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A sensitive and robust electrochemical cholinesterase-based sensor was developed to detect the quaternary ammonium (QAs) biocides most frequently found in agri-food industry wash waters: benzalkonium chloride (BAC) and didecyldimethylammonium chloride (DDAC). To reach the maximum residue limit of 28 nM imposed by the European Union (EU), two types of cholinesterases were tested, acetylcholinesterase (AChE, from Drosophila melanogaster) and butyrylcholinesterase (BChE, from horse serum). The sensors were designed by entrapping AChE or BChE on cobalt phthalocyanine-modified screen-printed carbon electrodes. The limits of detection (LOD) of the resulting biosensors were 38 nM for DDAC and 320 nM for BAC, using, respectively, AChE and BChE. A simple solid-phase extraction step was used to concentrate the samples before biosensor analysis, allowing for the accurate determination of DDAC and BAC in tap water with limits of quantification (LOQ) as low as 2.7 nM and 5.3 nM, respectively. Additional assays demonstrated that the use of a phosphotriesterase enzyme allows for the total removal of interferences due to the possible presence of organophosphate insecticides in the sample. The developed biosensors were shown to be stable during 3 months storage at 4 °C. [ABSTRACT FROM AUTHOR]

This paper focuses on the development of a novel electrode based on boron-doped diamond nanosheet full-volume-enriched screen-printed carbon electrodes (BDDPE) for use as an impedimetric biosensor. Impedimetric biosensors offer high sensitivity and selectivity for virus detection, but their use as point-of-care devices is limited by the complexity of nanomaterials' architecture and the receptor immobilisation procedures. The study presents a two-step modification process involving the electroreduction of diazonium salt at the BDDPE and the immobilisation of antibodies using zero-length cross-linkers for a selective impedimetric biosensor of Haemophilus influenzae (Hi). The incorporation of diamond nanosheets into BDDPE leads to enhanced charge transfer and electrochemical behaviour, demonstrating greatly improved electrochemically active surface area compared with unmodified screen-printed electrodes (by 44% and 10% on average for [Ru(NH3)6]Cl2 and K3[Fe(CN)6], respectively). The presented sensing system shows high specificity towards protein D in Hi bacteria, as confirmed by negative controls against potential interference from other pathogens, with an estimated tolerance limit for interference under 12%. The Hi limit of detection by electrochemical impedance spectroscopy was 1 CFU/mL (measured at − 0.13 V vs BDDPE pseudo-reference), which was achieved in under 10 min, including 5 min sample incubation in the presence of the analyte. [ABSTRACT FROM AUTHOR]

We introduce a bifunctional architecture to construct biosensors on solution-blow spinning fiber mats of polylactic acid (PLA) and polyethylene glycol (PEG). PLA/PEG mats acted as substrate for printing electrodes and as matrix to immobilize glucose oxidase (GOx). Prussian Blue nanoparticles (PB) decorated working electrodes to detect H2O2 from enzymatic catalysis at a low applied potential (0 V vs. Ag/AgCl) with detection limit of 0.197 mM. The inexpensive bifunctional device (

We report on hydrogen peroxide sensors made through a single printing step with carbon ink containing catalytically synthesized Prussian blue nanoparticles. Despite their reduced sensitivity, the resulting bulk-modified sensors displayed both a wider linear calibration range (5 × 10−7–1 × 10−3 M) and an approximately four times lower detection limit versus the surface-modified sensors due to the dramatically decreased noise resulting in, on average, a six times higher signal-to-noise ratio. The corresponding glucose and lactate biosensors demonstrated similar and even higher sensitivities compared to those of biosensors based on surface-modified transducers. The biosensors have been validated through analysis of human serum. The decreased time and cost for production of single printing step bulk-modified transducers, as well as their analytical performance characteristics, which are advantageous over conventional surface-modified ones, would be expected to enable their wide use in (bio)sensorics. [ABSTRACT FROM AUTHOR]

Human Papilloma Virus 16 (HPV 16) is the well-known causative species responsible for triggering cervical cancer. When left undiagnosed and untreated, this disease leads to life-threatening events among the female populace, especially in developing nations where healthcare resources are already being stretched to their limits. Considering various drawbacks of conventional techniques for diagnosing this highly malignant cancer, it becomes imperative to develop miniaturized biosensing platforms which can aid in early detection of cervical cancer for enhanced patient outcomes. The current study reports on the development of an electrochemical biosensor based on reduced graphene oxide (rGO)/DNA hybrid modified flexible carbon screen-printed electrode (CSPE) for the detection of HPV 16. The carbon-coated SPEs were initially coated with rGO followed by probe DNA (PDNA) immobilization. The nanostructure characterization was performed using UV-Vis spectroscopy, Fourier transform infrared (FTIR) spectroscopy, Raman spectroscopy and X-ray diffraction (XRD) techniques. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were employed to study the electrochemical characterization of the nano-biohybrid sensor surface. The optimization studies and analytical performance were assessed using differential pulse voltammetry (DPV), eventually exhibiting a limit of detection (LoD) ~2 pM. The developed sensor was found to be selective solely to HPV 16 target DNA and exhibited a shelf life of 1 month. The performance of the developed flexible sensor further exhibited a promising response in spiked serum samples, which validates its application in future point-of-care scenarios. [ABSTRACT FROM AUTHOR]

Screen-printed electrodes-based sensors can be successfully used to determine all kinds of analytes with great precision and specificity. However, obtaining a high-quality sensor can be difficult due to factors such as lack of reproducibility, surface contamination or other manufacturing challenges. An important step in ensuring reproducible results is the cleaning step. The aim of the current work is to help researchers around the world who struggle with finding the most suitable method for cleaning screen-printed electrodes. We evaluated the cleaning efficiency of different chemical compounds and cleaning methods using cyclic voltammetry and electrochemical impedance spectroscopy. The percentage differences in polarization resistance (Rp) before and after cleaning were as follows: acetone—35.33% for gold and 49.94 for platinum; ethanol—44.50% for gold and 81.68% for platinum; H2O2—47.34% for gold and 92.78% for platinum; electrochemical method—3.70% for gold and 67.96% for platinum. Thus, we concluded that all the evaluated cleaning methods seem to improve the surface of both gold and platinum electrodes; however, the most important reduction in the polarization resistance (Rp) was obtained after treating them with a solution of H2O2 and multiple CV cycles with a low scanning speed (10 mV/s). [ABSTRACT FROM AUTHOR]

