G. A., Griffiths, J., Jackson, S. K., & Hart, J. P. (2011). A screen-printed microband glucose biosensor system for real-time monitoring of toxicity in cell culture. Biosensors and Bioelectronics, 26(5), 2448-2453. https://doi.org/10.1016/j.bios.2010.10.030

• Journal Article Type: Article
• Publication Date: Jan 15, 2011
• Journal: Biosensors and Bioelectronics
• Print ISSN: 0956-5663
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 26
• Issue: 5
• Pages: 2448-2453
• DOI: https://doi.org/10.1016/j.bios.2010.10.030
• Keywords: glucose biosensor, microband electrode, HepG2 cell culture, continuous monitoring, glucose metabolism
• Public URL: https://uwe-repository.worktribe.com/output/966433
• Publisher URL: http://dx.doi.org/10.1016/j.bios.2010.10.030