Publications by authors named "Elisa Michelini"

Trigger valves are fundamental features in capillary-driven microfluidic systems that stop fluid at an abrupt geometric expansion and release fluid when there is flow in an orthogonal channel connected to the valve. The concept was originally demonstrated in closed-channel capillary circuits. We show here that trigger valves can be successfully implemented in open channels.

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  • * To address these challenges, researchers developed a low-cost 3D-printed microtissue device that can be used for bioassays to monitor important molecular pathways like hypoxia signaling and the p53 pathway.
  • * The study demonstrated that genetically engineered human liver and kidney cells on this platform could effectively monitor cellular responses to different treatments, indicating its potential for sustainable and efficient drug screening and precision medicine applications.
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In this work, a chemiluminescent sensing paper has been developed using a peroxidase biomimetic metal-organic framework as a versatile host platform. For the first time, we have explored the use of in situ growth of Prussian Blue nanoparticles (PB-NPs) onto the MIL-101(Fe) structure for the assembly of a ready-to-use sensing paper. In situ growth of PB-NPs has been performed on the surface of the MIL-101(n) family.

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  • Trigger valves are essential components in microfluidic systems that control fluid flow by stopping it at expansions and allowing release through connected channels.
  • This study successfully implements trigger valves in open channels and demonstrates how multiple valves can create layered capillary flow alongside main channels.
  • A model for flow dynamics at these valves was developed and validated, with implications for applications in biosensing, sample preparation, hydrogel patterning, and organ-on-a-chip technologies.
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  • * This study introduces a new immunoassay that uses a split-nanoluciferase system and nanobodies to detect PSA, enhancing sensitivity and specificity.
  • * The new method effectively detects PSA in a range from 1.0 to 20.0 ng/mL with a limit of detection of 0.4 ng/mL, showing good agreement with traditional ELISA results, making it a quick and user-friendly testing option.
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Several organ-on-chip and cell-on-chip devices have been reported, however, their main drawback is that they are not interoperable (i.e., they have been fabricated with customized equipment, thus cannot be applied in other facilities, unless having the same setup), and require cell-culture facilities and benchtop instrumentation.

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Introduction: Bioluminescence is a well-established optical detection technique widely used in several bioanalytical applications, including high-throughput and high-content screenings. Thanks to advances in synthetic biology techniques and deep learning, a wide portfolio of luciferases is now available with tuned emission wavelengths, kinetics, and high stability. These luciferases can be implemented in the drug discovery and development pipeline, allowing high sensitivity and multiplexing capability.

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Bioluminescence (BL), , the emission of light in living organisms, has become an indispensable tool for a plethora of applications including bioassays, biosensors, and imaging. Current efforts are focused on the obtainment of new luciferases having optimized properties, such as improved thermostability at 37 °C, pH-insensitive emission, high quantum yield, extended kinetics and red-shifted emission. To address these issues we have obtained two new synthetic luciferases, an orange and a red-emitting luciferase, which were designed to achieve high sensitivity (BoLuc) and multiplexing capability (BrLuc) for and biosensing using as a starting template a recently developed thermostable synthetic luciferase (BgLuc).

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The United Nations Agenda 2030 Sustainable Development Goal 6 (SDG 6) aims at ensuring the availability and sustainable management of water and sanitation. The routine monitoring of water contaminants requires accurate and rapid analytical techniques. Laboratory analyses and conventional methods of field sampling still require considerable labor and time with highly trained personnel and transport to a central facility with sophisticated equipment, which renders routine monitoring cumbersome, time-consuming, and costly.

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Adenosine triphosphate (ATP) determination has been used for many decades to assess microbial contamination for hygiene monitoring in different locations and workplace environments. Highly sophisticated methods have been reported, yet commercially available kits rely on a luciferase-luciferin system and require storage and shipping at controlled temperatures (+4 or -20 °C). The applicability of these systems is limited by the need for a secure cold chain, which is not always applicable, especially in remote areas or low-resource settings.

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Biogenic amines (BAs), nitrogenous molecules usually present in different foods, can be considered an indicator of freshness and food quality since their amount increases during food spoilage. Their detection, possibly in real time via the use of smart packaging, is therefore of crucial importance to ensure food safety and to fulfill consumers' demand. To this end, colorimetric sensors are considered one of the most feasible solutions.

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The identification of new strategies to improve the stability of proteins is of utmost importance for a number of applications, from biosensing to biocatalysis. Metal-organic frameworks (MOFs) have been shown as a versatile host platform for the immobilization of proteins, with the potential to protect proteins in harsh conditions. In this work, a new thermostable luciferase mutant has been selected as a bioluminescent protein model to investigate the suitability of MOFs to improve its stability and prompt its applications in real-world applications, for example, ATP detection in portable systems.

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The availability of new bioluminescent proteins with tuned properties, both in terms of emission wavelength, kinetics and protein stability, is highly valuable in the bioanalytical field, with the potential to improve the sensitivity and analytical performance of the currently used methods for ATP detection, whole-cell biosensors, and viability assays among others. We present a new luciferase mutant, called BgLuc, suitable for developing whole-cell biosensors and in vitro biosensors characterized by a bioluminescence maximum of 548 nm, narrow emission bandwidth, favorable kinetic properties, and excellent pH- and thermo-stabilities at 37 and 45 °C and pH from 5.0 to 8.