As one of the three major chronic diseases, diabetes often causes many complications, which can affect various parts of the body and even threaten the life of the patients. At present, the situation of diabetes in the world is quite serious. Accurate detection of blood glucose is very important for the diagnosis, treatment and medication of diabetes as well as the self-management of diabetic patients. In this paper, an electrochemical glucose biosensor was developed based on screen-printed electrode (SPE) modified with composite material of graphene aerogel (GA) and Prussian blue (PB) (denoted as GA@PB), which was fabricated via chemical reduction using L-ascorbic acid as a reducing agent through a freeze-drying process. Glucose was specifically captured by glucose oxidase (GOx) which were immobilized into the GA@PB by chitosan. The structure and performance of the sensor were characterized by scanning electron microscopy (SEM), Raman spectroscopy measurements, Fourier transform infrared spectrometer (FTIR), cyclic voltammetry (CV) and amperometric detection. The sensor exhibited a linear range of 0.5–6.0 mmol·L−1 with limit of detection (LOD) of 0.15 mmol·L−1, indicating that the combination of graphene aerogel and Prussian blue possess well conductivity and catalytic performance. [ABSTRACT FROM AUTHOR]

We report on hydrogen peroxide sensors made through a single printing step with carbon ink containing catalytically synthesized Prussian blue nanoparticles. Despite their reduced sensitivity, the resulting bulk-modified sensors displayed both a wider linear calibration range (5 × 10−7–1 × 10−3 M) and an approximately four times lower detection limit versus the surface-modified sensors due to the dramatically decreased noise resulting in, on average, a six times higher signal-to-noise ratio. The corresponding glucose and lactate biosensors demonstrated similar and even higher sensitivities compared to those of biosensors based on surface-modified transducers. The biosensors have been validated through analysis of human serum. The decreased time and cost for production of single printing step bulk-modified transducers, as well as their analytical performance characteristics, which are advantageous over conventional surface-modified ones, would be expected to enable their wide use in (bio)sensorics.

The indiscriminate use of herbicides in agriculture contributes to soil and water pollution, with important endangering consequences on the ecosystems. Among the available analytical systems, algal biosensors have demonstrated to be valid tools thanks to their high sensitivity, cost-effectiveness, and eco-design. Herein, we report the development of a dual electro-optical biosensor for herbicide monitoring, based on Chlamydomonas reinhardtii whole cells immobilised on paper-based screen-printed electrodes modified with carbon black nanomaterials. To this aim, a systematic study was performed for the selection and characterisation of a collection among 28 different genetic variants of the alga with difference response behaviour towards diverse herbicide classes. Thus, CC125 strain was exploited as case study for the study of the analytical parameters. The biosensor was tested in standard solutions and real samples, providing high sensitivity (detection limit in the pico/nanomolar), high repeatability (RSD of 5% with n = 100), long lasting working (10 h) and storage stability (3 weeks), any interference in the presence of heavy metals and insecticides, and low matrix effect in drinking water and moderate effect in surface one. [ABSTRACT FROM AUTHOR]

A highly sensitive sensor for quantification of uric acid (UA) directly in body fluids (saliva and sweat) is reported, working at a potential as low as 0.0 V vs Ag/AgCl. New mixed hydroxide materials exhibiting stable electrocatalytic responses from alkaline to acidic media were prepared, their structure was thoroughly characterized, and the electrochemical properties of the modified FTO (fluorine-doped tin oxide) electrodes were evaluated for UA determination by cyclic voltammetry, chronoamperometry, and batch injection analysis. A very low limit of detection (2.3 × 10−8 mol L−1) with good repeatability (RSD = 3.2% for 30 successive analyses) was achieved based on a fast and simple BIA procedure. Finally, α-Ni0.75Zn0.25(OH)2 screen-printed electrodes (SPE) were developed for the measurement of UA directly in real saliva and sweat samples, without interference of ascorbic acid, acetaminophen, lactate, and glucose at their typical concentrations present in those body fluids, revealing high potential for application as disposable sensors in biological systems. [ABSTRACT FROM AUTHOR]

[Display omitted] • The AuNPs/rGO nanocomposites exhibit excellent electrochemical properties. • A ssDNA-hDNA coupling biosensor with strong "on–off" signal has been developed. • This cost-effective biosensor is reusable upon re-hatching of signal probes. The DCL gene in Fusarium oxysporum f. sp. cubense Race 4 (Foc4) is a pivotal pathogenic factor causing banana fusarium wilt. Precise DCL detection is crucial for Foc4 containment. Here, we present a novel ssDNA-hDNA coupling electrochemical biosensor for highly specific DCL detection. The sensing interface was formed via electrodeposition of a composite containing reduced graphene oxide (rGO) and gold nanoparticles (AuNPs) onto a carbon screen-printed electrode (SPE), followed by thiol-modified ssDNA functionalization. Additionally, the incorporation of hDNA, with methylene blue (MB) at both ends, binds to ssDNA through base complementarity, forming an ssDNA-hDNA coupling probe with bismethylene blue. This sensing strategy relies on DCL recognition by the hDNA probe, leading to DNA hairpin unfolding and detachment of hDNA bearing two MBs from ssDNA, generating a robust "on-off" signal. Empirical results demonstrate the sensor's amplified electrical signals, reduced background currents, and an extended detection range (6.02 × 106–3.01 × 1010 copies/μL) with a limit of detection (3.01 × 106 copies/μL) for DCL identification. We applied this sensor to analyze soil, banana leaves, and fruit samples, confirming its high specificity and stability. Moreover, post-sample detection, the sensor exhibits reusability, offering a cost-effective and rapid approach for banana wilt detection. [ABSTRACT FROM AUTHOR]

Nickel is naturally present in drinking water and many dietary items, which expose the general population to nickel ingestion. This heavy metal can have a variety of harmful health effects, causing allergies and skin disorders (i.e., dermatitis), lung, cardiovascular, and kidney diseases, and even certain cancers; therefore, nickel detection is important for public health. Recent innovations in the development of biosensors have demonstrated they offer a powerful new approach over conventional analytical techniques for the identification and quantification of user-defined compounds, including heavy metals such as nickel. We optimized five candidate nickel-biosensing receptors, and tested each for efficiency of binding to immobilization elements on screen-printed electrodes (SPEs). We characterized the application of nickel-detecting biosensors with four different cultivated vegetables. We analyzed the efficiency of each nickel-detecting biosensor by potentiostat and atomic absorption spectrometry and compared the results from the sample analytes. We then analyzed the performance characteristics and responses of assembled biosensors, and show they are very effective at measuring nickel ions in food, especially with the urease-alginate biosensor affixed to silver SPEs, measured by cyclic voltammetry (sensitivity—2.1921 µA Mm−1 cm−2 and LOD—0.005 mg/L). Given the many advantages of biosensors, we describe an optimization pipeline approach to the application of different nickel-binding biosensors for public health, nutrition, and consumer safety, which are very promising.

