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Immunosensors are devices that exploit immobilized antibodies to promote the binding of specific analytes related to diseases of medical importance, such as cancer or cardiac dysfunctions. Label-free immunosensors have an important role, due to their simplicity and fast read-out. Here, the proof of concept for an immunosensor based on a 2-D photonic crystal silicon nitride membrane is presented. The device has been fabricated by means of a well-tuned nanofabrication protocol, achieving a high-quality photonic pattern on a large-area membrane (1 mm × 1 mm), and it has been tested for the detection of interleukin-6, getting protein detection at pg/mL concentrations. [ABSTRACT FROM AUTHOR] more...

The development of point-of-care wearable devices capable of measuring insulin concentration has the potential to significantly improve diabetes management and life quality of diabetic patients. However, the lack of a suitable point-of-care device for personal use makes regular insulin level measurements challenging, in stark contrast to glucose monitoring. Herein, we report an electrochemical transdermal biosensor that utilizes a high-density polymeric microneedle array (MNA) to detect insulin in interstitial fluid (ISF). The biosensor consists of gold-coated polymeric MNA modified with an insulin-selective aptamer, which was used for extraction and electrochemical quantification of the insulin in ISF. In vitro testing of biosensor, performed in artificial ISF (aISF), showed high selectivity for insulin with a linear response between 0.01 nM and 4 nM (sensitivity of ∼65 Ω nM -1 ), a range that covers both the physiological and the pathological concentration range. Furthermore, ex vivo extraction and quantification of insulin from mouse skin showed no impact on the biosensor's linear response. As a proof of concept, an MNA-based biosensing platform was utilized for the extraction and quantification of insulin on live mouse skin. In vivo application showed the ability of MNs to reach ISF, extract insulin from ISF, and perform electrochemical measurements sufficient for determining insulin levels in blood and ISF. We believe that our MNA-based biosensing platform based on extraction and quantification of the biomarkers will help move insulin assays from traditional laboratory approaches to personalized point-of-care settings., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.) more...

The development of simple, cost-effective, rapid, and quantitative diagnostic tools remains critical to monitor infectious COVID-19 disease. Although numerous diagnostic platforms, including rapid antigen tests, are developed and used, they suffer from limited accuracy, especially when tested with asymptomatic patients. Here, a unique approach to fabricate a nanochannel-based electrochemical biosensor that can detect the entire virion instead of virus fragments, is demonstrated. The sensing platform has uniform nanoscale channels created by the convective assembly of polystyrene (PS) beads on gold electrodes. The PS beads are then functionalized with bioreceptors while the gold surface is endowed with anti-fouling properties. When added to the biosensor, SARS-CoV-2 virus particles block the nanochannels by specific binding to the bioreceptors. The nanochannel blockage hinders the diffusion of a redox probe; and thus, allows quantification of the viral load by measuring the changes in the oxidation current before and after virus incubation. The biosensor shows a low limit of detection of ≈1.0 viral particle mL -1 with a wide detection range up to 10 8 particles mL -1 in cell culture media. Moreover, the biosensor is able to differentiate saliva samples with SARS-CoV-2 from those without, demonstrating the potential of this technology for translation into a point-of-care biosensor product., (© 2022 The Authors. Small published by Wiley-VCH GmbH.) more...

This article reviews the recent advances in the field of batteryless near-field communication (NFC) sensors for chemical sensing and biosensing. The commercial availability of low-cost commercial NFC integrated circuits (ICs) and their massive integration in smartphones, used as readers and cloud interfaces, have aroused great interest in new batteryless NFC sensors. The fact that coil antennas are not importantly affected by the body compared with other wireless sensors based on far-field communications makes this technology suitable for future wearable point-of-care testing (PoCT) devices. This review first compares energy harvesting based on NFC to other energy-harvesting technologies. Next, some practical recommendations for designing and tuning NFC-based tags are described. Power transfer is key because in most cases, the energy harvested has to be stable for several seconds and not contaminated by undesired signals. For this reason, the effect of the dimensions of the coils and the conductivity on the wireless power transfer is thoroughly discussed. In the last part of the review, the state of the art in NFC-based chemical and biosensors is presented. NFC-based tags (or sensor tags) are mainly based on commercial or custom NFC ICs, which are used to harvest the energy from the RF field generated by the smartphone to power the electronics. Low-consumption colorimeters and potentiostats can be integrated into these NFC tags, opening the door to the integration of chemical sensors and biosensors, which can be harvested and read from a smartphone. The smartphone is also used to upload the acquired information to the cloud to facilitate the internet of medical things (IoMT) paradigm. Finally, several chipless sensors recently proposed in the literature as a low-cost alternative for chemical applications are discussed. more...

