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oai:docta.ucm.es:20.500.14352/191762025-03-06T16:36:50Zcom_20.500.14352_14col_20.500.14352_15

Disposable Amperometric Immunosensor for the Determination of Human P53 Protein in Cell Lysates Using Magnetic Micro-Carriers 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 human p53 magnetic microcarriers screen-printed electrodes amperometric immunosensor cell lysates Bioquímica (Química) Química analítica (Química) 2301 Química Analítica 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. Ministerio de Economía y Competitividad (MINECO) Comunidad de Madrid Depto. de Bioquímica y Biología Molecular Depto. de Química Analítica Fac. de Ciencias Químicas TRUE pub 2023-06-18T00:02:05Z 2023-06-18T00:02:05Z 2016-11-21 journal article https://hdl.handle.net/20.500.14352/19176 2079-6374 10.3390/bios6040056 eng CTQ2015-64402-C2-1-R NANOAVANSENS-CM (S2013/MT–3029) Atribución 3.0 España https://creativecommons.org/licenses/by/3.0/es/ open access application/pdf MDPI