• We present the production of an electrochromic screen-printable paste based on Prussian Blue. • These electrodes display high reversibility and electrocatalysis toward H 2 O 2 reduction. • These blue electrodes enable both the electrochemical and optical quantitative determination of hydrogen peroxide. • The reported blue electrodes are highly suitable transducers for the construction of oxidase-based biosensors. • A glucose biosensor is presented to demonstrate the spectroelectrochemical capabilities of these new electrodes. Prussian Blue (PB)-modified graphite screen-printed electrodes are increasingly being used in electrochemical biosensors. However, they do not allow the observation of the electrochromism of PB. This work presents the construction of PB-based, electrochromic screen-printed biosensors. Although electrically more resistive than their graphite counterparts, these new PB-based electrodes enable both the amperometric and colorimetric detection of hydrogen peroxide. This is the first time that this has been achieved using screen-printed electrodes, and we demonstrate it spectroelectrochemically on a glucose biosensor. The biosensor electrochemical performance equals that of previously reported PB/graphite electrodes, being able to detect down to 4 μM H 2 O 2 and 54 μM glucose. At the same time, and in contrast to PB/graphite electrodes, the new PB-based electrodes afford the optical detection of these two analytes down to 1.2 μM and 15 μM, respectively. The dynamic ranges of the glucose biosensors obtained at the PB-based electrodes are 0.1–1 mM (amperometric) and 0.025–2.5 mM (Colorimetric), matching the physiological glucose concentration range in body fluids other than blood or serum. [ABSTRACT FROM AUTHOR]

The characteristics of biosensors based on screen-printed electrodes containing glucose oxidase, alcohol oxidase, lactate oxidase, and a mixture of glucose oxidase and γ-amylase are studied. The enzymes have been immobilized in a conductive hydrogel of cross-linked bovine serum albumin modified with carbon nanotubes. The developed biosensors are applicable for the quantification of glucose, ethanol, lactic acid, and starch in fermentation media. The analytical and metrological characteristics of these biosensors are determined. The developed biosensor system allows for quantification of glucose, ethanol, lactate, and starch in the concentration range of 0.1–2.7, 1–27, 0.02–73 mM, and 0.6–21 g/L, respectively. Samples of alcoholic and fermented milk products, kvass, and fermentation mass have been analyzed. Statistical analysis of the assay results shows that glucose, ethanol, lactic acid, and starch concentrations determined by biosensors insignificantly differ from those determined by the reference methods. The developed set of biosensors based on specifically selected enzymes can be used for effective monitoring of biotechnological processes as an alternative to the conventional analytical methods. [ABSTRACT FROM AUTHOR]

Abstract A small, standalone electrochemical hybrid sensor platform, combining flexible electronics and screen-printed electrodes, is demonstrated in the determination of capsaicin through adsorptive stripping voltammetry. The sensing scheme was simplified to be compatible with a low-cost device. The simplification involved eliminating the need for additional modification of the electrode and employing a coarsely stepped squarewave voltammetry, a technique which is applicable with less sophisticated instrumentation. This architecture was found to be suitable for concentrations up to at least 5000 µM with a detection limit of 1.98 µM. The screen-printed carbon graphite electrodes were made reusable through an ethanol rinsing protocol. The effect of ethanol/buffer volumetric ratio in the test sample was shown to greatly influence the analytical data, and a fixed 10% (v/v) was chosen as a compromise between signal-to-noise ratio and not exceeding the solubility limit of the desired upper range. Graphical abstract fx1 Highlights • Capsaicin detection demonstrated by coarsely stepped cyclic squarewave voltammetry. • Scheme implemented on electrochemical sensor platform with integrated electrodes. • Unmodified electrodes able to detect highly concentrated capsaicin. • Electrodes made reusable through ethanol cleaning protocol. • Ethanol content in sample solution found to greatly influence the analytical signals. [ABSTRACT FROM AUTHOR]

A sensitive electrochemical detection method for Salmonella spp. was described, based on the use of Hechtia argentea lectin immobilised on a screen-printed gold electrode. The lectin was extracted from Hechtia argentea, a plant belonging to the Bromeliaceae family. The lectin with molecular weight near 27.4 kDa showed selectivity towards D-mannose, contained on the lipopolysaccharide cell wall of Salmonella spp. Carbohydrate selectivity of the lectin was measured as a change in impedance with respect to concentration. The binding of the bacteria to the biosensor surface increased impedance with increasing concentrations of Salmonella spp., achieving a linear range of detection of 15–2.57 × 107 CFU mL−1, with a limit of detection of 5 CFU mL−1. Increases in impedance were measured using electrochemical impedance spectroscopy and analysed using Nyquist plots. The biosensor was applied in analysis of hen egg samples, and the results were consistent with those obtained using the official analysis methodology.

Due to rapidly spreading infectious diseases and the high incidence of other diseases such as cancer or metabolic syndrome, there is a continuous need for the development of rapid and accurate diagnosis methods. Screen-printed electrodes-based biosensors have been reported to offer reliable results, with high sensitivity and selectivity and, in some cases, low detection limits. There are a series of materials (carbon, gold, platinum, etc.) used for the manufacturing of working electrodes. Each version comes with advantages, as well as challenges for their functionalization. Thus, the aim is to review the most promising biosensors developed using screen-printed electrodes for the detection/quantification of proteins, biomarkers, or pathogenic microorganisms.

