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
Abstract
:1. Introduction
2. Results and Discussion
2.1. Characterization of Nanoparticles
2.2. DAB Substrate Optimization
- A 33 mM Na-citrate with 1.5 M NaCl, pH 6, 1 mg/mL DAB; 0.1% H2O2.
- A 33 mM Na-citrate with 1.5 M NH4Cl, pH 7, 1 mg/mL DAB; 0.1% H2O2;
- A 30 mM HEPES–HCl, 3 mM Na-citrate, with 1.5 M NaCl, pH 7, 1 mg/mL DAB; 0.1% H2O2.
2.3. The Effect of Substrate Buffer in Nanozyme-Based Immunostaining
3. Materials and Methods
3.1. Materials
3.2. Synthesis of Prussian Blue Nanoparticles
3.3. Conjugation of Prussian Blue Nanoparticles with Streptavidin
3.4. Characterization of Nanoparticles
3.5. DAB Substrate Optimization
3.6. Direct Assay for Bi-BSA Detection with PB@GelA/Str and PB@GelA/BSA
3.7. Direct Assay for Bi-BSA Detection with HRP-Str
3.8. Western Blotting
3.9. Dot Blot Assay of Antibodies against Spike Protein of SARS-CoV-2
3.10. Immunohistochemistry
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Scrimin, P.; Prins, L.J. Sensing through Signal Amplification. Chem. Soc. Rev. 2011, 40, 4488–4505. [Google Scholar] [CrossRef] [PubMed]
- Krainer, F.W.; Glieder, A. An Updated View on Horseradish Peroxidases: Recombinant Production and Biotechnological Applications. Appl. Microbiol. Biotechnol. 2015, 99, 1611–1625. [Google Scholar] [CrossRef] [PubMed]
- Wang, Q.; Wei, H.; Zhang, Z.; Wang, E.; Dong, S. Nanozyme: An Emerging Alternative to Natural Enzyme for Biosensing and Immunoassay. TrAC Trends Anal. Chem. 2018, 105, 218–224. [Google Scholar] [CrossRef]
- Chi, Z.; Wang, Q.; Gu, J. Recent Advances in Colorimetric Sensors Based on Nanozymes with Peroxidase-like Activity. Analyst 2023, 148, 487–506. [Google Scholar] [CrossRef] [PubMed]
- Zandieh, M.; Liu, J. Nanozymes: Definition, Activity, and Mechanisms. Adv. Mater. 2023, 35, e2211041. [Google Scholar] [CrossRef] [PubMed]
- Shen, X.; Wang, Z.; Gao, X.J.; Gao, X. Reaction Mechanisms and Kinetics of Nanozymes: Insights from Theory and Computation. Adv. Mater. 2023, 35, e2211151. [Google Scholar] [CrossRef]
- Rimm, D.L. What Brown Cannot Do for You. Nat. Biotechnol. 2006, 24, 914–916. [Google Scholar] [CrossRef] [PubMed]
- Kalyuzhny, A.E. Chemistry and Biology of the ELISPOT Assay. In Handbook of ELISPOT. Methods in Molecular Biology; Humana Press: Totowa, NJ, USA, 2005; Volume 302, pp. 15–31. [Google Scholar] [CrossRef]
- Gupta, B.; Yang, G.; Petrauskene, O.; Key, M. Recent Advances in Chromogens for Immunohistochemistry. In Signal Transduction Immunohistochemistry. Methods in Molecular Biology; Humana Press: New York, NY, USA, 2023; Volume 2593, pp. 35–50. [Google Scholar] [CrossRef]
- Alegria-Schaffer, A.; Lodge, A.; Vattem, K. Performing and Optimizing Western Blots with an Emphasis on Chemiluminescent Detection. Methods Enzymol. 2009, 463, 573–599. [Google Scholar] [CrossRef] [PubMed]
- Calabria, D.; Calabretta, M.M.; Zangheri, M.; Marchegiani, E.; Trozzi, I.; Guardigli, M.; Michelini, E.; Di Nardo, F.; Anfossi, L.; Baggiani, C.; et al. Recent Advancements in Enzyme-Based Lateral Flow Immunoassays. Sensors 2021, 21, 3358. [Google Scholar] [CrossRef]
