title: Disposable Amperometric Immunosensor for the Determination of Human P53 Protein in Cell Lysates Using Magnetic Micro-Carriers.
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.
authors.affiliation: Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. mpedrero@quim.ucm.es.
authors.forename: María
authors.initials: M
authors.lastname: Pedrero
authors.affiliation: Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. villena@quim.ucm.es.
authors.forename: F Javier Manuel
authors.initials: FJM
authors.lastname: de Villena
authors.affiliation: Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. cmunoz04@ucm.es.
authors.forename: Cristina
authors.initials: C
authors.lastname: Muñoz-San Martín
authors.affiliation: Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. susanacr@quim.ucm.es.
authors.forename: Susana
authors.initials: S
authors.lastname: Campuzano
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. mgarranzo@hotmail.com.
authors.forename: María
authors.initials: M
authors.lastname: Garranzo-Asensio
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. rbarderas@quim.ucm.es.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Barderas
authors.affiliation: Departamento de Química Analítica, Facultad de CC. Químicas, Universidad Complutense de Madrid, E-28040 Madrid, Spain. pingarro@quim.ucm.es.
authors.forename: José M
authors.initials: JM
authors.lastname: Pingarrón
mesh_terms: D015374:Biosensing Techniques; D002460:Cell Line; D004566:Electrodes; D006801:Humans; D008863:Microspheres; D012680:Sensitivity and Specificity; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D016159:Tumor Suppressor Protein p53
keywords: amperometric immunosensor; cell lysates; human p53; magnetic microcarriers; screen-printed electrodes