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Aggregation of α-synuclein has been recognized as a critical event in the pathogenesis of Parkinson's disease whose prevalence is increasing with great socio-economic challenges for future generations. Here, we developed a sensitive and specific electrochemical immunosensor for the detection and quantification of this biomarker, based on the voltammetric study of a redox indicator signal, which decreases upon the analyte recognition by the antibody due to the electronic resistance increase. The proposed immunosensor is based on a screen-printed carbon electrode modified in a layer-by-layer approach, which through extensive characterization led to the successful nanostructuration of the transducer, through the drop-cast of 3.0 μL of a 0.1 mg mL−1 single-walled carbon nanotubes suspension followed by electrodeposition of gold nanoparticles in a 3 mM HAuCl4 solution under a −0.2 V potential for 150 s. Monoclonal antibodies were immobilized on the gold nanoparticles surface through chemical modification at an optimal concentration of 200 μg mL−1. Using the proposed immunosensor, α-synuclein was detected in the range of 0.01–10 ng mL−1 with a 4.1 and 12.6 pg mL−1 limits of detection and quantification, respectively. Recovery values of 96.7, 106.2 and 102.9% were attained for the tested concentrations spiked in fetal bovine serum while also presenting excellent specificity and stability throughout one month. The nanostructured immunosensor provided a great interface for electronic transduction and biological recognition events, which enabled fast, sensitive and specific detection of α-synuclein while being based on a simple and inexpensive technology requiring small sample volumes, crucial characteristics for application in point-of-care testing. [Display omitted] • A simple and effective label-free procedure was developed for α-synuclein detection. • The immunosensor is based on a nanostructured screen-printed carbon electrode. • Nanostructuration of the immunosensor amplified its electrochemical behavior. • Detection and quantification limits of 4.1 and 12.6 pg mL−1 were attained. • The immunosensor enables highly sensitive and specific detection of α-synuclein. [ABSTRACT FROM AUTHOR]

A novel electrochemical sensor was developed for the detection of salivary cortisol levels. The sensor employs a combination of a molecularly imprinted polymer (MIP) and gold nanoparticles (AuNPs) that are electrodeposited onto a screen-printed electrode (SPE). The study utilised density functional theory and molecular docking techniques to determine the geometry of molecular orbitals, electrostatic potential energies, and binding energy of cortisol and the polymers. The thin film of cortisol-imprinted polymer on the SPE was created by electro-polymerizing pyrrole and thiophene-3-carboxylic acid on the electrode surface along with cortisol as the template molecule. The MIP film was characterised using scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), and electrochemical techniques. The sensor exhibited a linear response in the concentration range of 0.05 nmol L−1 to 2.5 μmol L−1, with a limit of detection of 0.01 nmol L−1, as determined by differential pulse voltammetry. This method offers a simple yet efficient and sensitive approach to detecting cortisol levels in human saliva samples. [Display omitted] • A new electrochemical sensor is presented that can selectively detect salivary cortisol levels using a molecularly imprinted polymer (MIP). • Gold nanoparticles (AuNPs) were electrochemically deposited on a screen-printed electrode (SPE) as a substrate for the cortisol-imprinted polymer thin film. • The cortisol-imprinted polymer thin film was synthesized on the SPE through the electro-polymerization of pyrrole and thiophene-3-carboxylic acid. • The study used density functional theory and molecular docking techniques to compute the geometry of molecular orbitals, electrostatic potential energies, and binding energy for the interacting molecules. • The differential pulse voltammetry method showed a linear response in the concentration range of 0.05 nmol L−1 to 2.5 μmol L−1 with a limit of detection of 0.016 nmol L−1. • This simple approach is an efficient and sensitive method for detecting cortisol levels in human saliva samples. [ABSTRACT FROM AUTHOR]

