Person: Benito Peña, María Elena
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First Name
María Elena
Last Name
Benito Peña
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Químicas
Department
Química Analítica
Area
Química Analítica
Identifiers
18 results
Search Results
Now showing 1 - 10 of 18
Item Molecular super-gluing: a straightforward tool for antibody labelling and its application to mycotoxin biosensing(Analytical and Bioanalytical Chemistry, 2022) Prádanas González, Fernando; Glahn Martínez, Ana Bettina; Benito Peña, María Elena; Arola, Henri O.; Nevanen, Tarja K.; Moreno Bondi, María CruzMycotoxins are low molecular weight toxic compounds, which can cause severe health problems in animals and humans. Immunoassays allow rapid, simple and cost-efective screening of mycotoxins. Sandwich assays with a direct readout provide great improvement in terms of selectivity and sensitivity, compared to the widely used competitive assay formats, for the analysis of low molecular weight molecules. In this work, we report a non-competitive fuorescence anti-immune complex (IC) immunoassay, based on the specifc recognition of HT-2 toxin with a pair of recombinant antibody fragments, namely antigen-binding fragment (Fab) (anti-HT-2 (10) Fab) and single-chain variable fragment (scFv) (anti-IC HT-2 (10) scFv). The SpyTag and SpyCatcher glue proteins were applied for the frst time as a bioconjugation tool for the analysis of mycotoxins. To this aim, a SpyTag-mScarlet-I (fuorescent protein) and scFv-SpyCatcher fusion proteins were constructed, produced and fused in situ during the assay by spontaneous Tag-Catcher binding. The assay showed an excellent sensitivity with an EC50 of 4.8±0.4 ng mL−1 and a dynamic range from 1.7±0.3 to 13±2 ng mL−1, an inter-day reproducibility of 8.5% and a high selectivity towards HT-2 toxin without cross-reactivity with other Fusarium toxins. The bioassay was applied to the analysis of the toxin in an oat reference material and in oat samples, with a LOD of 0.6 µg kg−1, and the results were validated by analysing a certifcate reference material and by HPLC–MS/MS.- Project number: 253
Item Material docente interactivo en inglés para la enseñanza práctica y el autoaprendizaje de (bio)sensores químicos ópticos en Grado y Máster(2018) Descalzo López, Ana Belén; Orellana Moraleda, Guillermo; Moreno Bondi, María Cruz; Benito Peña, María Elena; Urraca Ruiz, JavierMaterial docente (guiones de prácticas, vídeos, cuestionarios multi-respuesta) en inglés para asignaturas de Grado y Máster relacionadas con el tema de sensores (bio)químicos ópticos y la preparación de nanomateriales aplicados en sensores ópticos Item Comparative Study of the Performance of Two Different Luciferases for the Analysis of Fumonisin B1 in Wheat Samples(Analysis & Sensing, 2022) Luque Uría, Álvaro; Peltomaa, Riikka; Navarro Duro, Marina; Fikacek, Sabrina; Head, Trajen; Deo, Sapna K.; Daunert, Sylvia; Benito Peña, María Elena; Moreno Bondi, María CruzThe development of two different immunoassays for the determination of fumonisin B1 in wheat samples is reported. A previously described mimopeptide for fumonisin B1 (FB1) was used to produce fusion proteins in combination with two different luciferases: Gaussia luciferase (GLuc) and NanoLuc luciferase (NLuc). The production, expression and the development of two immunoassays based on these fusion proteins (A2- GLuc and A2-NLuc) is detailed. The assay showing the best performance, A2-NLuc, with a limit of detection of 0.61 ngmL 1 and a dynamic range from 1.9 to 95 ngmL 1 , was employed for the analysis of spiked wheat samples, a reference matrix material, as well as naturally contaminated wheat samples. The recoveries obtained in the spiked samples were acceptable, between 81.5 and 109%, with relative standard deviations lower than 14%. The analysis of naturally contaminated wheat was validated by a liquid chromatography coupled to tandem mass detection method.Item Homogeneous quenching immunoassay for fumonisin B1 based on gold nanoparticles and an epitope-mimicking yellow fluorescent protein(ACS Nano, 2018) Peltomaa, Riikka Johanna; Amaro Torres, Francisco; Carrasco, Sergio; Orellana Moraleda, Guillermo; Benito Peña, María Elena; Moreno Bondi, María CruzHomogeneous immunoassays represent an attractive alternative to traditional heterogeneous assays due to their simplicity, sensitivity, and speed. On the basis of a previously identified epitope-mimicking peptide, or mimotope, we developed a homogeneous fluorescence quenching immunoassay based on gold nanoparticles (AuNPs) and a recombinant epitope-mimicking fusion protein for the detection of mycotoxin fumonisin B1 (FB1). The fumonisin mimotope was cloned as a fusion protein with a yellow fluorescent protein that could be used directly as the tracer for FB1 detection without the need of labeling or a secondary antibody. Furthermore, owing to the fluorescence quenching ability of AuNPs, a homogeneous immunoassay could be performed in a single step without washing steps to separate the unbound tracer. The homogeneous