Browse
Submit Data
Databases
API
Help

Dataset Information

25 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Sensitive Identification of Microcystin-LR via a Reagent-Free and Reusable Electrochemical Biosensor Using a Methylene Blue-Labeled Aptamer.

ABSTRACT: We report a methylene blue (MB)-modified electrochemical aptamer (E-AB) sensor for determining microcystin-LR (MC-LR). The signal transduction of the sensor was based on changes in conformation and position of MB induced by the binding between MC-LR and the modified aptamer probe. In the absence of MC-LR, an aptamer probe was considered partially folded. After combining aptamer and MC-LR, the configuration of the aptamer probe changed and facilitated the electron transfer between MB and the electrode surface. As a result, an increased current response was observed. We optimized the parameters and evaluated the electrochemical performance of the sensor using square wave voltammetry (SWV). MC-LR was measured from 1.0 to 750.0 ng/L with a detection limit of 0.53 ng/L. The reliability of the method was verified by the determination of MC-LR in environmental real samples, such as pond water and tap water. Moreover, we demonstrated that this reagent-less biosensor could be regenerated and reused after rinsing with deionized water with good accuracy and reproducibility. As a reusable and regenerable E-AB sensor, this rapid, reagent-free, and sensitive sensing platform will facilitate routine monitoring of MC-LR in actual samples.

SUBMITTER: Wei X

PROVIDER: S-EPMC9332554 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Json Xml

Similar Datasets

Detecting Redox Potentials Using Porous Boron Nitride/ATP-DNA Aptamer/Methylene Blue Biosensor to Monitor Microbial Activities.

Project description:Microbial activity has gained attention because of its impact on the environment and the quality of people's lives. Most of today's methods, which include genome sequencing and electrochemistry, are costly and difficult to manage. Our group proposed a method using the redox potential change to detect microbial activity, which is rooted in the concept that metabolic activity can change the redox potential of a microbial community. The redox potential change was captured by a biosensor consisting of porous boron nitride, ATP-DNA aptamer, and methylene blue as the fluorophore. This assembly can switch on or off when there is a redox potential change, and this change leads to a fluorescence change that can be examined using a multipurpose microplate reader. The results show that this biosensor can detect microbial community changes when its composition is changed or toxic metals are ingested.

| S-EPMC8777636 | biostudies-literature

Aptamer-based electrochemical biosensor for interferon gamma detection.

Project description:In this paper, we describe the development of an electrochemical DNA aptamer-based biosensor for detection of interferon (IFN)-?. A DNA hairpin containing IFN-?-binding aptamer was thiolated, conjugated with methylene blue (MB) redox tag, and immobilized on a gold electrode by self-assembly. Binding of IFN-? caused the aptamer hairpin to unfold, pushing MB redox molecules away from the electrode and decreasing electron-transfer efficiency. The change in redox current was quantified using square wave voltammetry (SWV) and was found to be highly sensitive to IFN-? concentration. The limit of detection for optimized biosensor was 0.06 nM with linear response extending to 10 nM. This aptasensor was specific to IFN-? in the presence of overabundant serum proteins. Importantly, the same aptasensor could be regenerated by disrupting aptamer-IFN-? complex in urea buffer and reused multiple times. Unlike standard sandwich immunoassays, the aptasensor described here allowed one to detect IFN-? binding directly without the need for multiple washing steps and reagents. An electrochemical biosensor for simple and sensitive detection of IFN-? demonstrated in this paper will have future applications in immunology, cancer research, and infectious disease monitoring.

| S-EPMC2948235 | biostudies-literature

Microcystin-LR (MC-LR) Triggers Inflammatory Responses in Macrophages.

