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Ultrasensitive Immunosensor Array for TNF-α Detection in Artificial Saliva using Polymer-Coated Magnetic Microparticles onto Screen-Printed Gold Electrode

1
NANOMISENE Lab, LR16CRMN01, Centre for Research on Microelectronics and Nanotechnology of Sousse, Technopole of Sousse B.P. 334, Sahloul 4034, Sousse, Tunisia
2
University of Sousse, Higher Institute of Applied Sciences and Technology of Sousse, GREENS-ISSAT, Cité Ettafala, Ibn Khaldoun 4003, Sousse, Tunisia
3
Department of Chemistry and Industrial Chemistry, University of Pisa, via Giuseppe Moruzzi 13, 56124 Pisa, Italy
4
Université de Lyon 1, Institut des Sciences Analytiques, UMR 5280, CNRS, 5 rue de la Doua, 69100 Villeurbanne, France
5
Univ Lyon, University Claude Bernard Lyon-1, CNRS, LAGEP-UMR 5007, F69622 Lyon, France
*
Author to whom correspondence should be addressed.
Sensors 2019, 19(3), 692; https://doi.org/10.3390/s19030692
Received: 26 November 2018 / Revised: 29 January 2019 / Accepted: 1 February 2019 / Published: 8 February 2019
(This article belongs to the Section Chemical Sensors)
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Abstract

Tumor necrosis factor-α (TNF-α) is a biomarker of inflammation that occurs in patients suffering from heart failure (HF). Saliva can be sampled in a non-invasive way, and it is currently gaining importance as matrix alternative to blood in diagnostic and therapy monitoring. This work presents the development of an immunosensor array based on eight screen-printed gold electrodes to detect TNF-α in saliva samples. Two different functionalization strategies of electrodes were compared. In the first, anti-TNF-α antibodies were chemically bonded onto the electrode by functionalization with 4-carboxymethylaniline. The other functionalization procedure involved the binding of antibodies onto polymer-coated magnetic microparticles, which were then deposited onto the electrode by pulsed chronoamperometry. Finally, the chronoamperometry technique was applied to characterize the modified SPEAu. The use of a secondary antibody anti-TNF-α (Ab-TNF-α-HRP) labelled with horseradish peroxidase (HRP, 2 µg·mL−1) was investigated using tetramethylbenzidine (TMB, pH = 3.75) as electrochemical substrate containing 0.2 mM of H2O2. A sandwich-type detection strategy with a secondary antibody anti-TNF-α provided chronoamperometric analyses in 10 s for each sample. Linearity, precision, limit of detection, and selectivity of devices were investigated. Interferences were evaluated by analyzing solutions containing other cytokine produced during the acute stage of inflammation. The immunosensor showed good performance within the clinically relevant concentration range, with a precision of 8%, and a limit of detection of 0.3 pg/mL. Therefore, it may represent a promising tool for monitoring HF in a non-invasive way. View Full-Text
Keywords: magnetic microparticles; immunosensor; tumor necrosis factor-α; chronoamperometry; saliva analysis; heart failure magnetic microparticles; immunosensor; tumor necrosis factor-α; chronoamperometry; saliva analysis; heart failure
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Barhoumi, L.; Bellagambi, F.G.; Vivaldi, F.M.; Baraket, A.; Clément, Y.; Zine, N.; Ben Ali, M.; Elaissari, A.; Errachid, A. Ultrasensitive Immunosensor Array for TNF-α Detection in Artificial Saliva using Polymer-Coated Magnetic Microparticles onto Screen-Printed Gold Electrode. Sensors 2019, 19, 692.

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