2026-06-08T04:19:48Zhttps://docta.ucm.es/rest/oai/request

oai:docta.ucm.es:20.500.14352/918142025-03-18T12:11:47Zcom_20.500.14352_14col_20.500.14352_15

00925njm 22002777a 4500 dc Povedano Muñumel, Eloy author Gamella Carballo, María author Torrente Rodríguez, Rebeca Magnolia author Montero-Calle, Ana author Pedrero Muñoz, María author Solís-Fernández, Guillermo author Navarro Villoslada, Fernando author Barderas Manchado, Rodrigo author Campuzano Ruiz, Susana author Pingarrón Carrazón, José Manuel author 2021 This work describes the preparation of an immunoplatform for the sensitive and selective determination of N6-methyladenosine (m6A). The simple and fast protocol involves for the first time the use of micromagnetic immunoconjugates to establish a direct competitive assay between the m6A target and a biotinylated RNA oligomer bearing a single m6A enzymatically labelled with a commercial conjugate of streptavidin-peroxidase (Strep-HRP) as tracer. The cathodic current change measured in the presence of H2O2/hydroquinone (HQ) at screen-printed carbon electrodes (SPCEs) upon surface capturing the magnetic bioconjugates is inversely proportional to the m6A target concentration. After evaluating the effect of key variables, the analytical characteristics were established for the determination of three different targets: the N6-methyladenosine-5′ -triphosphate (m6ATP) ribonucleotide, a short synthetic RNA oligomer bearing a single m6A and the positive control provided in a commercial colorimetric kit for m6A-RNA quantification. The obtained results show that this immunoplatform is competitive with other methods reported to date, achieving an improved sensitivity (limit of detection of 0.9 pM for the short synthetic oligomer) using a much simpler and faster protocol (~1 h) and disposable electrodes for the transduction. Furthermore, the applicability for discriminating the metastatic potential of cancer cells by directly analyzing a small amount of raw total RNA without enriching or fragmenting was also preliminary assessed. Povedano, Eloy, et al. «Magnetic Microbeads-Based Amperometric Immunoplatform for the Rapid and Sensitive Detection of N6-Methyladenosine to Assist in Metastatic Cancer Cells Discrimination». Biosensors and Bioelectronics, vol. 171, enero de 2021, p. 112708. https://doi.org/10.1016/j.bios.2020.112708. 0956-5663 10.1016/j.bios.2020.112708 https://hdl.handle.net/20.500.14352/91814 https://doi.org/10.1016/j.bios.2020.112708 Magnetic microbeads-based amperometric immunoplatform for the rapid and sensitive detection of N6-methyladenosine to assist in metastatic cancer cells discrimination