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Flow-Through Acetylcholinesterase Sensor with Replaceable Enzyme Reactor

Fast and reliable determination of enzyme inhibitors are of great importance in environmental monitoring and biomedicine because of the high biological activity and toxicity of such species and the necessity of their reliable assessment in many media. In this work, a flow-through biosensor has been...

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Autores principales: Ivanov, Alexey, Stoikov, Dmitry, Shafigullina, Insiya, Shurpik, Dmitry, Stoikov, Ivan, Evtugyn, Gennady
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9496324/
https://www.ncbi.nlm.nih.gov/pubmed/36140061
http://dx.doi.org/10.3390/bios12090676
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author Ivanov, Alexey
Stoikov, Dmitry
Shafigullina, Insiya
Shurpik, Dmitry
Stoikov, Ivan
Evtugyn, Gennady
author_facet Ivanov, Alexey
Stoikov, Dmitry
Shafigullina, Insiya
Shurpik, Dmitry
Stoikov, Ivan
Evtugyn, Gennady
author_sort Ivanov, Alexey
collection PubMed
description Fast and reliable determination of enzyme inhibitors are of great importance in environmental monitoring and biomedicine because of the high biological activity and toxicity of such species and the necessity of their reliable assessment in many media. In this work, a flow-through biosensor has been developed and produced by 3D printing from poly(lactic acid). Acetylcholinesterase from an electric eel was immobilized on the inner walls of the reactor cell. The concentration of thiocholine formed in enzymatic hydrolysis of the substrate was monitored amperometrically with a screen-printed carbon electrode modified with carbon black particles, pillar[5]arene, electropolymerized Methylene blue and thionine. In the presence of thiocholine, the cathodic current at −0.25 V decreased because of an alternative chemical reaction of the macrocycle. The conditions of enzyme immobilization and signal measurements were optimized and the performance of the biosensor was assessed in the determination of reversible (donepezil, berberine) and irreversible (carbofuran) inhibitors. In the optimal conditions, the flow-through biosensor made it possible to determine 1.0 nM–1.0 μM donepezil, 1.0 μM–1.0 mM berberine and 10 nM to 0.1 μM carbofuran. The AChE biosensor was tested on spiked samples of artificial urine for drugs and peanuts for carbofuran. Possible interference of the sample components was eliminated by dilution of the samples with phosphate buffer. Easy mounting, low cost of replaceable parts of the cell and satisfactory analytical and metrological characteristics made the biosensor a promising future application as a point-of-care or point-of-demand device outside of a chemical laboratory.
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spelling pubmed-94963242022-09-23 Flow-Through Acetylcholinesterase Sensor with Replaceable Enzyme Reactor Ivanov, Alexey Stoikov, Dmitry Shafigullina, Insiya Shurpik, Dmitry Stoikov, Ivan Evtugyn, Gennady Biosensors (Basel) Article Fast and reliable determination of enzyme inhibitors are of great importance in environmental monitoring and biomedicine because of the high biological activity and toxicity of such species and the necessity of their reliable assessment in many media. In this work, a flow-through biosensor has been developed and produced by 3D printing from poly(lactic acid). Acetylcholinesterase from an electric eel was immobilized on the inner walls of the reactor cell. The concentration of thiocholine formed in enzymatic hydrolysis of the substrate was monitored amperometrically with a screen-printed carbon electrode modified with carbon black particles, pillar[5]arene, electropolymerized Methylene blue and thionine. In the presence of thiocholine, the cathodic current at −0.25 V decreased because of an alternative chemical reaction of the macrocycle. The conditions of enzyme immobilization and signal measurements were optimized and the performance of the biosensor was assessed in the determination of reversible (donepezil, berberine) and irreversible (carbofuran) inhibitors. In the optimal conditions, the flow-through biosensor made it possible to determine 1.0 nM–1.0 μM donepezil, 1.0 μM–1.0 mM berberine and 10 nM to 0.1 μM carbofuran. The AChE biosensor was tested on spiked samples of artificial urine for drugs and peanuts for carbofuran. Possible interference of the sample components was eliminated by dilution of the samples with phosphate buffer. Easy mounting, low cost of replaceable parts of the cell and satisfactory analytical and metrological characteristics made the biosensor a promising future application as a point-of-care or point-of-demand device outside of a chemical laboratory. MDPI 2022-08-24 /pmc/articles/PMC9496324/ /pubmed/36140061 http://dx.doi.org/10.3390/bios12090676 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ivanov, Alexey
Stoikov, Dmitry
Shafigullina, Insiya
Shurpik, Dmitry
Stoikov, Ivan
Evtugyn, Gennady
Flow-Through Acetylcholinesterase Sensor with Replaceable Enzyme Reactor
title Flow-Through Acetylcholinesterase Sensor with Replaceable Enzyme Reactor
title_full Flow-Through Acetylcholinesterase Sensor with Replaceable Enzyme Reactor
title_fullStr Flow-Through Acetylcholinesterase Sensor with Replaceable Enzyme Reactor
title_full_unstemmed Flow-Through Acetylcholinesterase Sensor with Replaceable Enzyme Reactor
title_short Flow-Through Acetylcholinesterase Sensor with Replaceable Enzyme Reactor
title_sort flow-through acetylcholinesterase sensor with replaceable enzyme reactor
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9496324/
https://www.ncbi.nlm.nih.gov/pubmed/36140061
http://dx.doi.org/10.3390/bios12090676
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