Person:
Moreno Guzmán, María

First Name

María

Last Name

Moreno Guzmán

URI

https://hdl.handle.net/20.500.14352/77485

Affiliation

Universidad Complutense de Madrid

Faculty / Institute

Farmacia

Department

Química en Ciencias Farmacéuticas

Area

Química Analítica

Identifiers

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Search Results

Now showing 1 - 10 of 11

Paving the Way for Reliable Alzheimer’s Disease Blood Diagnosis by Quadruple Electrochemical Immunosensing
(ChemElectroChem, 2022) Valverde de la Fuente, Alejandro; Gordón Pidal, José María; Montero Calle, Ana; Arévalo Pérez, Beatriz; Serafín González-Carrato, Verónica; Calero, Miguel; Moreno Guzmán, María; López, Miguel Ángel; Escarpa, Alberto; Yáñez Sedeño, Paloma; Barderas, Rodrigo; Campuzano Ruiz, Susana; Pingarrón Carrazón, José Manuel
Alzheimer’s disease (AD), the most common neurodegenerative disorder, demands new cost-effective and easy-to-use strategies for its reliable detection, mainly in the preclinical stages. Here, we report the first immunoplatform for the electrochemical multidetermination of four candidate protein biomarkers in blood, neurofilament light chain (NfL), Tau, phosphorylated Tau (p-Tau) and TAR DNA-Binding Protein 43 (TDP-43). It involves implementation of sandwich-type immunoassays and enzymatic labelling with horseradish peroxidase (HRP) on the surface of magnetic microbeads (MBs). Amperometric detection is performed after depositing the magnetic immunoconjugates on disposable quadruple transduction platforms by monitoring the enzymatic reduction of H2O2 mediated by hydroquinone (HQ). The immunoplatform achieved LOD values smaller than the content of target biomarkers in plasma of healthy subjects, with RSD values.
An array-based electrochemical magneto-immunosensor for early neonatal sepsis diagnostic: Fast and accurate determination of C-reactive protein in whole blood and plasma samples
(Microchemical Journal, 2020) Molinero-Fernández, Águeda; Moreno Guzmán, María; López, Miguel Ángel; Escarpa, Alberto
An array-based electrochemical magneto-immunosensor (a-EMI) constituted by eight screen-printed carbon electrodes is proposed for the determination of C-reactive protein (CRP). Remarkably, the analytical usefulness of the approach was demonstrated not only by analyzing whole blood samples, but plasma ones from newborns with suspicion of sepsis, which sample volume is hardly available. The a-EIM enabled the accurate determination of CRP in these unique samples (Er < 3%) with excellent intra (CV<4%, n=8) and inter-assays (CV<9%, n=8) precision, using only 5 µL of clinical sample, covering the sepsis diagnostic range and employing only 15 min for total assay. The electrode array enabled to work with up to 8 tailored dilutions simultaneously accordingly to the CRP clinical diagnostic range. It constitutes by itself a valuable advantage for speed and reliability improvements in the sample analysis and further sepsis diagnosis. These results non-only highly met the clinical needs for sepsis newborns diagnosis, but they opened new perspectives in low volumes-based diagnostics.
On the Move-Sensitive Fluorescent Aptassay on Board Catalytic Micromotors for the Determination of Interleukin-6 in Ultra-Low Serum Volumes for Neonatal Sepsis Diagnostics
(ACS Sensors, 2022) Gordón, José; Arruza Gómez, Luis; Ibáñez, María Dolores; Moreno Guzmán, María; López, Miguel Ángel; Escarpa, Alberto
A graphene oxide/nickel/platinum nanoparticle micromotor (MM)-based fluorescent aptassay is proposed to determine interleukin-6 (IL-6) in serum samples from low-birth-weight infants (gestational age of less than 32 weeks and birthweight below 1000 g) with sepsis suspicion. In this kind of patients, IL-6 has demonstrated good sensitivity and specificity for the diagnosis of sepsis, both for early and late onset sepsis. The approach was based on the adsorption of the aptamer for IL6 tagged with 6-FAM as a fluorescent label (AptIL‑6, λem = 520 nm) on the graphene oxide external layer (MMGO−AptIL‑6) inducing fluorescence quenching (OFF state) and a subsequent on-the-move affinity recognition of IL-6 from AptIL‑6 (IL-6−AptIL‑6 complex) recovering the fluorescence (ON state). An aptamer against IL-6 was selected and developed by the systematic evolution of ligands by exponential enrichment technology. This approach displayed a suitable linear range of 0.07−1000 pg mL−1 (r = 0.995) covering the cut-off and clinical practice levels, allowing direct determination without any dilution and simplifying the analysis as well as exhibiting an excellent sensitivity (LOD =0.02 pg mL−1) in ultralow volumes of diagnostic clinical samples (2 μL). A high agreement between IL-6 levels obtained from our MM-based approach and the method used by the Hospital was obtained (relative error < 3%). The MM-based aptassay is competitive in comparison with that of the Hospital, in terms of a significant reduction of the sample volume (15 times less) and enhanced sensitivity, employing similar analysis times. These results position MM technology with enough potential to achieve high sensitivities in low sample volumes, opening new avenues in diagnosis based on low sample volumes.