The availability of more efficient analytical methods that answer the world's demands is a challenge and their development continues to be a difficult task. In this work the construction of an electrochemical cell, based on low-cost and accessible materials, that can be easily constructed and used for electroanalytical purposes, is described. Pencil leads were used as electrodes and a transparency sheet as the base. This cell was used as transducer for developing an amperometric biosensor for the quantification of histamine, which is the only biogenic amine regulated by law. The analysis was based on the use of diamine oxidase as biorecognition element, hexacyanoferrate(III) as electron-transfer mediator, and chronoamperometry, at +0.5 V during 100 s, to record the analytical signal. A linear relationship between histamine concentration and the analytical signal was established between 5.0 and 35 mg L−1 and a low limit of detection (1.0 mg L−1) was achieved. The analysis of different fish species (sardine and tuna) was performed, obtaining recovery values between 102% and 110%. The stability of the sensor is noteworthy: it maintained 95% of the initial analytical signal after 15 days. [Display omitted] • A simple electrochemical cell consisting of three graphite pencil leads was created. • A cheap histamine biosensor using 0.5-mm pencil leads as electrodes was developed. • The sensor showed satisfactory linear range (5–35 mg L−1) and reproducibility (<4%). • The sensor showed suitable stability (95% signal after 15 days) and selectivity. • The sensor provided good recoveries when applied to spiked fish samples. [ABSTRACT FROM AUTHOR]

Cardiovascular diseases are among the major causes of mortality and morbidity. Warfarin is often prescribed for these disorders, an anticoagulant with inter and intra-dosage variability dose required to achieve the target international normalized ratio. Warfarin presents a narrow therapeutic index, and due to its variability, it can often be associated with the risk of hemorrhage, or in other patients, thromboembolism. Single-nucleotide polymorphisms are included in the causes that contribute to this variability. The Cytochrome P450 (CYP) 2C9*3 genetic polymorphism modifies its enzymatic activity, and hence warfarin's plasmatic concentration. Thus, the need for a selective, rapid, low-cost, and real-time detection device is crucial before prescribing warfarin. In this work, a disposable electrochemical DNA-based biosensor capable of detecting CYP2C9*3 polymorphism was developed. By analyzing genomic databases, two specific 78 base pairs DNA probes; one with the wild-type adenine (Target-A) and another with the cytosine (Target-C) single-nucleotide genetic variation were designed. The biosensor implied the immobilization on screen-printed gold electrodes of a self-assembled monolayer composed by mercaptohexanol and a linear CYP2C9*3 DNA-capture probe. To improve the selectivity and avoid secondary structures a sandwich format of the CYP2C9*3 allele was designed using complementary fluorescein isothiocyanate-labeled signaling DNA probe and enzymatic amplification of the electrochemical signal. Chronoamperometric measurements were performed at a range of 0.015–1.00 nM for both DNA targets achieving limit of detection of 42 p.m. The developed DNA-based biosensor was able to discriminate between the two synthetic target DNA targets, as well as the targeted denatured genomic DNA, extracted from volunteers genotyped as non-variant homozygous (A/A) and heterozygous (A/C) of the CYP2C9*3 polymorphism. [Display omitted] • A disposable genosensor able of distinguishing CYP2C9*3 polymorphisms was developed. • The developed sensor was able to discriminate between the two SNP probes. • Genomic DNA samples from fully genotyped volunteers were analyzed. • The developed analytical device is a promising alternative to genotyping methodologies. [ABSTRACT FROM AUTHOR]

Specific monitoring of cystatin C (CysC) levels in biological fluids is critical for diagnosis, treatment and mechanistic understanding of a spectrum of diseases, particularly chronic kidney disease (CKD). Despite evidences that CysC correlates with the high risk and/or progression of CKD, its use in clinical practice is still scarce. In this context, we report the development of a simple and sensitive immunosensor for the detection of CysC. The biosensor combines the technology of cost-effective screen-printed electrodes with the high specificity of a sandwich immunoassay. Optimized conditions showed that the sensor operates in a linear range between 10 and 100 ng mL−1, with a detection limit and a sensitivity of 6.0 ng mL−1 and 6.4 ± 0.3 μA ng mL−1 cm−2, respectively. Moreover, the sensor provided precise results (RSD ≤ 6.2%) and the quantification of CysC in CKD serum samples revealed to be in agreement with the values obtained by a particle-enhanced nephelometric immunoassay. In this light, the proposed immunosensor qualifies for clinical application, constituting a step forward in the development of fast, sensitive and cost-effective diagnostic tools that can improve the current medical care settings of CKD patients. Image 1 • An immunosensor for CysC, a chronic kidney disease biomarker, was developed. • Screen-printed carbon electrodes nanostructured with gold nanoparticles were used. • The sensor showed a linear concentration range with clinical utility. • The sensor was applied to CysC quantification in serum samples from CKD patients. [ABSTRACT FROM AUTHOR]