Abstract Herein, we report an impedimetric DNA-based aptamer sensor for a single-step detection of B. anthracis spore simulant (B. cereus spore). Specifically, we designed a miniaturized label-free aptasensor for B. cereus spores based on a gold screen-printed electrode functionalized with B. cereus spores-binding aptamer (BAS-6R). Several parameters were optimized to fabricate the aptasensor such as the concentration of DNA aptamer solution (0.5 µM), the time (48 h), the temperature (4 °C), and the pH (7.5) for aptamer immobilization on the working electrode surface. Once the aptasensor was developed, it was tested against B. cereus spores 14579 evaluating the effect of incubation time and MgCl 2 concentration. Under the optimized conditions (incubation time equal to 3 h and absence of MgCl 2), B. cereus spores 14579 were detected with a linear range between 104 CFU/ml and 5 × 106 CFU/ml and a detection limit of 3 × 103 CFU/ml. Furthermore, the study of selectivity toward B. cereus 11778, B. subtilis , Legionella pneumophila , and Salmonella Typhimurium has demonstrated the capability of this sensor to detect B. cereus spores, proving the suitability of the DNA-based sensing element combined with a portable instrument for a label-free measurement on site of B. anthracis spore simulant. Highlights • Miniaturized aptasensor for Bacillus anthracis spore detection. • Label free detection exploiting electrochemical impedance spectroscopy. • Aptamer immobilised onto gold-based screen printed electrodes. • Measurement carried out with portable potentiostat. [ABSTRACT FROM AUTHOR]

Abstract In this work it is explored a real applicability of miniaturised and portable biosensing technology for the estimation of total phenolic content in 15 different commercial beers by applying direct amperometry. Gold nanoparticles screen-printed electrodes were thoroughly modified with tyrosinase (Tyr-AuNPS-SPCEs), which was immobilised on the surface by crosslinking with glutaraldehyde. All chemical and instrumental variables involved in the electrochemical method were optimised to develop a reliable and powerful tool to estimate rapidly the content of phenolic compounds in complex beer samples. Catechol, phenol, caffeic acid and tyrosol were analysed individually using the proposed methodology and good analytical and kinetic performances were obtained. Total phenolic content in tested beers (high fermented, low fermented and non-alcoholic) were expressed as mg L−1 of tyrosol, which is one of the major phenolic compound reported in beer. Moreover, the developed amperometric methodology was successfully benchmarked against standardised Folin-Ciocalteau spectrophotometric method with a good Pearson correlation (r = 0.821, p < 0.01). Hierarchical Cluster Analysis (HCA) was also applied on electrochemical results and a good capability to group tested beers based on their tyrosol concentration was demonstrated. Graphical abstract fx1 Highlights • Screen-printed amperometric biosensor based on AuNPs and tyrosinase. • Beer brewing classification based on tyrosol content. • Potential for field process analysis in the brewing industry. [ABSTRACT FROM AUTHOR]

The development of new techniques for rapid and continuous monitoring of urea in biomedical and clinical analysis is very important. Thus, conductive polymer-supported carbon nanotube as an effective electrochemical biosensing platform for direct detection of urea in blood samples was designed. For the assay optimization, several conductive polymers were synthesized and tested as electrode modifiers; among the tested polymers, poly-o-toluidine (PoT) showed the highest electrochemical signals. However, after the enzyme immobilization, direct bioelectrochemical signals were not obtained when the PoT was used alone. Due to the lower electrocatalytic feature of PoT, integration of carbon nanotube, to form a composite with the PoT, was exploited to enable the direct electron transfer. Successfully, using the hybrid, the catalytic activity of the immobilized urease enzyme was retained. Consequently, a sensitive and specific chronoamperometric signals were achieved after the bioassay optimization. Eventually, a standard calibration curve for urea determination was obtained. A linear range was found from 0.1 to 11 mM with the limit of detection of 0.03 mM. Successfully, several blood samples were analyzed and urea level was correlated with the reference analytical method.Scheme 1: Schematic representation of screen-printed electrode-modified urease biosensors. Three-layer functionalized surface (multi-walled carbon nanotube/poly(o-toluidine) nanocomposite, with glutaraldehyde (cross-linker and urease-immobilized enzyme), was designed for urea biosensors. [ABSTRACT FROM AUTHOR]

The working area of a screen-printed electrode, SPE, was modified with the enzyme tyrosinase (Tyr) using different immobilization methods, namely entrapment with water-soluble polyvinyl alcohol (PVA), cross-linking using glutaraldehyde (GA), and cross-linking using GA and human serum albumin (HSA); the resulting electrodes were termed SPE/Tyr/PVA, SPE/Tyr/GA and SPE/Tyr/HSA/GA, respectively. These biosensors were characterized by means of amperometry and EIS techniques. From amperometric evaluations, the apparent Michaelis-Menten constant, Km′, of each biosensor was evaluated while the respective charge transfer resistance, Rct, was assessed from impedance measurements. It was found that the SPE/Tyr/GA had the smallest Km′ (57 ± 7) µM and Rct values. This electrode also displayed both the lowest detection and quantification limits for catechol quantification. Using the SPE/Tyr/GA, the Trolox Equivalent Antioxidant Capacity (TEAC) was determined from infusions prepared with “mirto” (Salvia microphylla), “hHierba dulce” (Lippia dulcis) and “salve real” (Lippia alba), medicinal plants commonly used in Mexico.

