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An electrochemical sensor array system for the direct, simultaneous in vitro monitoring of nitric oxide and superoxide production by cultured cells

Lookup NU author(s): Dr Seung Chang, Dr James Henderson, Emeritus Professor Calum McNeilORCiD

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Abstract

A new approach for an amperometric array sensor platform employing arrays of sensors in a 24-well cell culture plate format has been developed for simultaneous in vitro determination of nitric oxide (NO) and superoxide free radicals (O2-) produced by stimulated cells. The work reported focuses on the direct, real-time monitoring of extracellular production of these two analytes, as well as the effects of their interaction. The sensor platform was manufactured by a combination of sputtering gold electrodes and screen-printing carbon electrodes. The O2- sensor uses covalent immobilization of cytochrome c via a binder, DTSSP (3,3′-dithio- bis(sulphosuccinimidylpropionate) onto the surface of the Au electrodes, whereas the NO sensor system involves an NiTSPc (nickel tetrasulfonated phthalocyanine) film electrodeposited onto the surface of the carbon electrodes and subsequently covered with an external layer of Nafion®. For in vitro demonstration of the platforms as a potential drug-screening system, A172 glioblastoma cells were cultured and transferred into the 24-well arrays. Simultaneous and direct monitoring of NO and O2- production as a response to chemicals of biomedical relevance was carried out. The results obtained demonstrated that it would be possible to envisage a drug screening platform for compounds designed to be inhibitors of nitric oxide synthase or to have an inhibitory effect on superoxide free radical production. By suitable modification of the electrodes employed it would also be possible to extend the platform to measure alternative species. © 2005 Elsevier B.V. All rights reserved.


Publication metadata

Author(s): Chang S-C, Pereira-Rodrigues N, Henderson JR, Cole A, Bedioui F, McNeil CJ

Publication type: Article

Publication status: Published

Journal: Biosensors and Bioelectronics

Year: 2005

Volume: 21

Issue: 6

Pages: 917-922

Print publication date: 15/12/2005

ISSN (print): 0956-5663

ISSN (electronic): 1873-4235

Publisher: Elsevier

URL: http://dx.doi.org/10.1016/j.bios.2005.02.015

DOI: 10.1016/j.bios.2005.02.015

PubMed id: 16257661


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