Toward Early Diagnosis of Late-Onset Sepsis in Preterm Neonates: Dual Magnetoimmunosensor for Simultaneous Procalcitonin and C-Reactive Protein Determination in Diagnosed Clinical Samples
(ACS Sensors, 2019) Molinero-Fernández, Águeda ; Moreno Guzmán, María; Arruza Gómez, Luis; López, Miguel Ángel; Escarpa, Alberto
Early diagnosis of sepsis, combining blood cultures and inflammation biomarkers, continues to be a challenge, especially in very low birth weight (VLBW) infants because of limited availability of blood samples. Traditional diagnostic procedures are cumbersome, not fast enough, and require relatively large volumes of sample. Empiric use of antibiotics, before diagnostic confirmation, is required to decrease mortality, leading to potential antibiotic resistance and side effects in VLBW infants. To solve such a serious problem, a dual magnetoimmunosensor is proposed for simultaneous assessment of two of the most important sepsis biomarkers: procalcitonin (PCT for early phase) and C-reactive protein (CRP for late phase). This "sample-to-result" approach exhibited excellent sensitivity, selectivity, precision, and stability using low sample volumes (<30 μL) and under 20 min of total assay. The analytical usefulness of the approach was demonstrated by analyzing clinically relevant samples of preterm neonates with suspicion of sepsis.
Polymer-Based Micromotor Fluorescence Immunoassay for "On-the-Move" Sensitive Procalcitonin Determination in Very Low Birth Weight Infants’ Plasma
(ACS Sensor, 2020) Molinero-Fernández, Águeda; Moreno Guzmán, María; Arruza Gómez, Luis; López, Miguel Ángel; Escarpa, Alberto
A new fluorescence micromotor-based immunoassay (FMIm) has been developed for procalcitonin (PCT) determination as an early sepsis diagnostic analytical tool. The micromotors combine the high binding capacity of the specific antibodies onto their polymeric polypyrrole outer layer (PPy layer), with their magnetic guidance (Ni layer) and self-propulsion by catalytic generation of oxygen bubbles (PtNP inner layer) to actively recognize the PCT antigen. This FMIm allowed a sensitive (LOD = 0.07 ng mL-1) and direct PCT determination in clinical samples from very low-birth-weight infants (VLBWI) with sepsis suspicion, using small volumes of sample (25 μL) in a clinically relevant range of concentrations (0.5-150 ng mL-1). The good agreement between PCT levels obtained by our micromotor-based method and routine immunofluorescence hospital determination demonstrates the feasibility for the analysis in VLBWI samples and its potential as a point-of-care diagnostic tool for sepsis.
Magnetic Bead-Based Electrochemical Immunoassays On-Drop and On-Chip for Procalcitonin Determination: Disposable Tools for Clinical Sepsis Diagnosis
(Biosensors, 2020) Molinero-Fernández, Águeda; Moreno Guzmán, María; López, Miguel Ángel; Escarpa, Alberto
Procalcitonin (PCT) is a known protein biomarker clinically used for the early stages of sepsis diagnosis and therapy guidance. For its reliable determination, sandwich format magnetic bead-based immunoassays with two different electrochemical detection approaches are described: (i) disposable screen-printed carbon electrodes (SPE-C, on-drop detection); (ii) electro-kinetically driven microfluidic chips with integrated Au electrodes (EMC-Au, on-chip detection). Both approaches exhibited enough sensitivity (limit of detection (LOD) of 0.1 and 0.04 ng mL−1 for SPE-C and EMC-Au, respectively; cutoff 0.5 ng mL−1), an adequate working range for the clinically relevant concentrations (0.5–1000 and 0.1–20 ng mL−1 for SPE-C and EMC-Au, respectively), and good precision (RSD < 9%), using low sample volumes (25 µL) with total assay times less than 20 min. The suitability of both approaches was successfully demonstrated by the analysis of human serum and plasma samples, for which good recoveries were obtained (89–120%). Furthermore, the EMC-Au approach enabled the easy automation of the process, constituting a reliable alternative diagnostic tool for on-site/bed-site clinical analysis.
Aligned copper nanowires as a cut-and-paste exclusive electrochemical transducer for free-enzyme highly selective quantification of intracellular hydrogen peroxide in cisplatin-treated cells
(Biosensors and Bioelectronics, 2017) García-Carmona, Laura; Moreno Guzmán, María; Martín, Aida; Benito Martínez, Selma; Fernández-Martínez, Ana B.; González, María Cristina; Lucio-Cazaña, Javier; Escarpa, Alberto