Mesoporous thin films are widely used for applications in need of high surface area and efficient mass and charge transport properties. A well-established fabrication process involves the supramolecular assembly of organic molecules (e.g., block copolymers and surfactants) with inorganic materials obtained by sol-gel chemistry. Typically, subsequent calcination in air removes the organic template and reveals the porous inorganic network. A significant challenge for such coatings is the anisotropic shrinkage due to the volume contraction related to solvent evaporation, inorganic condensation, and template removal, affecting the final porosity as well as pore shape, size, arrangement, and accessibility. Here, we show that a two-step calcination process, composed of high-temperature treatment in argon followed by air calcination, is an effective fabrication strategy to reduce film contraction and enhance structural control of mesoporous thin films. Crucially, the formation of a transient carbonaceous scaffold enables the inorganic matrix to fully condense before template removal. The resulting mesoporous films retain a higher porosity as well as bigger pores with extended porous order. Such films present favorable characteristics for mass transport of large molecules. This is demonstrated for lysozyme adsorption into the mesoporous thin films as an example of enzyme storage. more...

This work studies the feasibility of using a battery-less Near-Field Communication (NFC) potentiostat for the next generation of electrochemical point-of-care sensors. A design based on an NFC microchip, a microcontroller, and a custom potentiostat based on an operational amplifier is presented. A proof-of-concept prototype has been designed and used to quantify glucose concentration using commercial glucose test strips from chronoamperometry measurements. The device is harvested and the sensor is read using a mobile phone. The prototype uses an antenna loop covered with ferrite sheets to ensure stable operation of the electronics when the mobile phone is used as reader. The use of ferrite reduces the detuning caused by the proximity of the metal parts of the mobile phone. A comparison with a commercial glucometer device is provided. Results obtained using a commercial glucometer and those provided by the proposed potentiostat show an excellent agreement. more...

Rapid, sensitive, selective and portable virus detection is in high demand globally. However, differentiating non-infectious viral particles from intact/infectious viruses is still a rarely satisfied sensing requirement. Using the negative space within monolayers of polystyrene (PS) spheres deposited directly on gold electrodes, we fabricated tuneable nanochannels decorated with target-selective bioreceptors that facilitate the size-selective detection of intact viruses. Detection occurred through selective nanochannel blockage of diffusion of a redox probe, [Fe(CN) 6 ] 3/4- , allowing a quantifiable change in the oxidation current before and after analyte binding to the bioreceptor immobilised on the spheres. Our model system involved partial surface passivation of the mono-assembled PS spheres, by silica glancing angle deposition, to confine bioreceptor immobilisation specifically to the channels and improve particle detection sensitivity. Virus detection was first optimised and modelled with biotinylated gold nanoparticles, recognised by streptavidin immobilised on the PS layer, reaching a low limit of detection of 37 particles/mL. Intact, label-free virus detection was demonstrated using MS2 bacteriophage (~23-28 nm), a marker of microbiological contamination, showing an excellent limit of detection of ~1.0 pfu/mL. Tuneable nanochannel geometries constructed directly on sensing electrodes offer label-free, sensitive, and cost-efficient point-of-care biosensing platforms that could be applied for a wide range of viruses. more...