Abstract Recent studies have shown that besides the well-known T 3 (triiodothyronine) and T 4 (thyroxine) there might be other important thyroid hormones, in particular T 0 AM (thyronamine) and T 1 AM (3-iodothyronamine). The absence of a large number of studies showing their precise importance might be explained by the limited number of analytical methodologies available. This work aims to show an electroanalytical alternative making use of electropolymerized molecularly imprinted polymer (MIPs). The MIPs' polymerization is performed on the surface of screen-printed carbon electrodes (SPCEs), using 4–aminobenzoic acid (4-ABA) as the building and functional monomer and the analyte T 0 AM as the template. The step-by-step construction of the SPCE-MIP sensor was studied by cyclic voltammetry (CV) and by electrochemical impedance spectroscopy (EIS). After optimization, by means of square-wave voltammetry, the SPCE-MIP showed suitable selectivity (in comparison with other thyroid hormones and catechol amines), repeatability (intra-day of 3.9%), a linear range up to 10 μmol L−1 (0.23 × 103 μg dL−1) with an r2 of 0.998 and a limit of detection (LOD) and quantification (LOQ) of 0.081 and 0.27 μmol L−1 (1.9 and 6.2 μg dL−1), respectively. Graphical abstract fx1 Highlights • First time a sensor was developed for T 0 AM (thyronamine); • Electroanalytical methodology for T 0 AM; • Molecularly imprinted polymers (MIPs) were developed for T 0 AM; • MIPs' polymerization (4–aminobenzoic acid to a poly(4-aminobenzoic acid) film) occurred in the surface of screen-printed carbon electrodes (SPCEs); • The SPCE-MIP selectively analyzed T 0 AM by square-wave voltammetry (SWV). [ABSTRACT FROM AUTHOR]

Abstract Polymeric ion selective electrodes are highly sensitive to changes in zero current ion flow and this offers a route to signal amplification in label-free potentiometric immunosensors. In this work, a label-free potentiometric immunosensor toward Salmonella typhimurium (ST) assembled in a home-made pipette-tip electrode is described. The signal-output amplification was implemented on a gold nanoparticle polymer inclusion membrane (AuNPs-PIM) which was used as sensing platform and for antibody immobilization. Additionally, a marker ion was used to detect the antibody-antigen binding event at the electrode surface. The immunosensor construction was performed in several steps: i) gold salt ions extraction in PVC membrane; ii) AuNPs formation using Na 2 EDTA as reduction agent; iii) antibody anti- Salmonella conjugation on AuNPs-PIM in pipette-tip electrodes. The potential shift observed in potentiometric measurements was derived simply from the blocking effect in the ionic flux caused by antigen-antibody conjugation, without no extra steps, mimetizing the ion-channel sensors. A detection limit of 6 cells mL−1 was attained. As proof-of-concept, recovery studies were performed in spiked commercial apple juice samples with success. Due to the simplicity of use, the appealing cost of equipment and sensor production and being able to provide a quick analytical response (less than 1 h for a complete assay, including sample preparation for analysis), this scheme represents a good prototype device for the detection of foodborne pathogens like ST or other immune-responsive bacteria. Graphical abstract fx1 Highlights • A label free sensor for Salmonella , based on surface blocking principle, is proposed. • Au nanoparticle polymer inclusion membrane is the antibody support and a sensor platform. • A detection limit of 6 cells mL−1 is attained. [ABSTRACT FROM AUTHOR]

