Biosensor-based analysis
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The enzyme acetylcholinesterase (AChE) is an important part of cholinergic nervous system, where it stops neurotransmission by hydrolysis of the neurotransmitter acetylcholine. It is sensitive to inhibition by organophosphate and carbamate insecticides, some Alzheimer disease drugs, secondary metabolites such as aflatoxins and nerve agents used in chemical warfare. When immobilized on a sensor (physico-chemical transducer), it can be used for assay of these inhibitors. In the experiments described herein, an AChE- based electrochemical biosensor using screen printed electrode systems was prepared. The biosensor was used for assay of nerve agents such as sarin, soman, tabun and VX. The limits of detection achieved in a measuring protocol lasting ten minutes were 7.41 × 10(-12) mol/L for sarin, 6.31 × 10(-12) mol /L for soman, 6.17 × 10(-11) mol/L for tabun, and 2.19 × 10(-11) mol/L for VX, respectively. The assay was reliable, with minor interferences caused by the organic solvents ethanol, methanol, isopropanol and acetonitrile. Isopropanol was chosen as suitable medium for processing lipophilic samples.
- MeSH
- acetylcholinesterasa chemie MeSH
- analýza selhání vybavení MeSH
- biosenzitivní techniky přístrojové vybavení MeSH
- chemické bojové látky analýza chemie MeSH
- cholinesterasové inhibitory analýza chemie MeSH
- design vybavení MeSH
- konduktometrie přístrojové vybavení MeSH
- neurotoxiny analýza chemie MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
An ultrasensitive impedimetric glycan-based biosensor for reliable and selective detection of inactivated, but intact influenza viruses H3N2 was developed. Such glycan-based approach has a distinct advantage over antibody-based detection of influenza viruses since glycans are natural viral receptors with a possibility to selectively distinguish between potentially pathogenic influenza subtypes by the glycan-based biosensors. Build-up of the biosensor was carefully optimized with atomic force microscopy applied for visualization of the biosensor surface after binding of viruses with the topology of an individual viral particle H3N2 analyzed. The glycan biosensor could detect a glycan binding lectin with a limit of detection (LOD) of 5 aM. The biosensor was finally applied for analysis of influenza viruses H3N2 with LOD of 13 viral particles in 1 μl, what is the lowest LOD for analysis of influenza viral particles by the glycan-based device achieved so far. The biosensor could detect H3N2 viruses selectively with a sensitivity ratio of 30 over influenza viruses H7N7. The impedimetric biosensor presented here is the most sensitive glycan-based device for detection of influenza viruses and among the most sensitive antibody or aptamer based biosensor devices.
- MeSH
- biosenzitivní techniky metody MeSH
- chřipka lidská diagnóza virologie MeSH
- impedanční spektroskopie metody MeSH
- lidé MeSH
- limita detekce MeSH
- polysacharidy chemie MeSH
- virus chřipky A, podtyp H3N2 izolace a purifikace MeSH
- virus chřipky A, podtyp H7N7 izolace a purifikace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
MicroRNAs (miRNAs) are small non-coding RNAs (18-22 nucleotides) that regulate gene expression and are associated with various diseases, including Laryngeal Cancer (LCa), which has a high mortality rate due to late diagnosis. Traditional methods for miRNA detection present several drawbacks (time-consuming steps, high cost and high false positive rate). Early-stage diagnosis and selective detection of miRNAs remain challenging. This study proposes a 3D flexible biosensor that combines nanofibers (NFs), gold nanoparticles (AuNPs), and an inverse molecular sentinel (iMS) for enzyme-free, SERS-based detection of miRNA-223-3p, evaluated as a potential LCa biomarker. The electrospun flexible nanofibers decorated with AuNPs enhance Raman signal. Selective detection of miRNA-223-3p is achieved by immobilizing an iMS-DNA probe labeled with a Raman reporter (Cyanine 3) on the AuNPs. The iMS distinctive stem-and-loop structure undergoes a conformational change upon interaction with the miRNA-223-3p, producing an "on to off" SERS signal. The proposed sensor demonstrated a linear detection range from 10 to 250 fM, with a limit of detection (LOD) of 19.50 ± 0.05 fM. The sensor selectivity was confirmed by analyzing the SERS signal behaviour in the presence of both Non-complementary miRNA and miRNA with three mismatched base pairs. This easily fabricable sensor requires no amplification and offers key advantages, including sensitivity, flexibility, and cost-effectiveness.