The biomolecular contents of extracellular vesicles, such as exosomes, have been shown to be crucial in intercellular communication and disease propagation. As a result, there has been a recent surge in the exploration of novel biosensing platforms that can sensitively and specifically detect exosomal content such as proteins and nucleic acids, with a view toward application in diagnostic assays. Here, we demonstrate dual-mode and label-free detection of plasma exosomes using an electrochemical quartz crystal microbalance with dissipation monitoring (EQCM-D). The platform adopts a direct immunosensing approach to effectively capture exosomes via their surface protein expression of CD63. By combining QCM-D with a tandem in situ electrochemical impedance spectroscopy measurement, we are able to demonstrate relationships between mass, viscoelasticity and impedance inducing properties of each functional layer and analyte. In addition to lowering the limit of detection (by a factor of 2-4) to 6.71 × 10 7 exosome-sized particles (ESP) per mL in 25% v/v serum, the synergy between dissipation and impedance response introduces improved sensing specificity by offering further distinction between soft and rigid analytes, thereby promoting EQCM-D as an important technique for exosome analysis. more...

Wearable technologies have great potential in health monitoring and disease diagnostics. As a consequence, interest in the study of wearable sensors has dramatically increased over recent years. Successful translation of this technology from research prototypes to commercial products requires addressing some of the major challenges faced by wearable sensors such as loss of, and damage in, the biological recognition layer of the skin-interfaced sensors. In this work, we propose a solution to this challenge by integrating micropillar array (MPA) surfaces as part of the sensing layer with the aim to protect and prevent the loss of the enzyme layer from mechanical stress while the sensor is worn. The proposed wearable sensing patch is composed of reference, counter, and working electrodes, all made of MPAs and is designed for measuring glucose in sweat. MPA sensing patch has a wide linear range of 50 μM to 1.4 mM, a sensitivity of 4.7 ± 0.8 μA mM -1 , and a limit of detection of 26 ± 5 μM. The glucose sensing patch was tested using human sweat where glucose-level changes were successfully measured before and after meal consumption. The developed patch provides an alternative solution to the problem of the damage to the sensor microenvironment upon wear. But in addition, it also offers a user-friendly, cost-effective, and reliable sweat analysis platform with significant potential in health monitoring applications. more...

Electrochemical devices for transdermal monitoring of key biomarkers are the potential next frontier of wearable technologies for point-of-care disease diagnosis, including Cancer in which Cancer is the leading cause of death worldwide with estimated 10 million deaths in 2018 according to the World Health Organization and breast cancer is one of the five most common causes of cancer death with over two million cases recorded in 2018. Early diagnosis and prognosis based on monitoring of breast cancer biomarkers is of high importance. In this work, high-density gold coated silicon microneedle arrays (Au-Si-MNA) were simultaneously used as biomarker extraction platform and electrochemical transducer, enabling the selective immunocapture of epidermal growth factor receptor 2 (ErbB2), a key breast cancer biomarker, and its subsequent quantification. The analytical performance of the device was tested in artificial interstitial fluid exhibiting a linear response over a wide concentration range from 10 to 250 ng/mL, with a detection limit of 4.8 ng/mL below the biomarker levels expected in breast cancer patients. As a proof of concept, the immunosensor demonstrated its ability to successfully extract ErbB2 from a phantom gel mimicking the epidermis and dermis layers, and subsequently quantify it showing a linear range from 50 to 250 ng/mL and a detection limit of 25 ng/mL. The uniqueness of this sensing platform combining direct transdermal biomarker extraction and quantification opens up new avenues towards the development of high performing wearable point-of-care devices., (Copyright © 2021 Elsevier B.V. All rights reserved.) more...

Porous polymeric membranes have shown great potential in biological and biomedical applications such as tissue engineering, bioseparation, and biosensing, due to their structural flexibility, versatile surface chemistry, and biocompatibility. This review outlines the advantages and limitations of the fabrication techniques commonly used to produce porous polymeric membranes, with especial focus on those featuring nano/submicron scale pores, which include track etching, nanoimprinting, block-copolymer self-assembly, and electrospinning. Recent advances in membrane technology have been key to facilitate precise control of pore size, shape, density and surface properties. The review provides a critical overview of the main biological and biomedical applications of these porous polymeric membranes, especially focusing on drug delivery, tissue engineering, biosensing, and bioseparation. The effect of the membrane material and pore morphology on the role of the membranes for each specific application as well as the specific fabrication challenges, and future prospects of these membranes are thoroughly discussed. more...