The remarkable features of a carbon fiber paper sensor (CP) were employed for detection of the estrogenic hormone 17α-ethinylestradiol (EE2), considered a contaminant of emerging concern due to its potential ecotoxicity and widespread in the aquatic ecosystems. In this work, an unpreceded CP pre-treatment study was conducted with the (Il)-hexacyanoferrate(III) ion pair, however a bare sensor without pre-treatment revealed higher efficiency on the oxidation of EE2 compared to a chemical and electrochemical pre-treated CP and a gold nanoparticles modified CP, being thus selected for EE2 determinations. The analytical conditions were thoroughly optimized in terms of electrolyte pH (pH 7), differential pulse voltammetry parameters (modulation time 0.003 s, amplitude 0.09 V, interval time 0.1 s and step potential 0.01 V), and analyte preconcentration potential (0.4 V) and time (180 s). The hormone can be determined by the CP in a wide linear range from 0.1 to 1000 nM, achieving a detection limit of 0.14 ± 0.005 nM and an outstanding sensitivity of 1636 ± 232 μA μM−1 cm−2 in the lowest linear zone (0.1–1 nM). The sensor was validated in river water and fish reaching good recoveries (91.2 ± 4.6 to 109.0 ± 7.1%), reproducibility and repeatability. Moreover, the sensor showed high selectivity to EE2 in the presence of several potential interfering compounds and frequently prescribed drugs, though it could not discriminate the similar hormone, 17β-estradiol, being the total concentration obtained in this case. CP-based sensors emerge as efficient electroanalytical tools, suggesting that modification of the surface may not always be beneficial in terms of sensitivity. [Display omitted] • A simple and effective carbon paper sensor was used for 17α-ethinylestradiol sensing. • The best analytical signal was obtained with an untreated and unmodified carbon paper. • The optimized sensor achieved high sensitivity (1636 μA μM−1 cm−2) and LOD of 0.14 nM. • Application in complex samples of fish resulted in recoveries from 102 to 109%. • Carbon paper sensors can be become valuable tools in the electroanalytical field. [ABSTRACT FROM AUTHOR]

In the present work, the development of a genosensor for the event-specific detection of MON810 transgenic maize is proposed. Taking advantage of nanostructuration, a cost-effective three dimensional electrode was fabricated and a ternary monolayer containing a dithiol, a monothiol and the thiolated capture probe was optimized to minimize the unspecific signals. A sandwich format assay was selected as a way of precluding inefficient hybridization associated with stable secondary target structures. A comparison between the analytical performance of the Au nanostructured electrodes and commercially available screen-printed electrodes highlighted the superior performance of the nanostructured ones. Finally, the genosensor was effectively applied to detect the transgenic sequence in real samples, showing its potential for future quantitative analysis. [ABSTRACT FROM AUTHOR]

Human epidermal growth factor receptor 2 (HER2) is a breast cancer biomarker that plays a major role in promoting breast cancer cell proliferation and malignant growth. The extracellular domain (ECD) of HER2 can be shed into the blood stream and its concentration is measurable in the serum fraction of blood. In this work an electrochemical immunosensor for the analysis of HER2 ECD in human serum samples was developed. To achieve this goal a screen-printed carbon electrode, modified with gold nanoparticles, was used as transducer surface. A sandwich immunoassay, using two monoclonal antibodies, was employed and the detection of the antibody-antigen interaction was performed through the analysis of an enzymatic reaction product by linear sweep voltammetry. Using the optimized experimental conditions the calibration curve (ip vs. log[HER2 ECD]) was established between 15 and 100 ng/mL and a limit of detection (LOD) of 4.4 ng/mL was achieved. These results indicate that the developed immunosensor could be a promising tool in breast cancer diagnostics, patient follow-up and monitoring of metastatic breast cancer since it allows quantification in a useful concentration range and has an LOD below the established cut-off value (15 ng/mL). [ABSTRACT FROM AUTHOR]