Adequate selection of the electrode surface and the strategies for its modification to enable subsequent immobilization of biomolecules and/or nanomaterials integration play a major role in the performance of electrochemical affinity biosensors. Because of the simplicity, rapidity and versatility, electrografting using diazonium salt reduction is among the most currently used functionalization methods to provide the attachment of an organic layer to a conductive substrate. This particular chemistry has demonstrated to be a powerful tool to covalently immobilize in a stable and reproducible way a wide range of biomolecules or nanomaterials onto different electrode surfaces. Considering the great progress and interesting features arisen in the last years, this paper outlines the potential of diazonium chemistry to prepare single or multianalyte electrochemical affinity biosensors on screen-printed electrodes (SPEs) and points out the existing challenges and future directions in this field. [ABSTRACT FROM AUTHOR]

Clusterin (CLU) has been associated with the clinical progression of Alzheimer's disease (AD) and described as a potential AD biomarker in blood plasma. Due to the enormous attention given to cerebrospinal fluid (CSF) biomarkers for the past couple of decades, recently found blood-based AD biomarkers like CLU have not yet been reported for biosensors. Herein, we report the electrochemical detection of CLU for the first time using a screen-printed carbon electrode (SPCE) modified with 1-pyrenebutyric acid N-hydroxysuccinimide ester (Pyr-NHS) and decorated with specific anti-CLU antibody fragments. This bifunctional linker molecule contains succinylimide ester to bind protein at one end while its pyrene moiety attaches to the carbon surface by means of π-π stacking. Cyclic voltammetric and square wave voltammetric studies showed the limit of detection down to 1 pg/mL and a linear concentration range of 1-100 pg/mL with good sensitivity. Detection of CLU in spiked human plasma was demonstrated with satisfactory recovery percentages to that of the calibration data. The proposed method facilitates the cost-effective and viable production of label-free point-of-care devices for the clinical diagnosis of AD. [ABSTRACT FROM AUTHOR]

This review (with 35 references) summarizes the various strategies used in biosensors for galactose, and their analytical performance. A brief comparison of the enzyme immobilization methods employed and the analytical performance characteristics of a range of galactose biosensors are first summarized in tabular form and then described in detail. Selected examples have been included to demonstrate the various applications of these biosensors to real samples. Following an introduction into the field that covers the significance of sensing galactose in various fields, the review covers biosensors based on the use of galactose oxidase, with a discussion of methods for their immobilization (via cross-linking, adsorption, covalent bonding and entrapment). This is followed by a short section on biosensors based on the use of galactose dehydrogenase. The conclusion section summarizes the state of the art and addresses current challenges. [ABSTRACT FROM AUTHOR]

In this paper for the first time we successfully detect bacterial flagellar filaments from Proteus mirabilis using molecularly imprinted artificial receptors. These receptors acted as a sensing layer of the biosensors, assembled by imprinting flagellar proteins onto a polymeric backbone of electropolymerized phenol. In short, flagellar filaments were absorbed onto a carbon support, phenol was electropolymerized around it through the carbon conductive matrix to create the protein molecular molds, and finally the flagellar proteins were removed by enzymatic and electrochemical action. Each removed flagellar protein gave rise to an imprinted site with eventual rebinding ability. Electrical impedance spectroscopy (EIS) and square wave voltammetry (SWV) were employed to measure the interaction of flagellar filaments with the sensing layer assembled on commercial screen-printed electrodes, providing low detection limits, high precision and selectivity toward the targeted protein. The detection limit was 0.7 ng/mL by EIS and 0.9 ng/mL by SWV. The artificial receptors were further assembled on home-made paper-printed electrodes, with the three-electrode system printed on a paper substrate, offering the possibility of detecting flagellar filaments at as low as 0.6 ng/mL with a disposable and cost-effective portable device. To the best of our knowledge this is the first sensing device where molecularly imprinted artificial receptors are tailored on home-made electrode based on paper substrates with three electrodes assembled together, which is a suitable approach for the fabrication of easy and cost-effective tailored electrodes. [ABSTRACT FROM AUTHOR]

In this work, we proposed a biosensor for trypsin proteolytic activity assay using immobilization of model peptides on screen-printed electrodes (SPE) modified with gold nanoparticles (AuNPs) prepared by electrosynthetic method. Sensing of proteolytic activity was based on electrochemical oxidation of tyrosine residues of peptides. We designed peptides containing N-terminal cysteine residue for immobilization on an SPE, modified with gold nanoparticles, trypsin-specific cleavage site and tyrosine residue as a redox label. The peptides were immobilized on SPE by formation of chemical bonds between mercapto groups of the N-terminal cysteine residues and AuNPs. After the incubation with trypsin, time-dependent cleavage of the immobilized peptides was observed by decline in tyrosine electrochemical oxidation signal. The kinetic parameters of trypsin, such as the catalytic constant (k cat), the Michaelis constant (K M) and the catalytic efficiency (k cat / K M), toward the CGGGRYR peptide were determined as 0.33 ± 0.01 min−1, 198 ± 24 nM and 0.0016 min−1 nM−1, respectively. Using the developed biosensor, we demonstrated the possibility of analysis of trypsin specificity toward the peptides with amino acid residues disrupting proteolysis. Further, we designed the peptides with proline or glutamic acid residues after the cleavage site (CGGRPYR and CGGREYR), and trypsin had reduced activity toward both of them according to the existing knowledge of the enzyme specificity. The developed biosensor system allows one to perform a comparative analysis of the protease steady-state kinetic parameters and specificity toward model peptides with different amino acid sequences. [Display omitted] • An electrochemical biosensor for trypsin activity assay was developed. • Model peptides were immobilized on the electrodes modified with gold nanoparticles. • The assay is based on cleavage of the immobilized tyrosine-containing peptides with unmodified natural amino acid residues. • The activity was monitored by registration of electrooxidation of tyrosine residues. • The biosensor can be used for determination of protease specificity. [ABSTRACT FROM AUTHOR]

Herein, we report the first paper-based microfluidic device encompassing the filter paper to manage the flow and to load the reagents for a reagent-free competitive magnetic-bead-based immunosensor for cortisol analysis in sweat. The paper-based microfluidic pattern was made using wax printing and laser-cutter techniques, for the delivery of capillary-driven microfluidics. The presence of magnetic beads functionalized with monoclonal antibodies for the recognition of the cortisol in the reaction zone allows for the specific measurement of the target analyte. The competitive reaction between the target cortisol and the labeled cortisol with acetylcholinesterase enzyme gives a response inversely proportional to the target cortisol in the range of 10 and 140 ng/mL, by simply folding the pad loaded with the enzymatic substrate. The paper-based microfluidic device was successively combined with a Near-Field Communication wireless module to develop a flexible integrated wearable analytical tool for cortisol detection in sweat. The paper-based integrated device was successfully applied to determine the level of cortisol in sweat in one volunteer during cycling activities at two different times, demonstrating the reliability of this sustainable paper-based device. • Wearable electrochemical paper-based immunosensor for cortisol detection. • Sweat analysis by exploiting filter paper for capillary-driven flow and reagents loading. • Origami configuration for immunoassay without washing step and reagent addition. • Wireless and battery-free wearable device. [ABSTRACT FROM AUTHOR]

