RT Journal Article
T1 A novel zinc finger protein–based amperometric biosensor for miRNA determination
A1 Montoya Miñano, Juan José
A1 Moranova, Ludmila
A1 Bartosik, Martin
A1 Povedano Muñumel, Eloy
A1 Ruiz Valdepeñas Montiel, Víctor
A1 Gamella Carballo, María
A1 Serafín González-Carrato, Verónica
A1 Pedrero Muñoz, María
A1 Yáñez-Sedeño Orive, Paloma
A1 Campuzano Ruiz, Susana
A1 Pingarrón Carrazón, José Manuel
AB This paper reports a simple electrochemical strategy for the determination of microRNAs (miRNAs) using a commercial His-Tag-Zinc finger protein (His-Tag-ZFP) that binds preferably (but non-sequence specifically) RNA hybrids over ssRNAs, ssDNAs, and dsDNAs. The strategy involves the use of magnetic beads (His-Tag-Isolation-MBs) as solid support to capture the conjugate formed in homogenous solution between His-Tag-ZFP and the dsRNA homohybrid formed between the target miRNA (miR-21 selected as a model) and a biotinylated synthetic complementary RNA detector probe (b-RNA-Dp) further conjugated with a streptavidin–horseradish peroxidase (Strep–HRP) conjugate. The electrochemical detection is carried out by amperometry at disposable screen-printed carbon electrodes (SPCEs) (− 0.20 V vs Ag pseudo-reference electrode) upon magnetic capture of the resultant magnetic bioconjugates and H2O2 addition in the presence of hydroquinone (HQ). The as-prepared biosensor exhibits a dynamic concentration range from 3.0 to 100 nM and a detection limit (LOD) of 0.91 nM for miR-21 in just ~ 2 h. An acceptable discrimination was achieved between the target miRNA and other non-target nucleic acids (ssDNA, dsDNA, ssRNA, DNA–RNA, miR-122, miR-205, and single central- or terminal-base mismatched sequences). The biosensor was applied to the analysis of miR-21 from total RNA (RNAt) extracted from epithelial non-tumorigenic and adenocarcinoma breast cells without target amplification, pre-concentration, or reverse transcription steps. The versatility of the methodology due to the ZFP’s non-sequence-specific binding behavior makes it easily extendable to determine any target RNA only by modifying the biotinylated detector probe.
PB Springer Nature Link
SN 1618-2642
YR 2019
FD 2019-11-19
LK https://hdl.handle.net/20.500.14352/115015
UL https://hdl.handle.net/20.500.14352/115015
LA eng
NO Spanish Ministerio de Economía y Competitividad
NO Ministerio de Ciencia, Innovación y Universidades
NO TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid
NO Czech Science Foundation
NO Universidad Complutense de Madrid
NO Postdoctoral contract type Art 83
DS Docta Complutense
RD 27 feb 2026