Sensitive Silver-Enhanced Microplate Apta-Enzyme Assay of Sb<sup>3+</sup> Ions in Drinking and Natural Waters

The toxic effects of antimony pose risks to human health. Therefore, simple analytical techniques for its widescale monitoring in water sources are in demand. In this study, a sensitive microplate apta-enzyme assay for Sb<sup>3+</sup> detection was developed. The biotinylated aptamer A&l...

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Main Authors: Nadezhda S. Komova, Kseniya V. Serebrennikova, Anna N. Berlina, Anatoly V. Zherdev, Boris B. Dzantiev
Format: Article
Language:English
Published: MDPI AG 2023-10-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/28/19/6973
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author Nadezhda S. Komova
Kseniya V. Serebrennikova
Anna N. Berlina
Anatoly V. Zherdev
Boris B. Dzantiev
author_facet Nadezhda S. Komova
Kseniya V. Serebrennikova
Anna N. Berlina
Anatoly V. Zherdev
Boris B. Dzantiev
author_sort Nadezhda S. Komova
collection DOAJ
description The toxic effects of antimony pose risks to human health. Therefore, simple analytical techniques for its widescale monitoring in water sources are in demand. In this study, a sensitive microplate apta-enzyme assay for Sb<sup>3+</sup> detection was developed. The biotinylated aptamer A<sub>10</sub> was hybridized with its complementary biotinylated oligonucleotide T<sub>10</sub> and then immobilized on the surface of polysterene microplate wells. Streptavidin labeled with horseradish peroxidase (HRP) bound to the biotin of a complementary complex and transformed the 3,3′,5,5′-tetramethylbenzidine substrate, generating an optical signal. Sb<sup>3+</sup> presenting in the sample bounded to an A<sub>10</sub> aptamer, thus releasing T<sub>10</sub>, preventing streptavidin-HRP binding and, as a result, reducing the optical signal. This effect allowed for the detection of Sb<sup>3+</sup> with a working range from 0.09 to 2.3 µg/mL and detection limit of 42 ng/mL. It was established that the presence of Ag<sup>+</sup> at the stage of A<sub>10</sub>/T<sub>10</sub> complex formation promoted dehybridization of the aptamer A<sub>10</sub> and the formation of the A<sub>10</sub>/Sb<sup>3+</sup> complex. The working range of the Ag<sup>+</sup>-enhanced microplate apta-enzyme assay for Sb<sup>3+</sup> was determined to be 8–135 ng/mL, with a detection limit of 1.9 ng/mL. The proposed enhanced approach demonstrated excellent selectivity against other cations/anions, and its practical applicability was confirmed through an analysis of drinking and spring water samples with recoveries of Sb<sup>3+</sup> in the range of 109.0–126.2% and 99.6–106.1%, respectively.
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spelling doaj.art-3f0982b29d0e4225a3f69dea6102ef122023-11-19T14:48:03ZengMDPI AGMolecules1420-30492023-10-012819697310.3390/molecules28196973Sensitive Silver-Enhanced Microplate Apta-Enzyme Assay of Sb<sup>3+</sup> Ions in Drinking and Natural WatersNadezhda S. Komova0Kseniya V. Serebrennikova1Anna N. Berlina2Anatoly V. Zherdev3Boris B. Dzantiev4A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, RussiaA.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, RussiaA.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, RussiaA.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, RussiaA.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, 119071 Moscow, RussiaThe toxic effects of antimony pose risks to human health. Therefore, simple analytical techniques for its widescale monitoring in water sources are in demand. In this study, a sensitive microplate apta-enzyme assay for Sb<sup>3+</sup> detection was developed. The biotinylated aptamer A<sub>10</sub> was hybridized with its complementary biotinylated oligonucleotide T<sub>10</sub> and then immobilized on the surface of polysterene microplate wells. Streptavidin labeled with horseradish peroxidase (HRP) bound to the biotin of a complementary complex and transformed the 3,3′,5,5′-tetramethylbenzidine substrate, generating an optical signal. Sb<sup>3+</sup> presenting in the sample bounded to an A<sub>10</sub> aptamer, thus releasing T<sub>10</sub>, preventing streptavidin-HRP binding and, as a result, reducing the optical signal. This effect allowed for the detection of Sb<sup>3+</sup> with a working range from 0.09 to 2.3 µg/mL and detection limit of 42 ng/mL. It was established that the presence of Ag<sup>+</sup> at the stage of A<sub>10</sub>/T<sub>10</sub> complex formation promoted dehybridization of the aptamer A<sub>10</sub> and the formation of the A<sub>10</sub>/Sb<sup>3+</sup> complex. The working range of the Ag<sup>+</sup>-enhanced microplate apta-enzyme assay for Sb<sup>3+</sup> was determined to be 8–135 ng/mL, with a detection limit of 1.9 ng/mL. The proposed enhanced approach demonstrated excellent selectivity against other cations/anions, and its practical applicability was confirmed through an analysis of drinking and spring water samples with recoveries of Sb<sup>3+</sup> in the range of 109.0–126.2% and 99.6–106.1%, respectively.https://www.mdpi.com/1420-3049/28/19/6973antimony ionsmicroplate apta-enzyme assayaptameroligonucleotideadeninethymine
spellingShingle Nadezhda S. Komova
Kseniya V. Serebrennikova
Anna N. Berlina
Anatoly V. Zherdev
Boris B. Dzantiev
Sensitive Silver-Enhanced Microplate Apta-Enzyme Assay of Sb<sup>3+</sup> Ions in Drinking and Natural Waters
Molecules
antimony ions
microplate apta-enzyme assay
aptamer
oligonucleotide
adenine
thymine
title Sensitive Silver-Enhanced Microplate Apta-Enzyme Assay of Sb<sup>3+</sup> Ions in Drinking and Natural Waters
title_full Sensitive Silver-Enhanced Microplate Apta-Enzyme Assay of Sb<sup>3+</sup> Ions in Drinking and Natural Waters
title_fullStr Sensitive Silver-Enhanced Microplate Apta-Enzyme Assay of Sb<sup>3+</sup> Ions in Drinking and Natural Waters
title_full_unstemmed Sensitive Silver-Enhanced Microplate Apta-Enzyme Assay of Sb<sup>3+</sup> Ions in Drinking and Natural Waters
title_short Sensitive Silver-Enhanced Microplate Apta-Enzyme Assay of Sb<sup>3+</sup> Ions in Drinking and Natural Waters
title_sort sensitive silver enhanced microplate apta enzyme assay of sb sup 3 sup ions in drinking and natural waters
topic antimony ions
microplate apta-enzyme assay
aptamer
oligonucleotide
adenine
thymine
url https://www.mdpi.com/1420-3049/28/19/6973
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