Herein, we report a portable electrochemical biosensor based on butyrylcholinesterase (BChE) immobilized on carbon black (CB)-modified screen-printed electrodes (SPEs) for the detection of organophosphorous pesticides in olive oil. The BChE/CB-SPE biosensor was developed to detect paraoxon in standard solutions as well as in olive oil samples previously treated with the QuEChERS method to extract pesticides from the whole fatty matrix. The biosensor shows a linear concentration range of between 20 and 100 ppb for paraoxon both in standard solutions (phosphate buffer 0.05 M) and in olive oil extracts, with a detection limit of 6 ppb in olive oil extract, corresponding to 10% of inhibition. The accuracy of this biosensor in olive oil samples was assessed with olive oil spiked with paraoxon, obtaining satisfactory recovery values. [ABSTRACT FROM AUTHOR]

The development of screen printed sensors and biosensors is a significant area of contemporary electroanalysis because of their wide potential possibilities of practical applications. A particular progress in their development in the recent decade was brought by the application of carbon and metallic nanomaterials, as well as oxide and composite ones. This concerns both chemical sensors, and also enzymatic biosensors, immunosensors, aptasensors and genosensors. This review article, based on 140 selected original works, presents the progress gained in recent years in design of sensors used with voltammetric, potentiometric and impedimetric detections, with brief mentioning also resistance-based gas sensors [ABSTRACT FROM AUTHOR]

A novel amperometric biosensor for the determination of Al(III) based on the inhibition of the enzyme superoxide dismutase has been developed. The oxidation signal of epinephrine substrate was affected by the presence of Al(III) ions leading to a decrease in its amperometric current. The immobilization of the enzyme was performed with glutaraldehyde on screen-printed carbon electrodes modifiedwith tetrathiofulvalene (TTF) and different types of nanoparticles. Nanoparticles of gold, platinum, rhodium and palladium were deposited on screen printed carbon electrodes by means of two electrochemical procedures. Nanoparticles were characterized trough scanning electronic microscopy, X-rays fluorescence, and atomic force microscopy. Palladium nanoparticles showed lower atomic force microscopy parameters and higher slope of aluminum calibration curves and were selected to perform sensor validation. The developed biosensor has a detection limit of 2.0 ± 0.2 μM for Al(III), with a reproducibility of 7.9% (n = 5). Recovery of standard reference material spiked to buffer solution was 103.8% with a relative standard deviation of 4.8% (n = 5). Recovery of tap water spiked with the standard reference material was 100.5 with a relative standard deviation of 3.4% (n = 3). The study of interfering ions has also been carried out. [ABSTRACT FROM AUTHOR]

Ultrasensitive biosensing is currently a growing demand that has led to the development of numerous strategies for signal amplification. In this context, the unique properties of magnetic particles; both of nano- and micro-size dimensions; have proved to be promising materials to be coupled with disposable electrodes for the design of cost-effective electrochemical affinity biosensing platforms. This review addresses, through discussion of selected examples, the way that nano- and micro-magnetic particles (MNPs and MMPs; respectively) have contributed significantly to the development of electrochemical affinity biosensors, including immuno-, DNA, aptamer and other affinity modes. Different aspects such as type of magnetic particles, assay formats, detection techniques, sensitivity, applicability and other relevant characteristics are discussed. Research opportunities and future development trends in this field are also considered. [ABSTRACT FROM AUTHOR]

Polybrominated diphenyl ethers (PBDEs) are persistent environmental substances that were commonly used as fire retardants in a wide number of commercial products. Their low reactivity, high hydrophobicity and bioaccumulative properties cause their ubiquity in the air, water, food and lead to extensive exposure of world population to these compounds. The severe health problems caused by PBDEs lead them to be banned from the market. In March 2014 the European Commission issued a recommendation in which member states are requested to monitor brominated flame retardants in food, in order to evaluate human and wildlife exposure. Here, we described the development of an electrochemical magnetic particle enzyme-linked immunoassay to analyze PBDEs in food samples. The immunological reaction is based on a competitive scheme, using an alkaline phosphatase labeled congener as tracer. The anti-PBDE antibody modified magnetic particles are captured on the surface of carbon disposable array of sensors. The reaction extent is finally electrochemically measured by differential pulse voltammetry, upon the addition of substrate. Under the optimized conditions, a limit of detection of 0.18 ng/mL with a limit of quantification of 0.30 ng/mL and a quantification range of 0.30-6.9 ng/mL, (RSD%=12) is obtained. Results of food samples obtained from the newly developed electrochemical immunoassay are also reported. [ABSTRACT FROM AUTHOR]

An ultra-sensitive screen-printed biosensor was successfully developed for phosmet detection in olive oil, based on a genetically-engineered acetylcholinesterase (AChE) immobilized in a azide-unit water-pendant polyvinyl alcohol (PVA-AWP)/Fe–Ni alloy nanocomposite. Fe–Ni not only allowed amplifying the response current but also lowering the applied potential from 80 mV to 30 mV vs Ag/AgCl. The biosensor showed a very good analytical performance for phosmet detection, with a detection limit of 0.1 nM. This detection limit is lower than the allowable concentrations set by international regulations. In addition to the good reproducibility, operational and storage stability, the developed biosensor was successfully used for the determination of phosmet in olive oil samples without any laborious pre-treatment. The phosmet recovery rate was about 96% after a simple liquid–liquid extraction. [ABSTRACT FROM AUTHOR]

This review addresses recent advances in the development of screen-printed electrode based biosensors modified with different nanomaterials such as carbon nanotubes, graphene, metallic nanoparticles as gold, silver and magnetic nanoparticles, and mediator nanoparticles (Prussian Blue, Cobalt Phthalocyanine, etc.), coupled with biological recognition elements such as enzymes, antibodies, DNA and aptamers to obtain probes with improved analytical features. Examples of clinical applications are illustrated, together with examples of paper-based electrochemical devices, of multiple detections using arrays of screen printed electrodes, and of the most recent developments in the field of wearable biosensors. Also the use of smartphones as final detectors is briefly depicted. [ABSTRACT FROM AUTHOR]

