Optical Biosensors for Label-Free Detection of Small Molecules

Label-free optical biosensors are an intriguing option for the analyses of many analytes, as they offer several advantages such as high sensitivity, direct and real-time measurement in addition to multiplexing capabilities. However, development of label-free optical biosensors for small molecules can be challenging as most of them are not naturally chromogenic or fluorescent, and in some cases, the sensor response is related to the size of the analyte. To overcome some of the limitations associated with the analysis of biologically, pharmacologically, or environmentally relevant compounds of low molecular weight, recent advances in the field have improved the detection of these analytes using outstanding methodology, instrumentation, recognition elements, or immobilization strategies. In this review, we aim to introduce some of the latest developments in the field of label-free optical biosensors with the focus on applications with novel innovations to overcome the challenges related to small molecule detection. Optical label-free methods with different transduction schemes, including evanescent wave and optical fiber sensors, surface plasmon resonance, surface-enhanced Raman spectroscopy, and interferometry, using various biorecognition elements, such as antibodies, aptamers, enzymes, and bioinspired molecularly imprinted polymers, are reviewed

Biosensing Based on Nanoparticles for Food Allergens Detection

Food 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

Comparative Study of the Performance of Two Different Luciferases for the Analysis of Fumonisin B1 in Wheat Samples

The 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

Aluminum Nanoholes for Optical Biosensing

[EN] Sub-wavelength diameter holes in thin metal layers can exhibit remarkable optical features that make them highly suitable for (bio)sensing applications. Either as efficient light scattering centers for surface plasmon excitation or metal-clad optical waveguides, they are able to form strongly localized optical fields that can effectively interact with biomolecules and/or nanoparticles on the nanoscale. As the metal of choice, aluminum exhibits good optical and electrical properties, is easy to manufacture and process and, unlike gold and silver, its low cost makes it very promising for commercial applications. However, aluminum has been scarcely used for biosensing purposes due to corrosion and pitting issues. In this short review, we show our recent achievements on aluminum nanohole platforms for (bio)sensing. These include a method to circumvent aluminum degradation-which has been successfully applied to the demonstration of aluminum nanohole array (NHA) immunosensors based on both, glass and polycarbonate compact discs supports-the use of aluminum nanoholes operating as optical waveguides for synthesizing submicron-sized molecularly imprinted polymers by local photopolymerization, and a technique for fabricating transferable aluminum NHAs onto flexible pressure-sensitive adhesive tapes, which could facilitate the development of a wearable technology based on aluminum NHAs.The authors gratefully acknowledge financial support from MINECO projects, Spain (TEC2012-31145, CTQ2012-37573-C02 and CTQ 2013-45875-R).Angulo Barrios, C.; Canalejas Tejero, V.; Herranz, S.; Urraca, J.; Moreno-Bondi, MC.; Avella-Oliver, M.; Maquieira Catala, Á.... (2015). Aluminum Nanoholes for Optical Biosensing. Biosensors. 5(3):417-431. https://doi.org/10.3390/bios50304174174315

"Chem-game", el juego como estrategia para la dinamización del aprendizaje y la evaluación de conocimientos en Química General

Este 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

Diseño y preparación de un laboratorio virtual de Química Analítica: Técnicas instrumentales de análisis

La reciente pandemia del COVID-19 ha supuesto un cambio excepcional y drástico de la concepción tradicional del aprendizaje, tanto para los estudiantes como para los docentes. Ante esta situación se requiere no sólo acciones que faciliten la adaptación de los estudiantes y profesores a las plataformas educativas en línea, sino también a que éstas se conviertan en auténticas herramientas para potenciar y mejorar de forma significativa el aprendizaje del alumno. El presente proyecto de Innovación Educativa y Mejora de la Calidad Docente pretende mejorar la calidad del aprendizaje de varias asignaturas de los Grados de Química e Ingeniería Química que llevan asociado un Laboratorio de Técnicas Instrumentales. Tradicionalmente esa docencia práctica ha sido presencial, pero la situación excepcional surgida con la pandemia en el curso 2019/2020, ha demostrado que disponer de unas prácticas virtualizadas es de gran ayuda para facilitar el aprendizaje de los alumnos y facilitar una transferencia del conocimiento constructivo y colaborativo. Desde hace años, el personal docente e investigador (PDI), personal de administración y servicios (PAS) y los estudiantes de la UCM, disponemos de la plataforma de enseñanza online Moodle (Campus Virtual, CV). Sin embargo, la pandemia que vivimos ha evidenciado la brecha digital en lo referente a cómo usar Moodle y, por ende, la utilización de las Tecnologías de Información y Comunicación (TICs) como facilitadoras didácticas. A pesar de nuestras limitaciones, tanto profesores como estudiantes, hemos desarrollado una enorme capacidad resiliente, lo que permitió, durante el pasado mes de mayo, la puesta en marcha de los primeros laboratorios en línea en el Departamento de Química Analítica de la UCM. Fruto de esta experiencia, así como de la situación actual de incertidumbre para el curso próximo, algunos profesores, estudiantes y PAS del departamento hemos decidido adelantarnos a un escenario futuro en el que se contemple nuevamente la impartición de Docencia Experimental en línea y solicitar el presente proyecto de Innovación Educativa y Mejora de la Calidad Docente

Tag-specific affinity purification of recombinant proteins by using molecularly imprinted polymers

Epitope tagging is widely used to fuse a known epitope to proteins for which no affinity receptor is available by using recombinant DNA technology. One example is FLAG epitope (DYKDDDDK), which provides better purity and recoveries than the favorite polyhistidine tag. However, purification requires using anti-FLAG antibody resins, the high cost and non-reusability of which restrict widespread use. One cost-effective solution is provided by the use of bioinspired anti-FLAG molecularly imprinted polymers (MIPs). This work describes the development of MIPs, based on the epitope approach, synthesized from the tetrapeptide DYKD as template that affords purification of FLAG-derived recombinant proteins. Polymer was optimized by using a combinatorial approach to select the functional monomer(s) and cross-linker(s), resulting in the best specific affinity toward FLAG and the peptide DYKD. The imprinted resin obtained was used to purify mCherry proteins tagged with either FLAG or DYKD epitopes from crude cell lysates. Both mCherry variants were highly efficiently purified (R ≥ 95%, RSD ≤ 15%, n = 3) and impurities were removed. Unlike existing antibody-based resins, the proposed tag-imprinting strategy provides a general method for meeting the growing demand for efficient, inexpensive, and versatile materials for tagged proteins purification.Spanish Ministry of Economy and CompetitivenessDepto. de Química AnalíticaFac. de Ciencias QuímicasFALSEpu

Biosensing Based on Nanoparticles for Food Allergens Detection

Food 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