Person:
Pedrero Muñoz, María

First Name

María

Last Name

Pedrero Muñoz

URI

https://hdl.handle.net/20.500.14352/76134

Affiliation

Universidad Complutense de Madrid

Faculty / Institute

Ciencias Químicas

Department

Química Analítica

Area

Química Analítica

Identifiers

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Now showing 1 - 10 of 30

Amperometric Immunosensing Scaffolds for Rapid, Simple, Non-Invasive and Accurate Determination of Protein Biomarkers of Well-Accepted and Emerging Clinical Importance
(Proceedings, 2017) Pedrero Muñoz, María; Muñoz San Martín, Cristina; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Vargas Orgaz, Eva; Manuel de Villena Rueda, Francisco Javier; Barderas Manchado, Rodrigo; Campuzano Ruiz, Susana; Pingarrón Carrazón, José Manuel
Dextran-coated nanoparticles as immunosensing platforms: Consideration of polyaldehyde density, nanoparticle size and functionality
(Talanta, 2022) Gao, Shipeng; Torrente Rodríguez, Rebeca M.; Pedrero Muñoz, María; Pingarrón Carrazón, José Manuel; Campuzano Ruiz, Susana; Rocha Martin, Javier; Guisán, José M.
Magnetic nanoparticles (MNPs) can be used as antibody carriers in a wide range of immunosensing applications. The conjugation chemistry for preparing antibody-MNP bionanohybrids should assure the nanoparticle’s colloidal dispersity, directional conformation and high biofunctionality retention of attached antibodies. In this work, peroxidase (HRP) was selected as model target analyte, and stable antibody-MNP conjugates were prepared using polyaldehyde-dextrans as multivalent linkers, also to prevent nanoparticles agglomeration and steric shielding of non-specific proteins. Under the manipulation of the oxidation variables, MNP-conjugated antibody showed the highest Fab accessibility, of 1.32 μmol analyte per μmol antibody, corresponding to 139 μmol aldehyde per gram of nanocarrier (5 mM NaIO4, 4 h). Demonstrating anti-interference advantage up to 10% serum, colorimetric immunoassay gave a detection limit (LOD) of 300 ng mL− 1 , while electrochemical transduction led to a considerable (680 times) improvement, with a LOD of 0.44 ng mL− 1 . In addition, polyaldehydedextran showed priority over polycarboxylated-dextran as the multivalent antibody crosslinker for MNPs in terms of sensitivity and LOD value, while immunosensors constructed with carboxylated magnetic microbeads (HOOC-MBs) outperformed MNPs-based immunoplatforms. This work sheds light on the importance of surface chemistry (type and density of functional groups) and the dimension (nanosize vs micrometer) of magnetic carriers to conjugate antibodies with better directional orientation and improve the analytical performance of the resulting immunosensors.
Empowering Electrochemical Biosensing through Nanostructured or Multifunctional Nucleic Acid or Peptide Biomaterials
(Advanced Materials Technologies, 2022) Campuzano Ruiz, Susana; Pedrero Muñoz, María; Barderas, Rodrigo; Pingarrón Carrazón, José Manuel
Electrochemical biosensors continue to evolve at an astonishing pace, consolidating as competitive tools for determining a wide range of targets and relentlessly strengthening their attributes in terms of sensitivity, selectivity, simplicity, response time, and antifouling ability, making them suitable for getting a foothold in real-world applications. The design and exploitation of nanostructured or multifunctional nucleic acid or peptide biomaterials is playing a determinant role in these achievements. With the aim of highlighting the potential and opportunities of these biomaterials, this perspective article critically discusses and overviews the electrochemical biosensors reported since 2019 involving nanostructured and multifunctional DNA biomaterials, multifunctional aptamers, modern peptides, and CRISPR/Cas systems. The use of these biomaterials as recognition elements, electrode modifiers (acting as linkers or creating scaffolds with antifouling properties), enzyme substrates, and labeling/carrier agents for signal amplification is discussed through rationally and strategically selected examples, concluding with a personal perspective about the challenges to be faced and future lines of action.
Determinación de herbicidas nitrofenoles y s-triazinas por voltamperometría de adsorción-redisolución
(2002) Pedrero Muñoz, María; Manuel de Villena Rueda, Francisco Javier; Pingarrón Carrazón, José Manuel
