Microfabricated glucose biosensor for culture welloperation

Pemberton, R. M.; Tuffin, R.; Sage, I.; Griffiths, J.; Xu, J.; Luxton, R.; Hart, J. P.; Pemberton, Roy; Tuffin, Rachel; Sage, Ian; Griffiths, John; Cox, T.; Xu, Jinsheng; Drago, G. A.; Biddle, N.; Luxton, Richard W.; Hart, John P.; Pittson, R.; Johnson, G.; Jackson, S. K.; Kenna, G.

Abstract

A water-based carbon screen-printing ink formulation, containing the redox mediator cobalt phthalocyanine (CoPC) and the enzyme glucose oxidase (GOx), was investigated for its suitability to fabricate glucose microbiosensors in a 96-well microplate format: (1)the biosensor ink was dip-coated onto a platinum (Pt) wire electrode, leading to satisfactory amperometric performance; (2)the ink was deposited onto the surface of a series of Pt microelectrodes (10-500 μm diameter) fabricated on a silicon substrate using MEMS (microelectromechanical systems) microfabrication techniques: capillary deposition proved to be successful; a Pt microdisc electrode of ≥100 μm was required for optimum biosensor performance; (3)MEMS processing was used to fabricate suitably sized metal (Pt) tracks and pads onto a silicon 96 well format base chip, and the glucose biosensor ink was screen-printed onto these pads to create glucose microbiosensors. When formed into microwells, using a 340 μl volume of buffer, the microbiosensors produced steady-state amperometric responses which showed linearity up to 5. mM glucose (CV=6% for n=5 biosensors). When coated, using an optimised protocol, with collagen in order to aid cell adhesion, the biosensors continued to show satisfactory performance in culture medium (linear range to 2. mM, dynamic range to 7. mM, CV=5.7% for n=4 biosensors). Finally, the operation of these collagen-coated microbiosensors, in 5-well 96-well format microwells, was tested using a 5-channel multipotentiostat. A relationship between amperometric response due to glucose, and cell number in the microwells, was observed. These results indicate that microphotolithography and screen-printing techniques can be combined successfully to produce microbiosensors capable of monitoring glucose metabolism in 96 well format cell cultures. The potential application areas for these microbiosensors are discussed. © 2012 Elsevier B.V.

• Journal Article Type: Article
• Publication Date: Apr 5, 2013
• Journal: Biosensors and Bioelectronics
• Print ISSN: 0956-5663
• Electronic ISSN: 1873-4235
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 42
• Issue: 1
• Pages: 668-677
• DOI: https://doi.org/10.1016/j.bios.2012.11.032
• Keywords: glucose microbiosensor, water-based screen-printing ink, ; 96-Well format, cell culture
• Public URL: https://uwe-repository.worktribe.com/output/932797
• Publisher URL: http://dx.doi.org/10.1016/j.bios.2012.11.032