Professor Marco Mascini was an internationally acknowledged scientist, teacher of generations of chemists. He passed away on May 12 2015 at the age of 75. During his scientific carrier he made outstanding and innovative contributions to analytical chemistry, especially in the interdisciplinary field of sensors and biosensors. This paper presents an overview of Prof. Mascini's scientific interests and accomplishments in his career that testify the pivotal role of this scientist in the modern analytical chemistry. [ABSTRACT FROM AUTHOR]

In the past few years a large effort is being made aiming at the development of fast and reliable tests for cancer biomarkers. Protein imprinted sensors can be a fast and reliable strategy to develop tailor made sensors for a large number of relevant molecules. This work aims to produce, optimize and use in biological samples a biosensor for microseminoprotein-beta (MSMB). Caffeic acid (CAF) electropolimerization was performed in the presence of microseminoprotein-beta (MSMB) creating target protein specific cavities on the surface of a screen-printed carbon. Dopamine was introduced as charged monomer labelling the binding site and was allowed to self-organize around the protein. The subsequent electropolimerization was made by applying a constant potential of +2.0 V, for 30 s, on a carbon screen-printed electrode, immersed in a solution of protein and CAF prepared in phosphate buffer. The sensor with charged monomers showed a more sensitive response, with an average slope of −7.59 μA/decade, linear concentration range of 0.5–100 ng/mL and a detection limit of 0.12 ng/mL. The corresponding non-imprinted sensor displayed an inconsistent response over the range of the calibration curve. The biosensor was successfully applied to the analysis of MSMB in serum and urine samples. [ABSTRACT FROM AUTHOR]

Here we describe the construction and optimization of a cholesterol biosensor based on screen-printed electrodes (SPEs) modified with inkjet-printed Prussian blue nanoparticles (PBNPs). The deposition of PBNPs using inkjet printing led to the highly facile fabrication of sensors with excellent sensitivity and reproducibility for the measurement of H 2 O 2 . Further integration of the sensor with a microfabricated low volume (4 μL) sample cell allowed the measurement of cholesterol in serum with the addition of cholesterol oxidase. The biosensor exhibited a sensitivity to cholesterol of 2.1 μA/mM cm 2 ( r 2 = 0.97, n = 5) and was linear in the range of 0–15 mM. [ABSTRACT FROM AUTHOR]

Combining the advantages of the biosensor field with the problem of detecting Human Cytomegalovirus (HCMV) in human samples, an inexpensive, simple and disposable electrochemical immunosensor for glycoprotein B detection in urine is proposed. Glycoprotein B has been chosen once is the dominant antigen of HCMV. The approach is based on a sandwich-type immunoassay, with the HCMV glycoprotein B sandwiched between the Anti-HCMV antibody adsorbed onto the working electrode, and the Anti-HCMV labeled with gold nanoparticles. Glycoprotein B detection was carried out through electrochemical stripping analysis of silver nanoparticles quantitatively deposited on the immunosensor through catalysis by the nanogold labels. The influence of different steps in the immunosensor construction, namely the silver deposition time, silver enhancer concentration, antibody concentration, BSA concentration and glycoprotein B incubation time, were examined and optimized. The method showed a linear dependence between glycoprotein B concentration and the corresponding anodic stripping peak current, resulting in detection limits of 3.3±1.7 ng/mL for samples prepared in buffer and 3.2±0.2 ng/mL for urine samples, suggesting that the biological matrix does not interfere with the immunosensor detection capability. Given its mode of preparation, by physical adsorption of the capture antibody in the working electrode, the immunosensor also exhibited an acceptable reproducibility, with a residual standard deviation of 13.5% for samples prepared in buffer and 11.2% for urine samples, thereby presenting a promising development potential for clinical applications. [ABSTRACT FROM AUTHOR]

• Monitoring of astringent compounds in food beverages by means of an electrochemical biosensor. • Biosensor assembled on gold screen-printed electrodes for analysis in-situ. • Interaction between polyphenols and salivary proteins monitored by electrochemistry. • Sensitivity of the electrochemical responses depends on the salivary protein. • Successful application to wine sample analysis, in line with the sensory panel. An innovative approach for monitoring astringent polyphenols in beverages (wines) is described, consisting of an electrochemical biosensor constructed by adsorbing salivary α-amylase or proline-rich protein (PRP) onto amined gold screen-printed electrodes. Interaction with polyphenols was tested using pentagalloyl glucose (PGG) as a standard, an important representative element for astringency. The analytical properties of the resulting biosensors were evaluated by electrochemical impedance spectroscopy at different pHs. The PRP-biosensor was able to bind to PGG with higher sensitivity, displaying lower limit of the linear range of 0.6 µM. Wine samples were tested to prove the concept and the concentrations obtained ranged from 0.17 to 4.7 µM, as expressed in PGG units. The effects of side-compounds on PRP and on α-amylase binding to PGG were tested (gallic acid, catechin, ethanol, glucose, fructose and glycerol) and considered negligible. Overall, concentrations > 1.0 µM in PGG units are signaling electrochemical impedance, providing a quantitative monitoring of astringent compounds. [ABSTRACT FROM AUTHOR]

Putrescine oxidase (PUO) from Micrococcus rubens has been characterized, as a biocatalytic component for the development of electrochemical biosensors for the determination of putrescine. PUO was immobilized by cross-linking on screen-printed carbon electrodes (SPCEs) containing tetrathiafulvalene (TTF) as mediator (TTF/SPCEs). Isothermal titration calorimetry was used for investigation of kinetic properties of immobilized PUO. These PUO/TTF/SPCEs showed a linear response range from 10 a 74 µM, a capability of detection of 10.1±0.6 µM ( n=4, α= β=0.05) and a precision of 6.7 %. Octopus and zucchini samples have been analysed using HPLC as a reference technique. [ABSTRACT FROM AUTHOR]

A new polymer composition was developed based on hybrid sol-gel material for the immobilization of enzymes on the surface of screen-printed carbon electrodes modified with Prussian blue. The existing and developed methods for the glucose oxidase (GO) immobilization are compared. Highly stable bioelectrodes were shown to be obtained by enzyme immobilization on a hybrid composition consisting of solgel/poly vinyl alcohol (PVA) (up to 60 sequential chains) and agar gel (up to 45 sequential chains). The range of glucose concentrations detected during enzyme immobilization in a hybrid sol-gel/PVA composition or agar gel without dilution of a sample was 1.0-5.9 μM and 3.6-6.3 μM, respectively. An analysis of wine products was conducted. The results obtained using the proposed biosensors were shown to differ insignificantly from those obtained by high-performance liquid chromatography (the correlation coefficient was 0.9998). [ABSTRACT FROM AUTHOR]