En este trabajo se han desarrollado métodos para determinar los herbicidas dinoseb, metoprotrina y terbutrina empleando polarografía diferencial de impulsos (dpp) y voltamperometria de adsorción-redisolución (adsv). A partir del comportamiento polarográfico de los tres herbicidas se han establecido las caracteristicas del proceso electrodo proponiéndose los mecanismos de reducción correspondientes y métodos para su determinación mediante dpp que se han aplicado para determinar dinoseb en manzanas y metoprotrina y terbutrina en peras. Tras verificar la posibilidad de utilizar la adsorción como etapa de preconcentración efectiva, se han desarrollado métodos adsv para determinar los tres herbicidas sobre el electrodo de gota colgante de mercurio (hmde). Se han establecido sus caracteristicas analíticas y se han aplicado a la determinación de dinoseb en manzanas y zumo de manzana y de metoprotrina y terbutrina individualmente en aguas potables y procedentes de riego. Asimismo, se ha desarrollado un método para determinar dinoseb por adsv sobre el electrodo de película de mercurio, mejorando el límite de determinación del orden de 10 veces con respecto al obtenido sobre el hmde. Este método se ha aplicado a la determinación del herbicida en muestras de zumo de manzana. Finalmente, se ha evaluado la posibilidad de aplicar la técnica adsv en medios dispersos. Determinando terbutrina en medio micelar formado con pentanosulfonato sódico y en medio emulsionado constituido con acetato de etilo se han obtenido limites de determinación del mismo orden que en disoluciones acuosas.
Electrochemical sensor for rapid determination of fibroblast growth factor receptor 4 in raw cancer cell lysates
(PLOS ONE, 2017) Khodarahmi, Reza; Torrente Rodríguez, Rebeca M.; Ruiz-Valdepeñas Montiel, Víctor; Campuzano, Susana; Pedrero Muñoz, María; Farchado, Meryem; Vargas, Eva; Manuel de Villena, F. Javier; Garranzo Asensio, María; Barderas, Rodrigo; Pingarrón Carrazón, José Manuel
The first electrochemical immunosensor for the determination of fibroblast growth factor receptor 4 (FGFR4) biomarker is reported in this work. The biosensor involves a sandwich configuration with covalent immobilization of a specific capture antibody onto activated carboxylic-modified magnetic microcarriers (HOOC-MBs) and amperometric detection at disposable carbon screen-printed electrodes (SPCEs). The biosensor exhibits a great analytical performance regarding selectivity for the target protein and a low LOD of 48.2 pg mL-1. The electrochemical platform was successfully applied for the determination of FGFR4 in different cancer cell lysates without any apparent matrix effect after a simple sample dilution and using only 2.5 μg of the raw lysate. Comparison of the results with those provided by a commercial ELISA kit shows competitive advantages by using the developed immunosensor in terms of simplicity, analysis time, and portability and cost-affordability of the required instrumentation for the accurate determination of FGFR4 in cell lysates.
Magnetic microbeads-based amperometric immunoplatform for the rapid and sensitive detection of N6-methyladenosine to assist in metastatic cancer cells discrimination
(Biosensors and Bioelectronics, 2021) Povedano Muñumel, Eloy; Gamella Carballo, María; Torrente Rodríguez, Rebeca Magnolia; Montero-Calle, Ana; Pedrero Muñoz, María; Solís-Fernández, Guillermo; Navarro Villoslada, Fernando; Barderas, Rodrigo; Campuzano Ruiz, Susana; Pingarrón, José; Pingarrón Carrazón, José Manuel
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.
Beyond Sensitive and Selective Electrochemical Biosensors: Towards Continuous, Real-Time, Antibiofouling and Calibration-Free Devices
(Sensors, 2020) Campuzano Ruiz, Susana; Pedrero Muñoz, María; Gamella Carballo, Maria; Serafín González-Carrato, Verónica; Yáñez Sedeño, Paloma; Pingarrón Carrazón, José Manuel
Nowadays, electrochemical biosensors are reliable analytical tools to determine a broad range of molecular analytes because of their simplicity, affordable cost, and compatibility with multiplexed and point-of-care strategies. There is an increasing demand to improve their sensitivity and selectivity, but also to provide electrochemical biosensors with important attributes such as near real-time and continuous monitoring in complex or denaturing media, or in vivo with minimal intervention to make them even more attractive and suitable for getting into the real world. Modification of biosensors surfaces with antibiofouling reagents, smart coupling with nanomaterials, and the advances experienced by folded-based biosensors have endowed bioelectroanalytical platforms with one or more of such attributes. With this background in mind, this review aims to give an updated and general overview of these technologies as well as to discuss the remarkable achievements arising from the development of electrochemical biosensors free of reagents, washing, or calibration steps, and/or with antifouling properties and the ability to perform continuous, real-time, and even in vivo operation in nearly autonomous way. The challenges to be faced and the next features that these devices may offer to continue impacting in fields closely related with essential aspects of people’s safety and health are also commented upon.
Antifouling (Bio)materials for Electrochemical (Bio)sensing
(International Journal of Molecular Sciences, 2019) Campuzano Ruiz, Susana; Pedrero Muñoz, María; Yáñez Sedeño, Paloma; Pingarrón Carrazón, José Manuel