- Heras, A.; Key, M. Stable Substrate-Chromogen Solutions for Enenzyme Activity Detection. U.S. Patent 5804404A, 8 September 1998. [Google Scholar]
- Kasamatsu, T.; Kitano, Y. Dab-Containing Substrate Kit for Dyeing Use Which Is Produced Using Labelling Enzyme. U.S. Patent 20160010140A1, 14 January 2016. [Google Scholar]
- Straus, W. Imidazole Increases the Sensitivity of the Cytochemical Reaction for Peroxidase with Diaminobenzidine at a Neutral pH. J. Histochem. Cytochem. 1982, 30, 491–493. [Google Scholar] [CrossRef]
- Greenfield, L.; Starkenburg, S.; Shallice, M.; Nyhus, J.; Leong, L. Stable Compositions Comprising Chromogenic Compounds and Methods of Use. U.S. Patent US20120077211A1, 15 December 2009. [Google Scholar]
- Hsu, S.M.; Soban, E. Color Modification of Diaminobenzidine (DAB) Precipitation by Metallic Ions and Its Application for Double Immunohistochemistry. J. Histochem. Cytochem. 1982, 30, 1079–1082. [Google Scholar] [CrossRef]
- Zhang, W.; Dong, J.; Wu, Y.; Cao, P.; Song, L.; Ma, M.; Gu, N.; Zhang, Y. Shape-Dependent Enzyme-like Activity of Co3O4 Nanoparticles and Their Conjugation with His-Tagged EGFR Single-Domain Antibody. Colloids Surf. B Biointerfaces 2017, 154, 55–62. [Google Scholar] [CrossRef]
- Dong, J.; Song, L.; Yin, J.-J.; He, W.; Wu, Y.; Gu, N.; Zhang, Y. Co3O4 Nanoparticles with Multi-Enzyme Activities and Their Application in Immunohistochemical Assay. ACS Appl. Mater. Interfaces 2014, 6, 1959–1970. [Google Scholar] [CrossRef] [PubMed]
- Wu, Y.; Song, M.; Xin, Z.; Zhang, X.; Zhang, Y.; Wang, C.; Li, S.; Gu, N. Ultra-Small Particles of Iron Oxide as Peroxidase for Immunohistochemical Detection. Nanotechnology 2011, 22, 225703. [Google Scholar] [CrossRef] [PubMed]
- Jiang, B.; Yan, L.; Zhang, J.; Zhou, M.; Shi, G.; Tian, X.; Fan, K.; Hao, C.; Yan, X. Biomineralization Synthesis of the Cobalt Nanozyme in SP94-Ferritin Nanocages for Prognostic Diagnosis of Hepatocellular Carcinoma. ACS Appl. Mater. Interfaces 2019, 11, 9747–9755. [Google Scholar] [CrossRef]
- Hormozi Jangi, S.R.; Akhond, M.; Absalan, G. A Field-Applicable Colorimetric Assay for Notorious Explosive Triacetone Triperoxide through Nanozyme-Catalyzed Irreversible Oxidation of 3, 3′-Diaminobenzidine. Microchim. Acta 2020, 187, 431. [Google Scholar] [CrossRef]
- Moreno-Castilla, C.; Naranjo, Á.; Victoria López-Ramón, M.; Siles, E.; López-Peñalver, J.J.; de Almodóvar, J.M.R. Influence of the Hydrodynamic Size and ζ Potential of Manganese Ferrite Nanozymes as Peroxidase-Mimicking Catalysts at pH 4 in Different Buffers. J. Catal. 2022, 414, 179–185. [Google Scholar] [CrossRef]
- Filippova, A.D.; Sozarukova, M.M.; Baranchikov, A.E.; Kottsov, S.Y.; Cherednichenko, K.A.; Ivanov, V.K. Peroxidase-like Activity of CeO2 Nanozymes: Particle Size and Chemical Environment Matter. Molecules 2023, 28, 3811. [Google Scholar] [CrossRef]
- Du, Z.; Zhu, L.; Wang, P.; Lan, X.; Lin, S.; Xu, W. Coordination-Driven One-Step Rapid Self-Assembly Synthesis of Dual-Functional Ag@Pt Nanozyme. Small Weinh. Bergstr. Ger. 2023, 19, e2301048. [Google Scholar] [CrossRef]
- Wu, Y.; Yang, L.; Wu, Q.; Liu, Q.; Zou, L.; Yang, X.; Tang, K. Regulation of the Oxidase Mimetic Activity of Ceria Nanoparticles by Buffer Composition. Chem. Weinh. Bergstr. Ger. 2023, 29, e202204071. [Google Scholar] [CrossRef]
