Optimizing the Composition of the Substrate Enhances the Performance of Peroxidase-like Nanozymes in Colorimetric Assays: A Case Study of Prussian Blue and 3,3′-Diaminobenzidine
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Информация об архиве | Просмотр оригинала| Поле | Значение | |
| Заглавие |
Optimizing the Composition of the Substrate Enhances the Performance of Peroxidase-like Nanozymes in Colorimetric Assays: A Case Study of Prussian Blue and 3,3′-Diaminobenzidine
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| Автор |
Khramtsov, P.
Minin, A. Galaeva, Z. Mukhlynina, E. Kropaneva, M. Rayev, M. |
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| Тематика |
DOT BLOT
IMMUNOASSAY IMMUNOHISTOCHEMISTRY PEROXIDASE PRUSSIAN BLUE WESTERN BLOTTING COLORING AGENT DIAMINOBENZIDINE FERRIC FERROCYANIDE PEROXIDASE CATALYSIS CHEMISTRY COLORIMETRY PROCEDURES 3,3'-DIAMINOBENZIDINE CATALYSIS COLORIMETRY COLORING AGENTS PEROXIDASE PEROXIDASES |
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| Описание |
One of the emerging trends in modern analytical and bioanalytical chemistry involves the substitution of enzyme labels (such as horseradish peroxidase) with nanozymes (nanoparticles possessing enzyme-like catalytic activity). Since enzymes and nanozymes typically operate through different catalytic mechanisms, it is expected that optimal reaction conditions will also differ. The optimization of substrates for nanozymes usually focuses on determining the ideal pH and temperature. However, in some cases, even this step is overlooked, and commercial substrate formulations designed for enzymes are utilized. This paper demonstrates that not only the pH but also the composition of the substrate buffer, including the buffer species and additives, significantly impact the analytical signal generated by nanozymes. The presence of enhancers such as imidazole in commercial substrates diminishes the catalytic activity of nanozymes, which is demonstrated herein through the use of 3,3′-diaminobenzidine (DAB) and Prussian Blue as a model chromogenic substrate and nanozyme. Conversely, a simple modification to the substrate buffer greatly enhances the performance of nanozymes. Specifically, in this paper, it is demonstrated that buffers such as citrate, MES, HEPES, and TRIS, containing 1.5–2 M NaCl or NH4Cl, substantially increase DAB oxidation by Prussian Blue and yield a higher signal compared to commercial DAB formulations. The central message of this paper is that the optimization of substrate composition should be an integral step in the development of nanozyme-based assays. Herein, a step-by-step optimization of the DAB substrate composition for Prussian Blue nanozymes is presented. The optimized substrate outperforms commercial formulations in terms of efficiency. The effectiveness of the optimized DAB substrate is affirmed through its application in several commonly used immunostaining techniques, including tissue staining, Western blotting assays of immunoglobulins, and dot blot assays of antibodies against SARS-CoV-2. © 2023 by the authors.
Russian Science Foundation, RSF: 22-24-20091 This study was supported by the Russian Science Foundation and the Government of Perm krai, grant 22-24-20091. |
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| Дата |
2024-04-05T16:36:51Z
2024-04-05T16:36:51Z 2023 |
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| Тип |
Article
Journal article (info:eu-repo/semantics/article) |info:eu-repo/semantics/publishedVersion |
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| Идентификатор |
Khramtsov, P, Minin, A, Galaeva, Z, Mukhlynina, E, Kropaneva, M & Rayev, M 2023, 'Optimizing the Composition of the Substrate Enhances the Performance of Peroxidase-like Nanozymes in Colorimetric Assays: A Case Study of Prussian Blue and 3,3′-Diaminobenzidine', Molecules, Том. 28, № 22, 7622. https://doi.org/10.3390/molecules28227622
Khramtsov, P., Minin, A., Galaeva, Z., Mukhlynina, E., Kropaneva, M., & Rayev, M. (2023). Optimizing the Composition of the Substrate Enhances the Performance of Peroxidase-like Nanozymes in Colorimetric Assays: A Case Study of Prussian Blue and 3,3′-Diaminobenzidine. Molecules, 28(22), [7622]. https://doi.org/10.3390/molecules28227622 1420-3049 Final All Open Access, Gold, Green https://www.scopus.com/inward/record.uri?eid=2-s2.0-85177784935&doi=10.3390%2fmolecules28227622&partnerID=40&md5=367cb7231c7c94b525006bb036005002 https://www.mdpi.com/1420-3049/28/22/7622/pdf?version=1700123277 http://elar.urfu.ru/handle/10995/130991 10.3390/molecules28227622 85177784935 001113971200001 |
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| Язык |
en
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| Связанные ресурсы |
info:eu-repo/grantAgreement/RSF//22-24-20091
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| Права |
Open access (info:eu-repo/semantics/openAccess)
cc-by https://creativecommons.org/licenses/by/4.0/ |
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| Формат |
application/pdf
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| Издатель |
Multidisciplinary Digital Publishing Institute (MDPI)
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| Источник |
Molecules
Molecules |
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