- MeSH
- biosenzitivní techniky * metody MeSH
- časná detekce nádoru metody MeSH
- kovové nanočástice * chemie MeSH
- lidé MeSH
- limita detekce MeSH
- mikro RNA * analýza genetika MeSH
- nádory hrtanu * diagnóza genetika MeSH
- nanovlákna * chemie MeSH
- Ramanova spektroskopie * metody MeSH
- zlato * chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The principle of synchronous detection (SD) has been applied to biosensor measurement. SD principle achieves significant increases in the signal to noise ratio, limit of detection and overall measurement robustness. Application of SD in biosensor measurement improves the analysis of the response and avoids the influence of interference/noise produced by stirring, electromagnetic effects and influence of parasitic currents. SD also enables the decomposition of signal to stimulation response and phenomena with long time of response. Second-order phenomena are identifiable in the signal. Linear statistical model was used to develop software for identification of the stimulation signal in the output current. SD was applied to the response signal of a Photosystem II complex (PSII) biosensor. PSII response to light stimulation follows first order kinetics. The inhibition kinetics of PSII has been studied. Kinetic constants of herbicide binding to PSII depend linearly on herbicide concentration and enable measurement of its concentration at low concentrations (linear range for diuron is 10⁻⁶ to 10⁻⁴ mM).
- MeSH
- biosenzitivní techniky metody MeSH
- časové faktory MeSH
- enzymy imobilizované antagonisté a inhibitory metabolismus MeSH
- fotosystém II (proteinový komplex) antagonisté a inhibitory metabolismus MeSH
- inhibitory enzymů farmakologie MeSH
- kinetika MeSH
- Synechococcus enzymologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
This paper reports a simple electrochemical strategy for the determination of microRNAs (miRNAs) using a commercial His-Tag-Zinc finger protein (His-Tag-ZFP) that binds preferably (but non-sequence specifically) RNA hybrids over ssRNAs, ssDNAs, and dsDNAs. The strategy involves the use of magnetic beads (His-Tag-Isolation-MBs) as solid support to capture the conjugate formed in homogenous solution between His-Tag-ZFP and the dsRNA homohybrid formed between the target miRNA (miR-21 selected as a model) and a biotinylated synthetic complementary RNA detector probe (b-RNA-Dp) further conjugated with a streptavidin-horseradish peroxidase (Strep-HRP) conjugate. The electrochemical detection is carried out by amperometry at disposable screen-printed carbon electrodes (SPCEs) (- 0.20 V vs Ag pseudo-reference electrode) upon magnetic capture of the resultant magnetic bioconjugates and H2O2 addition in the presence of hydroquinone (HQ). The as-prepared biosensor exhibits a dynamic concentration range from 3.0 to 100 nM and a detection limit (LOD) of 0.91 nM for miR-21 in just ~ 2 h. An acceptable discrimination was achieved between the target miRNA and other non-target nucleic acids (ssDNA, dsDNA, ssRNA, DNA-RNA, miR-122, miR-205, and single central- or terminal-base mismatched sequences). The biosensor was applied to the analysis of miR-21 from total RNA (RNAt) extracted from epithelial non-tumorigenic and adenocarcinoma breast cells without target amplification, pre-concentration, or reverse transcription steps. The versatility of the methodology due to the ZFP's non-sequence-specific binding behavior makes it easily extendable to determine any target RNA only by modifying the biotinylated detector probe.
The embedding microbial fuel cell (MFC) into constructed wetlands (CW) to form CW-MFC bears the potential to obtain bioelectricity and a clean environment. In this study, a bibliometric analysis using VOSviewer based on Web of Science data was conducted to provide an overview by tracing the development footprint of this technology. The countries, institutions, authors, key terms, and keywords were tracked and corresponding mapping was generated. From 2012 to September 2020, 442 authors from 129 organizations in 26 countries published 135 publications in 42 journals with total citation of 3139 times were found. The key terms analysis showed four clusters: bioelectricity generation performance, mechanism study, refractory pollutants removal, and enhanced conventional contaminants removal. Further research themes include exploring the biochemical properties of electrochemically active bacteria, emerging contaminants removal, effective bioelectricity harvest and the use, and biosensor development as well as scaling-up for real field application. The bibliometric results provide valuable references and information on potential research directions for future studies.
Affinity-based biosensing systems have become an important analytical tool for the detection and study of numerous biomolecules. The merging of these sensing technologies with microfluidic flow cells allows for faster detection times, increased sensitivities, and lower required sample volumes. In order to obtain a higher degree of performance from the sensor, it is important to know the effects of the flow cell geometry on the sensor sensitivity. In these sensors, the sensor sensitivity is related to the overall diffusive flux of analyte to the sensing surface; therefore increases in the analyte flux will be manifested as an increase in sensitivity, resulting in a lower limit of detection (LOD). Here we present a study pertaining to the effects of the flow cell height H on the analyte flux J, where for a common biosensor design we predict that the analyte flux will scale as J ≈ H(-2/3). We verify this scaling behavior via both numerical simulations as well as an experimental surface plasmon resonance (SPR) biosensor. We show the reduction of the flow cell height can have drastic effects on the sensor performance, where the LOD of our experimental system concerning the detection of ssDNA decreases by a factor of 4 when H is reduced from 47 μm to 7 μm. We utilize these results to discuss the applicability of this scaling behavior with respect to a generalized affinity-based biosensor.
