Application of screen-printed microband biosensors incorporated with cells to monitor metabolic effects of potential environmental toxins

• Published in: Mikrochimica acta (1966) Vol. 170; no. 3-4; pp. 321 - 330
• Main Authors: Pemberton, Roy M, Rawson, Frankie J, Xu, Jinsheng, Pittson, Robin, Drago, Guido A, Griffiths, John, Jackson, Simon K, Hart, John P
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Vienna Vienna : Springer Vienna 01-09-2010; Springer Vienna; Springer
• Subjects: Analytical Chemistry; Applied sciences; Biological and medical sciences; Biosensors; Biotechnology; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Continuous monitoring; Cyanides; Electrochemical methods; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; General, instrumentation; Global environmental pollution; HepG2 cell culture; Lactates; Medical equipment; Methods. Procedures. Technologies; Microband electrode biosensor; Microengineering; Nanochemistry; Nanotechnology; Original Paper; Oxidases; Phosphates; Phthalocyanins; Physiological apparatus; Pollution; toxicity; Various methods and equipments; Microband electrode biosensor; Continuous monitoring; Toxicity; HepG2 cell culture; Water; Phosphates; Cell culture; Glucose oxidase; High density; Ink; Time; Cobalt; Cyclic voltammetry; Phthalocyanine; Reduction; Dynamics; Platinum; Production; Amperometry; Microelectrode; Diffusion; Release; Monitoring; Culture medium; Enzyme; Use; Buffer solution; Carbon; Steady state; Lactates; Toxin; Sensitivity; Biosensor; Environment; Oxidoreductases; Hexacyanoferrates II; Application
• ISSN: 0026-3672; 1436-5073
• DOI: 10.1007/s00604-010-0326-0

Microband biosensors were fabricated from a screen-printed water-based carbon ink containing cobalt phthalocyanine redox mediator and glucose oxidase or lactate oxidase enzyme. The microbiosensors were characterised for their ability to monitor ferrocyanide and H₂O₂ in phosphate buffer solution: sigmoidal cyclic voltammograms, high current density values and steady-state amperometric responses confirmed the existence of radial-diffusion-limiting microelectrode behaviour. The lactate microband biosensors were then used, in conjunction with a screen-printed Ag/AgCl reference and platinum counter electrode, to monitor lactate levels in culture medium, with a linear range of 0.5-5 mM, sensitivity of 20 nA.mM⁻¹, and dynamic range up to >9 mM. The lactate microband biosensors could operate continuously in culture medium over extended times (up to 24 h) at 37 °C. These biosensors were then applied to detect changes in lactate release from cultured cells in response to toxic challenge: m-dinitrobenzene (500 μM) caused a reduction in lactate production by high-passage number HepG2 single cells; D-galactosamine (20 mM) induced release of lactate by HepG2 spheroid cultures. This novel use of microband biosensors in cell culture has the potential for further application in toxicity monitoring, in both environmental and pharmaceutical areas.