quenching assay showed negligible matrix effects in 5% wheat extract and high sensitivity for FB1 detection, with a dynamic range from 7.3 to 22.6 ng/mL, a detection limit of 1.1 ng/mL, and IC50 value of 12.9 ng/mL, which was significantly lower than the IC50 value of the previously reported assay using the synthetic counterpart of the same mimotope in a microarray format. The homogeneous assay was demonstrated to be specific for fumonisins B1 and B2, as no significant cross-reactivity with other mycotoxins was observed, and acceptable recoveries (86% for FB1 2000 μg/kg and 103% for FB1 4000 μg/kg), with relative standard deviation less than 6.5%, were reported from spiked wheat samples, proving that the method could provide a valuable tool for simple analysis of mycotoxin-contaminated food samples.- Project number: 179
Item "Chem-game", el juego como estrategia para la dinamización del aprendizaje y la evaluación de conocimientos en Química General(2020) Moreno Bondi, María Cruz; Gutiérrez Alonso, Angel; Lainez Ferrando, Alfredo; Benito Peña, María Elena; Glahn Martínez, Ana Bettina; García Pinar, Gioele; Castellano Perdomo, Raúl; Navarro Duro, Marina; Peng, Wenrong; Morales Benítez, Noelia; Rubio Ramón, Pablo Ignacio; Olmos Alonso, InésEste proyecto pretende aplicar la "gamificación" en la enseñanza de la asignatura de Química General de primer curso del grado en Química para fomentar la formación, creatividad, compromiso y la capacidad de trabajo en equipo de los estudiantes. Item Biosensing Based on Nanoparticles for Food Allergens Detection(Sensors, 2018) Gómez Arribas, Lidia Nazaret; Benito Peña, María Elena; Hurtado Sánchez, María del Carmen; Moreno Bondi, María CruzFood allergy is one of the major health threats for sensitized individuals all over the world and, over the years, the food industry has made significant efforts and investments to offer safe foods for allergic consumers. The analysis of the concentration of food allergen residues in processing equipment, in raw materials or in the final product, provides analytical information that can be used for risk assessment as well as to ensure that food-allergic consumers get accurate and useful information to make their food choices and purchasing decisions. The development of biosensors based on nanomaterials for applications in food analysis is a challenging area of growing interest in the last years. Research in this field requires the combined efforts of experts in very different areas including food chemistry, biotechnology or materials science. However, the outcome of such collaboration can be of significant impact on the food industry as well as for consumer’s safety. These nanobiosensing devices allow the rapid, selective, sensitive, cost-effective and, in some cases, in-field, online and real-time detection of a wide range of compounds, even in complex matrices. Moreover, they can also enable the design of novel allergen detection strategies. Herein we review the main advances in the use of nanoparticles for the development of biosensors and bioassays for allergen detection, in food samples, over the past few years. Research in this area is still in its infancy in comparison, for instance, to the application of nanobiosensors for clinical analysis. However, it will be of interest for the development of new technologies that reduce the gap between laboratory research and industrial applications.Item Extracting mycotoxins from edible vegetable oils by using green, ecofriendly deep eutectic solvents(Food Chemistry, 2023) Pradanas-González, Fernando; Aragoneses-Cazorla, Rubén; Ángel Merino-Sierra, Miguel; Andrade-Bartolomé, Elena; Navarro-Villoslada, Fernando; Benito-Peña, Elena; Moreno-Bondi, María Cruz; Moreno Bondi, María Cruz; Benito Peña, María Elena; Navarro Villoslada, Fernando; Prádanas González, FernandoIn this work, we developed an environmentally friendly liquid–liquid microextraction method using a natural deep eutectic solvent in combination with liquid chromatography for the simultaneous determination of four mycotoxins (deoxynivalenol, alternariol, ochratoxin A and zearalenone) in edible vegetable oils. A chemometric approach assessed the effect of the operational parameters on the mycotoxin extraction efficiency. The extracts were analyzed by HPLC coupled with a diode array and fluorescence detector. The optimum NADES composition resulted in the highest extraction recoveries, and it was applied to coextract the target mycotoxins in several types of edible vegetable oils without using hazardous solvents or requiring further clean-up. The limits of detection ranged from 0.07 to 300μg/kg, and recoveries were close to 100%, except for zearalenone(viz. 35%), with relative standard deviations below 9% in all cases. The proposed method was validated following the European Commission 2002/657/EC and 2006/401/EC.Item Magnetic Janus micromotors for fluorescence biosensing of tacrolimus in oral fluids(Biosensors and Bioelectronics, 2024) Glahn Martínez, Ana Bettina; Jurado-Sánchez, Beatriz; Benito Peña, María Elena; Escarpa, Alberto; Moreno