Nanotechnology is one of the most promising future technologies for the food industry. Some of its applications have already been introduced in analytical techniques and food packaging technologies. This review summarizes existing knowledge about the implementation of nanotechnology in wine laboratory procedures. The focus is mainly on recent advancements in the design and development of nanomaterial-based sensors for wine compounds analysis and assessing wine safety. Nanotechnological approaches could be useful in the wine production process, to simplify wine analysis methods, and to improve the quality and safety of the final product.
- MeSH
- biosenzitivní techniky * MeSH
- nanostruktury * MeSH
- nanotechnologie MeSH
- obaly potravin MeSH
- víno * analýza MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Currently, the influenza virus infects millions of individuals every year. Since the influenza virus represents one of the greatest threats, it is necessary to develop a diagnostic technique that can quickly, inexpensively, and accurately detect the virus to effectively treat and control seasonal and pandemic strains. This study presents an alternative to current detection methods. The flow-injection analysis-based biosensor, which can rapidly and economically analyze a wide panel of influenza virus strains by using paramagnetic particles modified with glycan, can selectively bind to specific viral A/H5N1/Vietnam/1203/2004 protein-labeled quantum dots. Optimized detection of cadmium sulfide quantum dots (CdS QDs)-protein complexes connected to paramagnetic microbeads was performed using differential pulse voltammetry on the surface of a hanging mercury drop electrode (HMDE) and/or glassy carbon electrode (GCE). Detection limit (3 S/N) estimations based on cadmium(II) ions quantification were 0.1 μg/mL or 10 μg/mL viral protein at HMDE or GCE, respectively. Viral protein detection was directly determined using differential pulse voltammetry Brdicka reaction. The limit detection (3 S/N) of viral protein was estimated as 0.1 μg/mL. Streptavidin-modified paramagnetic particles were mixed with biotinylated selective glycan to modify their surfaces. Under optimized conditions (250 μg/mL of glycan, 30-min long interaction with viral protein, 25°C and 400 rpm), the viral protein labeled with quantum dots was selectively isolated and its cadmium(II) content was determined. Cadmium was present in detectable amounts of 10 ng per mg of protein. Using this method, submicrogram concentrations of viral proteins can be identified.
- MeSH
- biosenzitivní techniky přístrojové vybavení metody MeSH
- biotin chemie metabolismus MeSH
- elektrochemické techniky přístrojové vybavení metody MeSH
- elektrody MeSH
- hemaglutininové glykoproteiny viru chřipky analýza metabolismus MeSH
- kadmium analýza MeSH
- kvantové tečky MeSH
- limita detekce MeSH
- lineární modely MeSH
- magnetické nanočástice chemie MeSH
- průtoková injekční analýza přístrojové vybavení metody MeSH
- rtuť chemie MeSH
- sloučeniny kadmia chemie MeSH
- streptavidin chemie metabolismus MeSH
- sulfidy chemie MeSH
- uhlík chemie MeSH
- virus chřipky A, podtyp H5N1 chemie izolace a purifikace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A plasmonic biosensor for rapid detection of protein biomarkers in complex media is reported. Clinical serum samples were analyzed by using a novel biointerface architecture based on poly[(N-(2-hydroxypropyl) methacrylamide)-co-(carboxybetaine methacrylamide)] brushes functionalized with bioreceptors. This biointerface provided an excellent resistance to fouling even after the functionalization and allowed for the first time the direct detection of antibodies against hepatitis B surface antigen (anti-HBs) in clinical serum samples using surface plasmon resonance (SPR). The fabricated SPR biosensor allowed discrimination of anti-HBs positive and negative clinical samples in 10min. Results are validated by enzyme-linked immunoassays of the sera in a certified laboratory. The sensor could be regenerated by simple treatment with glycine buffer.
- MeSH
- akrylamidy chemie MeSH
- design vybavení MeSH
- hepatitida B - antigeny povrchové imunologie MeSH
- hepatitida B krev imunologie MeSH
- lidé MeSH
- limita detekce MeSH
- povrchová plasmonová rezonance přístrojové vybavení MeSH
- povrchové vlastnosti MeSH
- protilátky virové krev imunologie MeSH
- virus hepatitidy B imunologie izolace a purifikace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- validační studie MeSH
