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oai:docta.ucm.es:20.500.14352/1136472025-03-18T15:00:22Zcom_20.500.14352_14col_20.500.14352_15

Magnetic beads-based electrochemical immunosensing of HIF-1α, a biomarker of tumoral hipoxia Muñoz San Martín, Cristina Gamella Carballo, María Pedrero Muñoz, María Montero Calle, Ana Barderas Manchado, Rodrigo Campuzano Ruiz, Susana Pingarrón Carrazón, José Manuel 543 HIF-1α MBs-based immunoassay Amperometric detection Cancer cells Tumoral hypoxia Saliva Ciencias 23 Química Hypoxia-inducible factor-1 alpha (HIF-1α) is a transcription factor involved in tumor growth and metastasis by regulating genes involved in response to hypoxia. This work reports the first magnetic beads (MBs)-based electrochemical immunoassay for the determination of HIF-1α. The design involves a sandwich immunoassay using MBs and amperometric detection at disposable screen-printed carbon electrodes (SPCEs). Magnetic immunoconjugates of carboxylic acid-microsized magnetic particles (HOOC-MBs) modified with a specific antibody were used to selectively capture the target protein which was sandwiched with a biotinylated secondary antibody further conjugated with a streptavidin peroxidase conjugate (Strep-HRP). The great analytical performance of the immunoassay in terms of selectivity and low limit of detection (76 pg mL−1) for HIF-1α allows the determination of the endogenous amount of this biomarker in 0.5 μg raw cancer cells lysates cultured in normoxia or induced-hypoxia with CoCl2. In addition, the developed immunosensing approach was employed to analyze HIF-1α in saliva samples with results that agree with those provided by an ELISA kit involving the same immunoreagents. Ministerio de Economía y Competitividad Comunidad de Madrid Universidad Complutense de Madrid Ministerio de Educación, Cultura y Deporte Depto. de Química Analítica Fac. de Ciencias Químicas TRUE pub 2025-01-10T09:26:17Z 2025-01-10T09:26:17Z 2020-03-20 journal article VoR https://hdl.handle.net/20.500.14352/113647 XXXX-XXXX 10.1016/j.snb.2019.127623 eng info:eu-repo/grantAgreement/MINECO//CTQ2015-64402-C2-1-R/ES/NUEVAS PLATAFORMAS DE MULTIPLEXADO ELECTROANALITICAS PARA LA DETECCION Y PRONOSTICO DE ENFERMEDADES NEOPLASICAS MEDIANTE BIOPSIAS LIQUIDAS/ Grant S2018/NMT-4349 PI17CIII/00045 Cristina Muñoz-San Martín, Maria Gamella, María Pedrero, Ana Montero-Calle, Rodrigo Barderas, Susana Campuzano, José M. Pingarrón, Magnetic beads-based electrochemical immunosensing of HIF-1α, a biomarker of tumoral hypoxia, Sensors and Actuators B: Chemical, Volume 307, 2020, 127623, ISSN 0925-4005, https://doi.org/10.1016/j.snb.2019.127623. (https://www.sciencedirect.com/science/article/pii/S0925400519318222) Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ restricted access application/pdf Elsevier