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

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...

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Bibliographic Details
Authors: Pedrero Muñoz, María, Manuel De Villena Rueda, Francisco Javier, Muñoz San Martín, Cristina, Campuzano Ruiz, Susana, Garranzo Asensio, María, Barderas Manchado, Rodrigo, Pingarrón Carrazón, José Manuel
Format: article
Publication Date:2016
Country:España
Institution:Universidad Complutense de Madrid (UCM)
Repository:Docta Complutense
Language:English
OAI Identifier:oai:docta.ucm.es:20.500.14352/19176
Online Access:https://hdl.handle.net/20.500.14352/19176
Access Level:Open access
Keyword:human p53
magnetic microcarriers
screen-printed electrodes
amperometric immunosensor
cell lysates
Bioquímica (Química)
Química analítica (Química)
2301 Química Analítica
Description
Summary: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.