Nanotechnology's rapid development of nanostructured materials with disruptive material properties has inspired research for their use as electrocatalysts to potentially substitute enzymes. Herein, a novel electrocatalytic nanomaterial was constructed by growing gold nanograss (AuNG) on 2D nanoassemblies of gold nanocubes (AuNC). The resulting structure (NG@NC) was used for the detection of H 2 O 2 via its electrochemical reduction. The NG@NC electrode displayed a large active surface area, resulting in improved electron transfer efficiency. On the nanoscale, AuNG maintained its structure, providing high stability and reproducibility of the sensing platform. Our nanostructured electrode showed excellent catalytic activity towards H 2 O 2 at an applied potential of -0.5 V vs Ag/AgCl. This facilitated H 2 O 2 detection with excellent selectivity in an environment like human urine, and a linear response from 50 µM to 30 mM, with a sensitivity of 100.66 ± 4.0 μA mM -1  cm -2 . The NG@NC-based sensor hence shows great potential in nonenzymatic electrochemical sensing., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.) more...

A label-free electrochemical detection platform for the sensitive and rapid detection of Flightless I (Flii) protein, a biomarker of wound chronicity, has been developed using nanoporous anodic alumina (NAA) membranes modified with Flii antibody recognition sites. The electrochemical detection is based on the nanochannel blockage experienced upon Flii capture by immobilized antibodies within the nanochannels. This capture impedes the diffusion of redox species [[Fe(CN) 6 ] 4-/3- ] toward a gold electrode attached at the backside of the modified NAA membrane. Partial blockage causes a decrease in the oxidation current of the redox species at the electrode surface which is used as an analytical signal by the reported biosensor. The resulting biosensing system allows detection of Flii at the levels found in wounds. Two types of assays were tested, sandwich and direct, showing <3 and 2 h analysis time, respectively, a significant reduction in time from the nearly 48 h required for the conventional Western blot assay. Slightly higher sensitivity values were observed for the sandwich-based strategy. With faster analysis, lack of matrix effects, robustness, ease of use and cost-effectiveness, the developed sensing platform has the potential to be translated into a point-of-care (POC) device for chronic wound management and as a simple alternative characterization tool in Flii research., (Copyright © 2020 Rajeev, Melville, Cowin, Prieto-Simon and Voelcker.) more...

We report the first method to introduce differential functionalities in the interior pore walls and exterior surface of highly stable thermally hydrocarbonised porous silicon (THCpSi) films. The approach exploits the hydrophobicity of the hydrosilylated THCpSi to, first, selectively functionalise the external surface, and subsequently derivatise the hydrophobic internal pore walls. more...

Lipopolysaccharides (LPS) are the major component of the outer membrane of all Gram-negative bacteria and some cyanobacteria and are released during growth and cell death. LPS pose a potential health risk in water, causing acute respiratory illnesses, inhalation fever, and gastrointestinal disorders. The need for rapid and accurate detection of LPS has become a major priority to facilitate more timely and efficacious intervention and, hence, avoid unsafe water distribution. In this context, a porous silicon membrane (pSiM)-based electrochemical biosensor was developed for direct and sensitive detection of LPS. pSiM, featuring arrays of nanochannels, was modified with polymyxin B (PmB), an antimicrobial peptide with strong affinity to LPS. Detection of LPS was based on measuring the changes in the diffusion through the nanochannels of an electroactive species added in solution, caused by the nanochannel blockage upon LPS binding to PmB. Results showed a limit of detection of 1.8 ng/mL, and a linear response up to 10,000 ng/mL spiked in buffer. Selectivity of the sensor toward potential interfering species in water supplies was also assessed. Sensor performance was then evaluated in water samples from a water treatment plant (WTP), and detection of LPS well below the levels encountered in episodes of water contamination and in humidifiers was demonstrated. The same platform was also tested for bacterial detection including Pseudomonas aeruginosa and Escherichia coli spiked in water samples from a WTP. Considering its performance characteristics, this platform represents a promising screening tool to identify the presence of LPS in water supplies and provide early warning of contamination events. more...