The role and reliable quantification of intracellular hydrogen peroxide during cancer therapy constitutes an unexplored and fascinating application. In this work, we report the fabrication of vertically aligned copper nanowires (v-CuNWs) using electrosynthesis on templates, and their application as a cut-and-paste exclusive and flexible electrochemical transducer. This easily adaptable electrodic platform is demonstrated for a fast, simple and free-enzyme selective quantification of intracellular hydrogen peroxide in Cisplatin-treated human renal HK-2 cells. The v-CuNWs sensor was compared with an HRP-enzyme-based biosensor showing excellent correlation and indicates the good selectivity and analytical performance of the v-CuNWs. This sensing approach opens novel avenues for monitoring cell death processes and shows the potential of H2O2 as a cellular damage biomarker, with a clear potency for further developments for in vitro diagnosis and its implication in cancer therapy.
Class enzyme-based motors for “on the fly” enantiomer analysis of amino acids
(Biosensors and Bioelectronics, 2017) García-Carmona, Laura; Moreno Guzmán, María; González, María Cristina; Escarpa, Alberto
Here, two class-enzyme motors are properly designed allowing the rapid dispersion of the class-enzyme D-amino acid oxidase (DAO) and L-amino acid oxidase (LAO) for selective “on the fly” biodetection of D and L-amino acids (AAs), respectively. The efficient movement together with the continuous release of fresh class-enzyme leads to a greatly accelerated enzymatic reaction processes without the need of external stirring or chemical and physical attachment of the enzyme. Ultra-fast detection (<2 min) and accurate quantifications of L-phenylalanine (L-Phe) in plasma and whole-blood newborns samples diagnosed with Phenylketonuria and total D-AAs in Vibrio cholera cultures are pioneer illustrated as relevant examples of each enantiomer determination. These results opens clearly novel avenues in biosensing for fast screening diagnostics, decentralized monitoring and design of future points of care.
OFF-ON on-the-fly aptassay for rapid and accurate determination of procalcitonin in very low birth weight infants with sepsis suspicion
(Sensors and Actuators B: Chemical, 2023) Gordón Pidal, José M.; Arruza Gómez, Luis; Moreno Guzmán, María; López, Miguel Ángel; Escarpa, Alberto
A fast and reliable OFF-ON micromotors (MM)-based aptassay for rapid, sensitive, and accurate determination of procalcitonin (PCT) as one of the leading neonatal sepsis biomarkers in very low birth weight infants’ serum samples is proposed. MM were composed of three specific functional layers: graphene oxide (GO) as sensing layer, Ni for magnetic guidance and platinum nanoparticles (PtNPs) as catalytic layer for self-propulsion (MMGO/Ni/PtNPs). The OFF-ON strategy relies on the binding of the aptamer tagged with alexa-405 as fluorescent label (AptPCT, λem=447 nm) onto the GO outer layer of the MM (GOMM) to induce its fluorescence quenching (GOMM-AptPCT, OFF state); which is next recovered due to selectively binding of the aptamer to the PCT and their separation from the GOMM surface (PCT-AptPCT, ON state). The fast OFF-ON on-the-fly aptassay allowed in just 5 min a highly selective and sensitive (LOD = 0.01 ng mL−1) PCT determination in clinical samples from very low birth weight infants with sepsis suspicion, using only 25 µL of sample in a clinically relevant range of concentrations (0.03–1280 ng mL−1). The high concordance between the PCT levels obtained by our MMGO/Ni/PtNPs-based aptassay and those obtained by the Hospital, demonstrated the analytical potency of our approach for the analysis of neonatal samples as well as its potential to be used as a pre-diagnostic tool for sepsis.
Bi-enzymatic biosensor for on-site, fast and reliable electrochemical detection of relevant D-amino acids in bacterial samples
(Sensors and Actuators B, 2017) Moreno Guzmán, María; García-Carmona, Laura; Molinero-Fernández, Águeda; Cava, Felipe; López Gil, Miguel Ángel; Escarpa, Alberto
In this work, a bi-enzymatic biosensor allowed the total content of D-amino acids (DAAs) determination in highly relevant matrices involving bacteria. The strategy is based on the unique coimmobilization of D-amino acid oxidase (DAAO) and horseradish peroxidase (HRP) enzymes onto a multi-walled carbon nanotubes (MWCNTs) and gold nanoparticles (AuNPs) modified screen-printed electrode (SPCE). The greater amount of AuNPs deposited and hence the greater loading of both enzymes was observed when they were deposited after the activation of the carboxylated MWCNTs with EDC/Sulfo-NHS chemistry. These platforms provided a fast (300 s) and selective quantification of DAAs with excellent precision (RSD < 5%) and accuracy (Recoveries 100–104%) in bacterial samples. Collectively, the electrochemical bi-enzymatic biosensor become an universal, fast, sensitive and easy-to-use approach to determine total content of DAAs in complex matrices.