Abstract: A glutathione-S-transferase (GST) based biosensor was developed to quantify the thiocarbamate herbicide molinate in environmental water. The biosensor construction was based on GST immobilization onto a glassy carbon electrode via aminosilane–glutaraldehyde covalent attachment. The principle supporting the use of this biosensor consists of the GST inhibition process promoted by molinate. Differential pulse voltammetry was used to obtain a calibration curve for molinate concentration, ranging from 0.19 to 7.9mgL−1 and presenting a detection limit of 0.064mgL−1. The developed biosensor is stable, and reusable during 15 days. The GST-based biosensor was successfully applied to quantify molinate in rice paddy field floodwater samples. The results achieved with the developed biosensor were in accordance with those obtained by high performance liquid chromatography. The proposed device is suitable for screening environmental water analysis and, since no sample preparation is required, it can be used in situ and in real-time measurements. [Copyright &y& Elsevier]

This study focused on the development of a sensitive enzymatic biosensor for the determination of pirimicarb pesticide based on the immobilization of laccase on composite carbon paste electrodes. Multi-walled carbon nanotubes (MWCNTs) paste electrode modified by dispersion of laccase (3%, w/w) within the optimum composite matrix (60:40%, w/w, MWCNTs and paraffin binder) showed the best performance, with excellent electron transfer kinetic and catalytic effects related to the redox process of the substrate 4-aminophenol. No metal or anti-interference membrane was added. Based on the inhibition of laccase activity, pirimicarb can be determined in the range 9.90×10−7 to 1.15×10−5 molL−1 using 4-aminophenol as substrate at the optimum pH of 5.0, with acceptable repeatability and reproducibility (relative standard deviations lower than 5%). The limit of detection obtained was 1.8×10−7 molL−1 (0.04mgkg−1 on a fresh weight vegetable basis). The high activity and catalytic properties of the laccase-based biosensor are retained during ca. one month. The optimized electroanalytical protocol coupled to the QuEChERS methodology were applied to tomato and lettuce samples spiked at three levels; recoveries ranging from 91.0±0.1% to 101.0±0.3% were attained. No significant effects in the pirimicarb electroanalysis were observed by the presence of pro-vitamin A, vitamins B1 and C, and glucose in the vegetable extracts. The proposed biosensor-based pesticide residue methodology fulfills all requisites to be used in implementation of food safety programs. [ABSTRACT FROM AUTHOR]

In the present work, the exceptional electrochemical properties of carbon paper were characterized and explored in the development of a sensor for the anti-inflammatory drug, ketoprofen. An initial electrochemical characterization of the carbon paper with a redox indicator revealed much higher voltammetric peaks in comparison with other carbon-based electrodes, namely, screen-printed carbon electrode, boron-doped diamond electrode, glassy carbon electrode, pyrolytic graphite electrode, and pencil graphite electrode, predicting a good sensing performance. Ketoprofen showed a pronounced cathodic peak around −1.1 V vs Ag/AgCl (KCl, 3 M) at pH 5 in Britton-Robinson buffer, presenting a diffusion-controlled process. An optimized differential pulse voltammetry procedure was employed for ketoprofen determination achieving a limit of detection of 0.11 ± 0.01 μM, a linearity up to 6.02 μM, and a high sensitivity of 24 ± 4 or 35 ± 3 μA μM−1 cm−2 (depending on the calibration range: 0.088–1.96 μM or 1.96–6.02 μM, respectively) with acceptable (6–15% relative standard deviation) reproducibility and repeatability considering the challenging conditions of its detection in aqueous solutions. The sensor revealed to be highly selective in the presence of common interferents and other widely consumed anti-inflammatory drugs. Moreover, the developed sensor reached good accuracy in wastewater and fish samples with recoveries varying from 82.3 ± 4.4 to 88.6 ± 4.5%. Image 1 • An unmodified carbon paper was efficiently used as sensor for ketoprofen analysis. • Higher voltammetric peaks were obtained compared to other carbon-based electrodes. • The ketoprofen sensor achieved high sensitivity (35 μA μM cm−2) and low LOD (0.11 μM). • Recoveries of 82.3–88.6% were obtained for complex samples of wastewater and fish. • Carbon paper constitutes a promising sensor for in-situ environmental analysis. [ABSTRACT FROM AUTHOR]