(Bio)fouling processes arising from nonspecific adsorption of biological materials (mainly proteins but also cells and oligonucleotides), reaction products of neurotransmitters oxidation, and precipitation/polymerization of phenolic compounds, have detrimental effects on reliable electrochemical (bio)sensing of relevant analytes and markers either directly or after prolonged incubation in rich-proteins samples or at extreme pH values. Therefore, the design of antifouling (bio)sensing interfaces capable to minimize these undesired processes is a substantial outstanding challenge in electrochemical biosensing. For this purpose, efficient antifouling strategies involving the use of carbon materials, metallic nanoparticles, catalytic redox couples, nanoporous electrodes, electrochemical activation, and (bio)materials have been proposed so far. In this article, biomaterial-based strategies involving polymers, hydrogels, peptides, and thiolated self-assembled monolayers are reviewed and critically discussed. The reported strategies have been shown to be successful to overcome (bio)fouling in a diverse range of relevant practical applications. We highlight recent examples for the reliable sensing of particularly fouling analytes and direct/continuous operation in complex biofluids or harsh environments. Opportunities, unmet challenges, and future prospects in this field are also pointed out.
Magnetic Beads-Based Sensor with Tailored Sensitivity for Rapid and Single-Step Amperometric Determination of miRNAs
(International Journal of Molecular Sciences, 2017) Vargas Orgaz, Eva; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Povedano Muñumel, Eloy; Pedrero Muñoz, María; Montoya, Juan; Campuzano Ruiz, Susana; Pingarrón Carrazón, José Manuel
This work describes a sensitive amperometric magneto-biosensor for single-step and rapid determination of microRNAs (miRNAs). The developed strategy involves the use of direct hybridization of the target miRNA (miRNA-21) with a specific biotinylated DNA probe immobilized on streptavidin-modified magnetic beads (MBs), and labeling of the resulting heteroduplexes with a specific DNA–RNA antibody and the bacterial protein A (ProtA) conjugated with an horseradish peroxidase (HRP) homopolymer (Poly-HRP40) as an enzymatic label for signal amplification. Amperometric detection is performed upon magnetic capture of the modified MBs onto the working electrode surface of disposable screen-printed carbon electrodes (SPCEs) using the H2O2/hydroquinone (HQ) system. The magnitude of the cathodic signal obtained at −0.20 V (vs. the Ag pseudo-reference electrode) demonstrated linear dependence with the concentration of the synthetic target miRNA over the 1.0 to 100 pM range. The method provided a detection limit (LOD) of 10 attomoles (in a 25 μL sample) without any target miRNA amplification in just 30 min (once the DNA capture probe-MBs were prepared). This approach shows improved sensitivity compared with that of biosensors constructed with the same anti-DNA–RNA Ab as capture instead of a detector antibody and further labeling with a Strep-HRP conjugate instead of the Poly-HRP40 homopolymer. The developed strategy involves a single step working protocol, as well as the possibility to tailor the sensitivity by enlarging the length of the DNA/miRNA heteroduplexes using additional probes and/or performing the labelling with ProtA conjugated with homopolymers prepared with different numbers of HRP molecules. The practical usefulness was demonstrated by determination of the endogenous levels of the mature target miRNA in 250 ng raw total RNA (RNAt) extracted from human mammary epithelial normal (MCF-10A) and cancer (MCF-7) cells and tumor tissues.
Non-Invasive Breast Cancer Diagnosis through Electrochemical Biosensing at Different Molecular Levels
(Sensors, 2017) Campuzano, Susana; Pedrero Muñoz, María; Pingarrón Carrazón, José Manuel
The rapid and accurate determination of specific circulating biomarkers at different molecular levels with non- or minimally invasive methods constitutes a major challenge to improve the breast cancer outcomes and life quality of patients. In this field, electrochemical biosensors have demonstrated to be promising alternatives against more complex conventional strategies to perform fast, accurate and on-site determination of circulating biomarkers at low concentrations in minimally treated body fluids. In this article, after discussing briefly the relevance and current challenges associated with the determination of breast cancer circulating biomarkers, an updated overview of the electrochemical affinity biosensing strategies emerged in the last 5 years for this purpose is provided highlighting the great potentiality of these methodologies. After critically discussing the most interesting features of the electrochemical strategies reported so far for the single or multiplexed determination of such biomarkers with demonstrated applicability in liquid biopsy analysis, existing challenges still to be addressed and future directions in this field will be pointed out.