Project description:We were the first to previously report that microcystin-LR (MC-LR) has limited effects within the colons of healthy mice but has toxic effects within colons of mice with pre-existing inflammatory bowel disease. In the current investigation, we aimed to elucidate the mechanism by which MC-LR exacerbates colitis and to identify effective therapeutic targets. Through our current investigation, we report that there is a significantly greater recruitment of macrophages into colonic tissue with pre-existing colitis in the presence of MC-LR than in the absence of MC-LR. This is seen quantitatively through IHC staining and the enumeration of F4/80-positive macrophages and through gene expression analysis for Cd68, Cd11b, and Cd163. Exposure of isolated macrophages to MC-LR was found to directly upregulate macrophage activation markers Tnf and Il1b. Through a high-throughput, unbiased kinase activity profiling strategy, MC-LR-induced phosphorylation events were compared with potential inhibitors, and doramapimod was found to effectively prevent MC-LR-induced inflammatory responses in macrophages.

| S-EPMC8472269 | biostudies-literature

Supramolecular Enzymatic Labeling for Aptamer Switch-Based Electrochemical Biosensor.

Project description:Here we report a novel labeling strategy for electrochemical aptasensors based on enzymatic marking via supramolecular host-guest interactions. This approach relies on the use of an adamantane-modified target-responsive hairpin DNA aptamer as an affinity bioreceptor, and a neoglycoconjugate of β-cyclodextin (CD) covalently attached to a redox enzyme as a labeling element. As a proof of concept, an amperometric aptasensor for a carcinoembryonic antigen was assembled on screen-printed carbon electrodes modified with electrodeposited fern-like gold nanoparticles/graphene oxide and, by using a horseradish peroxidase-CD neoglycoenzyme as a biocatalytic redox label. This aptasensor was able to detect the biomarker in the concentration range from 10 pg/mL to 1 ng/mL with a high selectivity and a low detection limit of 3.1 pg/mL in human serum samples.

| S-EPMC9313153 | biostudies-literature

Microarray Analysis of Microcystin-LR

2014-07-25 | GSE59495 | GEO

Electrochemical Flow-ELISA for Rapid and Sensitive Determination of Microcystin-LR Using Automated Sequential Injection System.

Project description:An amperometric immunoanalysis system based on monoclonal antibodies immobilized on Sepharose beads and packed into a micro-immunocolumn was developed for the quantification of microcystin-LR. Microcystin-LR (MCLR) was used as a reference microcystin variant. Inside the immunocolumn, free microcystins and microcystin-horseradish peroxidase (tracer) were sequentially captured by the immobilized antibodies, and the detection was performed electrochemically using Super AquaBlue ELISA substrate 2,2'-azinobis(3-ethylbenzothiazoline-sulfonic acid) (ABTS). The ABTS●+ generated by enzymatic oxidation of ABTS was electrochemically determined at a carbon working electrode by applying a reduction potential set at 0.4 V versus Ag/AgCl reference electrode. The peak current intensity was inversely proportional to the amount of analyte bound to the immunocolumn. The amperometric flow-ELISA system, which was automatically controlled through the CapSenzeTM (Lund, Sweden) computer software, enabled determination of MCLR as low as 0.01 µg/L. The assay time was very short (20 min for one assay cycle). In addition, the electrochemical signals were not significantly affected by possible interferences which could be present in the real samples. Along with the simplicity of automation, this makes the developed method a promising tool for use in water quality assessment.

| S-EPMC5539796 | biostudies-other

Microarray Analysis of Microcystin-LR

Project description:Microcystins are produced by the cyanobacteria, most commonly Microcystis aerµginosa. Upon ingestion, toxic microcystins are actively absorbed by fish, birds and mammals where they are primarily liver toxins. Groups of 4-6 rats were exposed to 0, 1, 10, 50 or 100 µg/kg microcystin-LR for 0.5, 1, 3 or 6 hours and gene expression analysis performed on liver with samples hybridized to whole rat genome RG230_2.0 GeneChip arrays (Affymetrix, CA).

2014-07-25 | E-GEOD-59495 | biostudies-arrayexpress

Poly(methylene blue)-Based Electrochemical Platform for Label-Free Sensing of Acrylamide.