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One of the major challenges for the modern society, is the development of a sustainable economy also aiming at the valorization of agro-industrial by-products in conjunction with at a significant reduction of generated residues from farm to retail. In this context, the present study demonstrates a biotechnological approach to yield bioactive peptides from a protein fraction obtained as a by-product of the rice starch production. Enzymatic hydrolysis, with the commercial proteases Alcalase and Protamex, were optimized in bioreactor up to 2 L of volume.

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Adenosine-5'-triphosphate (ATP) is the primary energy carrier in all living organisms, and its detection in living cells represents a well-established approach. ATP-driven bioluminescence (BL) relying on the D-luciferin-luciferase reaction is a bioanalytical tool widely employed for monitoring hygiene and microbial contamination of foods.Here, we report a straightforward method for ATP BL detection using an ATP sensing paper fabricated with an alternative freeze-dry procedure.

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The development of predictive sensing tools able to provide rapid information on the different bioactivities of a sample is of pivotal importance, not only to monitor environmental toxicants, but also to understand their mechanisms of action on diverse molecular pathways. This mechanistic understanding is highly important for the characterization of toxicological hazards, and for the risk assessment of chemicals and environmental samples such as surface waters and effluents. Prompted by this need, we developed and optimized a straightforward bioluminescent multiplexed assay which enables the measurement of four bioactivities, selected for their relevance from a toxicological perspective, in bioluminescent microtissues.

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Bioluminescence, that is the emission of light in living organisms, has been extensively explored and applied for diverse bioanalytical applications, spanning from molecular imaging to biosensing. The unprecedented technological evolution of portable light detectors opened new possibilities to implement bioluminescence detection into miniaturized devices. We are witnessing a number of applications, including DNA sequencing, reporter gene assays, DNA amplification for point-of care and point-of need analyses relying on BL.

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Mercury contamination in the environment has reached alarming levels. Due to its persistence and bioaccumulation, mercury is one of the most widespread toxic heavy metals found in air, water and food. Thus, it is mandatory to monitor mercury and its compounds, and the availability of sensitive and rapid biosensors is highly valuable.

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Since the introduction of paper-based analytical devices as potential diagnostic platforms a few decades ago, huge efforts have been made in this field to develop systems suitable for meeting the requirements for the point-of-care (POC) approach. Considerable progress has been achieved in the adaptation of existing analysis methods to a paper-based format, especially considering the chemiluminescent (CL)-immunoassays-based techniques. The implementation of biospecific assays with CL detection and paper-based technology represents an ideal solution for the development of portable analytical devices for on-site applications, since the peculiarities of these features create a unique combination for fitting the POC purposes.

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  • Paper-based lateral-flow immunoassays (LFIAs) are popular in diagnostics, primarily providing qualitative results through visible color changes indicating the presence of specific analytes.
  • Recent advances in immunology, including sensitive tracers and signal amplification, could enhance LFIAs, transforming them into ultrasensitive quantitative tools.
  • The review discusses recent innovations in enzyme-based amplification strategies to improve LFIA performance, highlighting current applications, features, and future challenges in the field.
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The availability of portable analytical devices for on-site monitoring and rapid detection of analytes of forensic, environmental, and clinical interest is vital. We report the development of a portable device for the detection of biochemiluminescence relying on silicon photomultiplier (SiPM) technology, called LuminoSiPM, which includes a 3D printed sample holder that can be adapted for both liquid samples and paper-based biosensing. We performed a comparison of analytical performance in terms of detectability with a benchtop luminometer, a portable cooled charge-coupled device (CCD sensor), and smartphone-integrated complementary metal oxide semiconductor (CMOS) sensors.

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In recent years, there has been an increasing demand for predictive and sensitive in vitro tools for drug discovery. Split complementation assays have the potential to enlarge the arsenal of in vitro tools for compound screening, with most of them relying on well-established reporter gene assays. In particular, ligand-induced complementation of split luciferases is emerging as a suitable approach for monitoring protein-protein interactions.

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Recent advancements in synthetic biology, organic chemistry, and computational models have allowed the application of bioluminescence in several fields, ranging from well established methods for detecting microbial contamination to in vivo imaging to track cancer and stem cells, from cell-based assays to optogenetics. Moreover, thanks to recent technological progress in miniaturized and sensitive light detectors, such as photodiodes and imaging sensors, it is possible to implement laboratory-based assays, such as cell-based and enzymatic assays, into portable analytical devices for point-of-care and on-site applications. This review highlights some recent advances in the development of whole-cell and cell-free bioluminescence biosensors with a glance on current challenges and different strategies that have been used to turn bioassays into biosensors with the required analytical performance.

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The rice-starch processing industry produces large amounts of a protein-rich byproducts during the conversion of broken rice to powder and crystal starch. Given the poor protein solubility, this material is currently discarded or used as animal feed. To fully exploit rice's nutritional properties and reduce this waste, a biotechnological approach was adopted, inducing fermentation with selected microorganisms capable of converting the substrate into peptide fractions with health-related bioactivity.

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The rapid hydrolysis of acetylcholine (ACh), one of the key neurotransmitters in the human body, by the enzyme acetylcholinesterase (AChE) is fundamental for the termination of ACh impulse transmission. Several chemicals, including organophosphorus (OP) pesticides, warfare agents and drugs, are AChE reversible or irreversible inhibitors, thus their rapid on-site detection is of primary importance. Here we report for the first time a chemiluminescence (CL) foldable paper-based biosensor for detection of AChE inhibitors.

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