This work addresses the development of a disposable electrochemical genosensor for the detection of the toxic dinoflagellate, Alexandrium minutum. Analyzing public databases, a specific 70 bp DNA probe, targeting A. minutum , was selected and designed. The genosensor methodology implied the immobilization of a A. minutum -specific DNA-capture probe onto screen-printed gold electrodes (SPGE). To improve both the selectivity and to avoid strong secondary structures, that could hinder the hybridization efficiency, a sandwich format of the A. minutum gene was designed using a fluorescein isothiocyanate-labelled signaling DNA probe and enzymatic amplification of the electrochemical signal. Using this electrochemical genosensor, a concentration range from 0.12 to 1.0 nM, a LD of 24.78 pM with a RSD <5.2% was determined. The genosensor was successfully applied to the selective analysis of the targeted A. minutum specific region denatured genomic DNA extracted from toxic dinoflagellates present in the Atlantic Ocean. Image 1 • PCR-free electrochemical biosensing of toxic dinoflagellate nucleic acids. • Electrochemical disposable genosensor for targeting Alexandrium minutum coding sequence. • LOD of 25 pM (0.6 fmol in 25 μL). • Unequivocal detection of A. minutum in Atlantic Ocean water. [ABSTRACT FROM AUTHOR]

This work reports the development of an electrochemical immunosensor for rapid, specific and decentralized detection of the invasion-associated protein p60 secreted by Listeria monocytogenes , a life-threatening foodborne pathogen. A disposable screen-printed electrode was used as transducer surface and monoclonal and polyclonal antibodies that specifically recognize Listeria monocytogenes p60 protein and Listeria spp. p60 proteins, respectively, were used as the sandwich immuno-pair. The reaction was detected with the aid of an additional secondary antibody conjugated with the enzyme reporter (alkaline phosphatase) and using 3-indoxyl phosphate/silver ions as the mixture substrate. The analytical signal was acquired through the voltammetric stripping of the enzymatically deposited silver, which was directly correlated to p60 concentration in the sample. In optimized conditions, a limit of detection and quantification of 1.5 ng mL−1 and 5.1 ng mL−1 were achieved, respectively, in a useful time (<3 h). As proof-of-concept, the proposed immunosensor was successfully applied to spiked milk samples, demonstrating to be a suitable device for further use in real sample detection of Listeria monocytogenes in food products. Image 1 • Listeria monocytogenes is a life-threatening foodborne pathogen. • Invasion-associated protein p60 is a virulence factor secreted by Listeria spp.. • A disposable electrochemical immunosensor for Listeria monocytogenes p60 determination was developed. • The immunosensor achieved a limit of detection of 1.5 ng mL−1 in a useful time (<3 h). • The immunosensing approach accurately detected Listeria monocytogenes p60 in spiked milk samples. [ABSTRACT FROM AUTHOR]

Early detection of cancer increases the possibility for an adequate and successful treatment of the disease. Therefore, in this work, a disposable electrochemical immunosensor for the front-line detection of the ExtraCellular Domain of the Human Epidermal growth factor Receptor 2 (HER2-ECD), a breast cancer biomarker, in a simple and efficient manner is presented. Bare screen-printed carbon electrodes were selected as the transducer onto which a sandwich immunoassay was developed. The affinity process was detected through the use of an electroactive label, core/shell CdSe@ZnS Quantum Dots, by differential pulse anodic stripping voltammetry in a total time assay of 2 h, with an actual hands-on time of less than 30 min. The proposed immunosensor responded linearly to HER2-ECD concentration within a wide range (10–150 ng/mL), showing acceptable precision and a limit of detection (2.1 ng/mL, corresponding to a detected amount (sample volume = 40 μL) of 1.18 fmol) which is about 7 times lower than the established cut-off value (15 ng/mL). The usefulness of the developed methodology was tested through the analysis of spiked human serum samples. The reliability of the presented biosensor for the selective screening of HER2-ECD was confirmed by analysing another breast cancer biomarker (CA15-3) and several human serum proteins. Image 1 • Detection of cancer biomarkers in biological fluids is useful for early diagnosis. • An electrochemical immunosensor was developed for HER2-ECD. • Quantum dots were used as detection label. • The assay was selective towards the target biomarker. • The assay was successfully tested in the analysis of human serum. [ABSTRACT FROM AUTHOR]