Bondi, María CruzTacrolimus (FK506) is a macrolide lactone immunosuppressive drug that is commonly used in transplanted patients to avoid organ rejection. FK506 exhibits high inter- and intra-patient pharmacokinetic variability, making monitoring necessary for organ graft survival. This work describes the development of a novel bioassay for monitoring FK506. The bioassay is based on using polycaprolactone-based (PCL) magnetic Janus micromotors and a recombinant chimera receptor that incorporates the immunophilin tacrolimus binding protein 1A (FKBP1A) tagged with Emerald Green Fluorescent Protein (EmGFP). The approach relies on a fluorescence competitive bioassay between the drug and the micromotors decorated with a carboxylated FK506 toward the specific site of the fluorescent immunophilin. The proposed homogeneous assay could be performed in a single step without washing steps to separate the unbound receptor. The proposed approach fits the therapeutic requirements, showing a limit of detection of 0.8 ng/mL and a wide dynamic range of up to 90 ng/mL. Assay selectivity was evaluated by measuring the competitive inhibition curves with other immunosuppressive drugs usually co-administered with FK506. The magnetic propulsion mechanism allows for efficient operation in raw samples without damaging the biological binding receptor (FKBP1A-EmGFP). The enhanced target recognition and micromixing strategies hold considerable potential for FK506 monitoring in practical clinical use.Item Highly fluorescent magnetic nanobeads with a remarkable stokes shift as labels for enhanced detection in immunoassays(Small, 2018) Salis, Francesca; Descalzo López, Ana Belén; Benito Peña, María Elena; Moreno Bondi, María Cruz; Orellana Moraleda, GuillermoFluorescence immunoassays are popular for achieving high sensitivity, but they display limitations in biological samples due to strong absorption of light, background fluorescence from matrix components, or light scattering by the biomacromolecules. A powerful strategy to overcome these problems is introduced here by using fluorescent magnetic nanobeads doped with two boron-dipyrromethane dyes displaying intense emission in the visible and near-infrared regions, respectively. Careful matching of the emission and absorption features of the dopants leads to a virtual Stokes shift larger than 150 nm achieved by an intraparticle Förster resonance energy transfer (FRET) process between the donor and the acceptor dyes. Additionally, the magnetic properties of the fluorescent beads allow preconcentration of the sample. To illustrate the usefulness of this approach to increase the sensitivity of fluorescence immunoassays, the novel nanoparticles are employed as labels for quantification of the widely used Tacrolimus (FK506) immunosuppressive drug. The FRET-based competitive inhibition immunoassay yields a limit of detection (LOD) of 0.08 ng/mL, with a dynamic range (DR) of 0.15–2.0 ng/mL, compared to a LOD of 2.7 ng/mL and a DR between 4.1 and 130 ng/mL for the immunoassay carried out with direct excitation of the acceptor dye.Item Homogeneous immunoassay for cyclopiazonic acid based upon mimotopes and upconversion-resonance energy transfer(Biosensors and Bioelectronics, 2023) Prádanas González, Fernando; Peltomaa, Riikka Johanna; Satu Lahtinen; Luque Uria, Álvaro; Vicente Más; Rodrigo Barderas; Chris M. Maragos; Canales Mayordomo, María Ángeles; Tero Soukka; Benito Peña, María Elena; Moreno Bondi, María CruzStrains of Penicillium spp. are used for fungi-ripened cheeses and Aspergillus spp. routinely contaminate maize and other crops. Some of these strains can produce toxic secondary metabolites (mycotoxins), including the neurotoxin α-cyclopiazonic acid (CPA). In this work, we developed a homogeneous upconversion-resonance energy transfer (UC-RET) immunoassay for the detection of CPA using a novel epitope mimicking peptide, or mimotope, selected by phage display. CPA-specific antibody was used to isolate mimotopes from a cyclic 7-mer peptide library in consecutive selection rounds. Enrichment of antibody binding phages was achieved, and the analysis of individual phage clones revealed four different mimotope peptide sequences. The mimotope sequence, ACNWWDLTLC, performed best in phage-based immunoassays, surface plasmon resonance binding analyses, and UC-RET-based immunoassays. To develop a homogeneous assay, upconversion nanoparticles (UCNP, type NaYF4:Yb3+, Er3+) were used as energy donors and coated with streptavidin to anchor the synthetic biotinylated mimotope. Alexa Fluor 555, used as an energy acceptor, was conjugated to the anti-CPA antibody fragment. The homogeneous single-step immunoassay could detect CPA in just 5 min and enabled a limit of detection (LOD) of 30 pg/mL (1.5 μg/kg) and an IC50 value of 0.36 ng/mL. No significant cross-reactivity was observed with other co-produced mycotoxins. Finally, we applied the novel method for the detection of CPA in spiked maize samples using high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD) as a reference method.