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Biosensors 2016, 6(4), 56; doi:10.3390/bios6040056

Disposable Amperometric Immunosensor for the Determination of Human P53 Protein in Cell Lysates Using Magnetic Micro-Carriers

1
Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain
2
Departamento de Bioquímica y Biología Molecular, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain
*
Author to whom correspondence should be addressed.
Academic Editor: Luigi Campanella
Received: 27 September 2016 / Revised: 4 November 2016 / Accepted: 14 November 2016 / Published: 21 November 2016
(This article belongs to the Special Issue Next-Generation Immunosensors)
View Full-Text   |   Download PDF [1235 KB, uploaded 21 November 2016]   |  

Abstract

An amperometric magnetoimmunosensor for the determination of human p53 protein is described in this work using a sandwich configuration involving the covalent immobilization of a specific capture antibody onto activated carboxylic-modified magnetic beads (HOOC-MBs) and incubation of the modified MBs with a mixture of the target protein and horseradish peroxidase-labeled antibody (HRP-anti-p53). The resulting modified MBs are captured by a magnet placed under the surface of a disposable carbon screen-printed electrode (SPCE) and the amperometric responses are measured at −0.20 V (vs. an Ag pseudo-reference electrode), upon addition of hydroquinone (HQ) as a redox mediator and H2O2 as the enzyme substrate. The magnetoimmunosensing platform was successfully applied for the detection of p53 protein in different cell lysates without any matrix effect after a simple sample dilution. The results correlated accurately with those provided by a commercial ELISA kit, thus confirming the immunosensor as an attractive alternative for rapid and simple determination of this protein using portable and affordable instrumentation. View Full-Text
Keywords: human p53; magnetic microcarriers; screen-printed electrodes; amperometric immunosensor; cell lysates human p53; magnetic microcarriers; screen-printed electrodes; amperometric immunosensor; cell lysates
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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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MDPI and ACS Style

Pedrero, M.; Manuel de Villena, F.J.; Muñoz-San Martín, C.; Campuzano, S.; Garranzo-Asensio, M.; Barderas, R.; Pingarrón, J.M. Disposable Amperometric Immunosensor for the Determination of Human P53 Protein in Cell Lysates Using Magnetic Micro-Carriers. Biosensors 2016, 6, 56.

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