Tracking globally 5-methylcytosine and its oxidized derivatives in colorectal cancer epigenome using bioelectroanalytical technologies

This work presents the first electroanalytical bioplatforms to track individually or simultaneously at a global level all four methylation marks involved in the DNA methylation–demethylation cycle: 5-methylcytosine (5mC) and their sequential oxidative derivatives (5-hydroxymethyl-(5hmC), 5-formyl-(5...

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Detalles Bibliográficos
Autores: Povedano Muñumel, Eloy, Pérez Ginés, Víctor, Torrente Rodríguez, Rebeca Magnolia, Montero Calle, Ana, Peláez García, Alberto, Feliú, Jaime, Pedrero Muñoz, María, Pingarrón Carrazón, José Manuel, Barderas Manchado, Rodrigo, Campuzano Ruiz, Susana
Tipo de recurso: artículo
Fecha de publicación:2025
País:España
Institución:Universidad Complutense de Madrid (UCM)
Repositorio:Docta Complutense
Idioma:inglés
OAI Identifier:oai:docta.ucm.es:20.500.14352/123527
Acceso en línea:https://hdl.handle.net/20.500.14352/123527
Access Level:acceso abierto
Palabra clave:543
5-methylcytosine
5-hydroxymethylcytosine
5-formylcytosine
5-carboxylcytosine
bioelectroanalytical technologies
Ciencias
23 Química
Descripción
Sumario:This work presents the first electroanalytical bioplatforms to track individually or simultaneously at a global level all four methylation marks involved in the DNA methylation–demethylation cycle: 5-methylcytosine (5mC) and their sequential oxidative derivatives (5-hydroxymethyl-(5hmC), 5-formyl-(5fC), and 5-carboxyl-(5caC) cytosines). The bioplatforms employed direct competitive immunoassay formats implemented on the surface of magnetic microparticles (MBs) and involved capture antibodies specific to each epimark as well as synthetic biotinylated DNA oligomers with a single epimark that were enzymatically marked with horseradish peroxidase (HRP) to perform an amperometric readout on disposable platforms for single or multiplexed detection. These new electroanalytical biotechnologies, groundbreaking from analytical and clinical perspectives, achieved attractive operational characteristics, reaching detection limits at pM levels for synthetic single epimark-bearing DNA oligomers. The developed methodology was applied to track globally all four target epimarks in a fast, simple, sensitive, and selective way while their correlation in genomic DNA extracted from paired healthy and tumor tissues of patients with colorectal cancer (CRC) was established for the first time.