Project description:The present work reports the electrochemical sensing of acrylamide (AM) using a poly(methylene blue)-modified glassy carbon electrode (PMB/GCE) where PMB functions as an electrochemical reporter. PMB was prepared by electrochemical polymerization of methylene blue. Electrochemical sensing of AM was facilitated by the interaction between AM and PMB. Further the interaction between AM and PMB was investigated using ultraviolet-visible (UV-vis) spectroscopy and Raman analysis. The surface morphology was confirmed by atomic force microscopy (AFM) and field emission scanning electron microscopy (FESEM) analyses. PMB/GCE was further characterized by X-ray photoelectron spectroscopy (XPS), and the electrochemical performance was assessed using cyclic voltammetry and differential pulse voltammetry. Cyclic voltammetry analysis showed a decrease in current at the redox center of PMB upon addition of AM. The association constant and binding number of AM with PMB/GCE were calculated using differential pulse voltammetry and found to be 8.9 × 106 M-1 and 0.64 (∼1), respectively. The results indicated a strong interaction of AM on the PMB/GCE surface. Further, chronocoulometry analysis of PMB/GCE in the presence of AM showed a decrease in charge due to the interaction of AM with PMB. Under optimized conditions, PMB/GCE exhibited a decrease in current proportional to the concentration of AM in the range of 0.025-16 μM with sensitivity and detection limit 0.252 μA nM-1 and 0.13 nM, respectively. Real sample analysis was carried out by the standard addition method using the solution extracted from potato chips.

| S-EPMC8047665 | biostudies-literature

Selection of high affinity aptamer-ligand for dexamethasone and its electrochemical biosensor.

Project description:A high specificity aptamer-ligand biorecognition and binding system to monitor of dexamethasone (DXN) was developed. The detection principle was based on a label-free electrochemical aptasensor. The selection of the aptamer was successfully performed by the systematic evolution of ligands through exponential enrichment technique (SELEX). From a random library of 1.08?×?1015 single-stranded DNA, an aptamer designated as DEX04 showed a highest affinity with a dissociation constant of 18.35?nM. It also showed a good conformational change when binding with DXN. In addition, the aptamer DEX04 did not show any cross-reactivity with other commonly used hormones. An impedimetric aptasensor for DXN was then developed by immobilizing DEX04 on a gold electrode. The binding upon to DXN was monitored by following the change in the charge transfer resistance (Rct) of the [Fe(CN)6]4-/3- redox couple. The aptasensor exhibited a linear range from 2.5 to 100?nM with a detection limit of 2.12?nM. When applied aptasensor to test in water samples, it showed good recovery percentages. The new DXN aptamer can be employed in other biosensing applications for food control and the diagnosis of some diseases in medicine as a cost-effective, sensitive and rapid detection method.

| S-EPMC6488579 | biostudies-literature

Aptamer-Based Microfluidic Electrochemical Biosensor for Monitoring Cell-Secreted Trace Cardiac Biomarkers.

Project description:Continual monitoring of secreted biomarkers from organ-on-a-chip models is desired to understand their responses to drug exposure in a noninvasive manner. To achieve this goal, analytical methods capable of monitoring trace amounts of secreted biomarkers are of particular interest. However, a majority of existing biosensing techniques suffer from limited sensitivity, selectivity, stability, and require large working volumes, especially when cell culture medium is involved, which usually contains a plethora of nonspecific binding proteins and interfering compounds. Hence, novel analytical platforms are needed to provide noninvasive, accurate information on the status of organoids at low working volumes. Here, we report a novel microfluidic aptamer-based electrochemical biosensing platform for monitoring damage to cardiac organoids. The system is scalable, low-cost, and compatible with microfluidic platforms easing its integration with microfluidic bioreactors. To create the creatine kinase (CK)-MB biosensor, the microelectrode was functionalized with aptamers that are specific to CK-MB biomarker secreted from a damaged cardiac tissue. Compared to antibody-based sensors, the proposed aptamer-based system was highly sensitive, selective, and stable. The performance of the sensors was assessed using a heart-on-a-chip system constructed from human embryonic stem cell-derived cardiomyocytes following exposure to a cardiotoxic drug, doxorubicin. The aptamer-based biosensor was capable of measuring trace amounts of CK-MB secreted by the cardiac organoids upon drug treatments in a dose-dependent manner, which was in agreement with the beating behavior and cell viability analyses. We believe that, our microfluidic electrochemical biosensor using aptamer-based capture mechanism will find widespread applications in integration with organ-on-a-chip platforms for in situ detection of biomarkers at low abundance and high sensitivity.

| S-EPMC5844853 | biostudies-literature

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data