The working area of a screen-printed electrode, SPE, was modified with the enzyme tyrosinase (Tyr) using different immobilization methods, namely entrapment with water-soluble polyvinyl alcohol (PVA), cross-linking using glutaraldehyde (GA), and cross-linking using GA and human serum albumin (HSA); the resulting electrodes were termed SPE/Tyr/PVA, SPE/Tyr/GA and SPE/Tyr/HSA/GA, respectively. These biosensors were characterized by means of amperometry and EIS techniques. From amperometric evaluations, the apparent Michaelis-Menten constant, Km', of each biosensor was evaluated while the respective charge transfer resistance, Rct, was assessed from impedance measurements. It was found that the SPE/Tyr/GA had the smallest Km' (57 ± 7) µM and Rct values. This electrode also displayed both the lowest detection and quantification limits for catechol quantification. Using the SPE/Tyr/GA, the Trolox Equivalent Antioxidant Capacity (TEAC) was determined from infusions prepared with "mirto" (Salvia microphylla), "hHierba dulce" (Lippia dulcis) and "salve real" (Lippia alba), medicinal plants commonly used in Mexico. [ABSTRACT FROM AUTHOR]

Abstract: A label-free electrochemical impedance spectroscopy biosensor for selective detection and discrimination of the cancer-associated sialyl-Tn (STn) antigen was developed by using Sambucus nigra agglutinin type I (SNA-I) as the recognition element. The SNA-I biosensor was constructed by immobilizing the lectin on screen-printed gold electrodes. The formation of a complex between SNA-I and STn-containing glycoproteins (transferrin and bovine submaxillary mucin) was monitored by measuring the impedance increase of the biosensor. The increase in electron transfer resistance was linearly proportional to the concentration of the glycoproteins up to 70ng of transferrin and 40ng of bovine submaxillary mucin, with a limit of detection of 20ng for transferrin. Albumin, the most abundant serum protein, did not interfere in the detection of the STn-glycoproteins up to a concentration of 0.2mgml−1. The developed lectin-based biosensor was used to evaluate the STn-expression in serum samples and discriminate samples from healthy individuals and patients with different types of malignant tumors, mostly carcinomas, where the increased expression of STn aberrant glycans is well established. This work demonstrates the feasibility of employing SNA-I to selectively recognize the STn epitope in glycoproteins and the use of the constructed biosensor was effective in the analysis of serum samples with the ability to discriminate in a fast way between cancer and healthy status. The proposed biosensor could be used for high-throughput, label-free profiling of the cancer-associated STn glycan expression in serum for diagnosis and therapy monitoring. [Copyright &y& Elsevier]

Highlights: [•] Chronoamperometric determination of Cr(III) on tyrosinase based biosensors using SPCEs. [•] Chronoamperometric determination of Cr(VI) on GOx based biosensors using SPCEs. [•] High degree of sensitivity and selectivity in the analysis of both chromium species. [•] Bipotentiostatic chronoamperometric determination of both chromium species in the same sample. [Copyright &y& Elsevier]

This work describes a novel concept of biosensor for quantifying enzymes, where the substrate is immobilized directly over the working area of a screen printed electrode (Au-SPE). This concept is applied here to creatine kinase isoenzyme (CK-MB), a cardiac biomarker in ischemic conditions. It acts as a phospho-transferase on creatine (Crea), requiring the presence of phosphate. So, the phosphorylated form of creatine (Pcrea) was immobilized on the Au/SPE previously aminated with cysteamine (Cys) by self-assembling monolayer technique. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) studies were used to follow the chemical modifications in the Au-SPE. Since Pcrea is an electroactive species at low potential, its consumption over the platform by the enzyme changed the electrical response of the biosensor. So, CK-MB determination has been achieved in mediator free-conditions due the redox proprieties of the Pcrea. The analytical features of the resulting biosensor were studied by square wave voltammetry (SWV). The limit of detection was 0.11µg/mL and the slope was −0.029(±0.0035)µA×mL/µg. The interference effect of troponin T (TnT), bovine serum albumin (BSA) and myoglobin (Myo) in the performance of the sensor was tested and good selectivity was observed. The biosensor was successfully applied to biological fluids, showing good stability at room temperature and excellent sensitivity and selectivity. This new concept of biosensor is especially useful for point of care (POC) applications, due to the low cost and small size of the final device. [ABSTRACT FROM AUTHOR]

Highlights: [•] First disposable amperometric magnetobiosensor for screening of multiple antibiotics. [•] Discrimination between free and milk samples containing antibiotics at their MRLs. [•] Simple and short sample pretreatment and fast detection within 5min. [•] Useful and affordable alternative to classical assays for antibiotic screening in milk. [ABSTRACT FROM AUTHOR]

The selective and simultaneous amperometric determination of putrescine (Put) and cadaverine (Cad) has been carried out using a novel design of screen-printed carbon electrode (SPCE) with two working electrodes connected in array mode. A mixture of 3% of tetrathiafulvalene (TTF), as mediator, and carbon ink was used for the construction of the screen-printed working electrode. The employment of different amounts of monoamine oxidase (MAO) enzyme on these modified TTF/SPCEs and the use of gold nanoparticles (AuNPs) allowed performing the simultaneous determination of both analytes. The amperometric detection has been performed by measuring the oxidation current of the mediator at a potential of+250mV vs. screen-printed Ag/AgCl reference electrode. A linear response in the Cad concentration range from 19.6 till 107.1µM and from 9.9 till 74.1μM for Put was obtained at the MAO/AuNPs/TTF/SPCE biosensor. This device showed a capability of detection of 9.9 and 19.9±0.9µM (n=4 α=β=0.05) and a precision of 4.9% and 10.3% in terms of relative standard deviation for Put and Cad, respectively. The developed biosensor was successfully applied to the simultaneous determination of Put and Cad in octopus samples. [ABSTRACT FROM AUTHOR]