A novel pore blockage-based electrochemical immunosensor based on the combination of 100 nm-magnetic nanoparticles (MNPs), as signal enhancers, and 200 nm-pore diameter nanoporous anodic alumina (NAA) membranes, as sensing platform, is reported. A peptide conjugate mimicking flightless I (Flii), a wound healing biomarker, was chosen as target analyte. The sensing platform consists of an anti-Flii antibody (Ab1)-modified NAA membrane attached onto a gold electrode. Anti-KLH antibody (Ab2)-modified MNPs (MNP-Ab2) were used to selectively capture the Flii peptide conjugate in solution. Sensing was based on pore blockage of the Ab1-modified NAA membrane caused upon specific binding of the MNP-Ab2-analyte complex. The degree of pore blockage, and thus the concentration of the Flii peptide conjugate in the sample, was measured as a reduction in the oxidation current of a redox species ([Fe(CN) 6 ] 4- ) added in solution. We demonstrated that pore blockage is drastically enhanced by applying an external magnetic field at the membrane backside to facilitate access of the MNP-Ab2-analyte complex into the pores, and thus ensure its availability to bind to the Ab1-modified NAA membrane. Combining the pore blockage-based electrochemical magnetoimmunosensor with an externally applied magnetic field, a limit of detection (LOD) of 0.5 ng/ml of Flii peptide conjugate was achieved, while sensing in the absence of magnetic field could only attain a LOD of 1.2 μg/ml. The developed sensing strategy is envisaged as a powerful solution for the ultra-sensitive detection of an analyte of interest present in a complex matrix. more...

The development of enzyme-responsive hyaluronic acid methacrylate (HYAMA)-coated porous silicon (pSi) films and their application in electrochemical diagnostic devices for the in situ detection of the enzyme hyaluronidase (hyal), which is secreted by Staphylococcus aureus (S. aureus) bacteria, are reported. The approach relies on a HYAMA-pSi electrode made of thermally hydrocarbonized pSi (pSi-THC) that is impregnated with crosslinked HYAMA/polyethylene glycol diacrylate (PEGDA) hydrogels. The enzymatic degradation of HYAMA by bacterial hyal is monitored by differential pulse voltammetry (DPV) utilizing pSi-THC as a working electrode and ferro/ferricyanide (FF) as external redox probe. The degradation of HYAMA results in reduced diffusion of the redox probe through the partially charged film, thereby enabling the detection of hyal by DPV. In addition to the determination of the concentration-dependent response in NaOAc buffer (pH 5.2), the detection of hyal as indicator for the presence of S. aureus bacteria above a threshold level in bacterial supernatants and artificial wound fluid is highlighted., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.) more...

There is a strong and growing demand for compact, portable, rapid, and low-cost devices to detect biomarkers of interest in clinical and point-of-care diagnostics. Such devices aid in early diagnosis of diseases without the need to rely on expensive and time-consuming large instruments in dedicated laboratories. Over the last decade, numerous biosensors have been developed for detection of a wide range of clinical biomarkers including proteins, nucleic acids, growth factors, and bacterial enzymes. Various transduction techniques have been reported based on biosensor technology that deliver substantial advances in analytical performance, including sensitivity, reproducibility, selectivity, and speed for monitoring a wide range of human health conditions. Nanoporous anodic alumina (NAA) has been used extensively for biosensing applications due to its inherent optical and electrochemical properties, ease of fabrication, large surface area, tunable properties, and high stability in aqueous environment. This review focuses on NAA-based biosensing systems for detection of clinically significant biomarkers using various detection techniques with the main focus being on electrochemical and optical transduction methods. The review covers an overview of the importance of biosensors for biomarkers detection, general (surface and structural) properties and fabrication of NAA, and NAA-based biomarker sensing systems., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.) more...

The emergence of nanostructured materials has opened new horizons in the development of next generation biosensors. Being able to control the design of the electrode interface at the nanoscale combined with the intrinsic characteristics of the nanomaterials engenders novel biosensing platforms with improved capabilities. The purpose of this review is to provide a comprehensive and critical overview of the latest trends in emerging nanostructured electrochemical biosensors. A detailed description and discussion of recent approaches to construct label-free electrochemical nanostructured electrodes is given with special focus on pathogen detection for environmental monitoring and food safety. This includes the use of nanoscale materials such as nanotubes, nanowires, nanoparticles, and nanosheets as well as porous nanostructured materials including nanoporous anodic alumina, mesoporous silica, porous silicon, and polystyrene nanochannels. These platforms may pave the way toward the development of point-of-care portable electronic devices for applications ranging from environmental analysis to biomedical diagnostics. more...

We report on the design and testing of photonic biosensors for the type-selective detection of different types of matrix metalloproteinases (MMPs). The ability to detect a panel of different MMP types has important implications for prognosis of wound healing. We combine the immunocapture of MMPs on dispersed magnetic nanoparticles modified with antibodies specific for target MMPs (immuno-magNPs) with subsequent MMP detection upon fluorogenic peptide cleavage in porous silicon resonant microcavity (pSiRM) architectures. We report fast, sensitive, and type-selective detection of MMPs directly in wound fluid. This study sets the scene for downstream developments of multiparametric biosensors as point-of-care (POC) prognostic tools that may step-change chronic wound management. more...

Bacterial wound infections can cause septicemia and lead to limb amputation or death. Therefore, early detection of bacteria is important in chronic wound management. Here, an optical biosensor based on porous silicon resonant microcavity (pSiRM) structure modified with fluorogenic peptide substrate is demonstrated to detect the presence of Sortase A (SrtA), a bacterial enzyme found in the cell membrane protein of Staphylococcus aureus . The combination of fluorescence enhancement effects of the pSiRM architecture with the incorporation of SrtA fluorogenic peptide substrate within the pSi matrix enables the sensing of SrtA with an outstanding limit of detection of 8 × 10 -14 m. Modification of the pSiRM structure with microscale spots of two fluorogenic peptide substrates, one specific for SrtA and the other for matrix metalloproteinases, effectively demonstrates the feasibility to perform multiplexed biomarker analysis. The results in this study highlight the potential of the pSiRM sensing platform as a point-of-care diagnostic tool for biomarkers of bacterial wound infection. more...

The up-regulation of L-lactate dehydrogenase (LDH), an intracellular enzyme present in most of all body tissues, is indicative of several pathological conditions and cellular death. Herein, we demonstrate LDH detection using porous silicon (pSi) microcavities as a luminescence-enhancing optical biosensing platform. Non-fluorescent resazurin was covalently attached onto the pSi surface via thermal hydrocarbonisation, thermal hydrosylilation and acylation. Each surface modification step was confirmed by means of FTIR and the optical shifts of the resonance wavelength of the microcavity. Thermal hydrocarbonisation also afforded excellent surface stability, ensuring that the resazurin was not reduced on the pSi surface. Using a pSi microcavity biosensor, the fluorescence signal upon detection of LDH was amplified by 10 and 5-fold compared to that of a single layer and a detuned microcavity, respectively, giving a limit of detection of 0.08 U/ml. The biosensor showed a linear response between 0.16 and 6.5 U/ml, covering the concentration range of LDH in normal as well as damaged tissues. The biosensor was selective for LDH and did not produce a signal upon incubation with another NAD-dependant enzyme L-glutamic dehydrogenase. The use of the pSi microcavity as a sensing platform reduced reagent usage by 30% and analysis time threefold compared to the standard LDH assay in solution., (Copyright © 2015 Elsevier B.V. All rights reserved.) more...