- Sun, J.; Li, C.; Qi, Y.; Guo, S.; Liang, X. Optimizing Colorimetric Assay Based on V2O5 Nanozymes for Sensitive Detection of H2O2 and Glucose. Sensors 2016, 16, 584. [Google Scholar] [CrossRef]
- Tian, S.; Zhang, C.; Yu, M.; Li, Y.; Fan, L.; Li, X. Buffer Species-Dependent Catalytic Activity of Cu-Adenine as a Laccase Mimic for Constructing Sensor Array to Identify Multiple Phenols. Anal. Chim. Acta 2022, 1204, 339725. [Google Scholar] [CrossRef]
- Komkova, M.A.; Karyakina, E.E.; Karyakin, A.A. Catalytically Synthesized Prussian Blue Nanoparticles Defeating Natural Enzyme Peroxidase. J. Am. Chem. Soc. 2018, 140, 11302–11307. [Google Scholar] [CrossRef]
- Khramtsov, P.; Kropaneva, M.; Minin, A.; Bochkova, M.; Timganova, V.; Maximov, A.; Puzik, A.; Zamorina, S.; Rayev, M. Prussian Blue Nanozymes with Enhanced Catalytic Activity: Size Tuning and Application in ELISA-like Immunoassay. Nanomaterials 2022, 12, 1630. [Google Scholar] [CrossRef] [PubMed]
- Pan, X.; Wang, H.; Li, C.; Zhang, J.Z.H.; Ji, C. MolGpka: A Web Server for Small Molecule pKa Prediction Using a Graph-Convolutional Neural Network. J. Chem. Inf. Model. 2021, 61, 3159–3165. [Google Scholar] [CrossRef]
- Wang, Z.; Zhang, R.; Yan, X.; Fan, K. Structure and Activity of Nanozymes: Inspirations for de Novo Design of Nanozymes. Mater. Today 2020, 41, 81–119. [Google Scholar] [CrossRef]
- Karpova, E.V.; Shcherbacheva, E.V.; Komkova, M.A.; Eliseev, A.A.; Karyakin, A.A. Core–Shell Nanozymes “Artificial Peroxidase”: Stability with Superior Catalytic Properties. J. Phys. Chem. Lett. 2021, 12, 5547–5551. [Google Scholar] [CrossRef] [PubMed]
- Zhang, Y.; Kudriashov, D.; Pershina, L.; Offenhäusser, A.; Mourzina, Y. Intrinsic Multienzyme-like Activities of the Nanoparticles of Mn and Fe Cyano-Bridged Assemblies. Nanomaterials 2022, 12, 2095. [Google Scholar] [CrossRef]
- Zhang, W.; Hu, S.; Yin, J.-J.; He, W.; Lu, W.; Ma, M.; Gu, N.; Zhang, Y. Prussian Blue Nanoparticles as Multienzyme Mimetics and Reactive Oxygen Species Scavengers. J. Am. Chem. Soc. 2016, 138, 5860–5865. [Google Scholar] [CrossRef] [PubMed]
- Huang, Y.; Xu, Q.; Zhang, J.; Yin, Y.; Pan, Y.; Zheng, Y.; Cai, X.; Xia, Q.; He, K. Prussian Blue Scavenger Ameliorates Hepatic Ischemia-Reperfusion Injury by Inhibiting Inflammation and Reducing Oxidative Stress. Front. Immunol. 2022, 13, 891351. [Google Scholar] [CrossRef]
- Komkova, M.A.; Ibragimova, O.A.; Karyakina, E.E.; Karyakin, A.A. Catalytic Pathway of Nanozyme “Artificial Peroxidase” with 100-Fold Greater Bimolecular Rate Constants Compared to Those of the Enzyme. J. Phys. Chem. Lett. 2021, 12, 171–176. [Google Scholar] [CrossRef]
- Bauduin, P.; Nohmie, F.; Touraud, D.; Neueder, R.; Kunz, W.; Ninham, B.W. Hofmeister Specific-Ion Effects on Enzyme Activity and Buffer pH: Horseradish Peroxidase in Citrate Buffer. J. Mol. Liq. 2006, 123, 14–19. [Google Scholar] [CrossRef]
- Lindmark, R.; Thorén-Tolling, K.; Sjöquist, J. Binding of Immunoglobulins to Protein A and Immunoglobulin Levels in Mammalian Sera. J. Immunol. Methods 1983, 62, 1–13. [Google Scholar] [CrossRef] [PubMed]
- Harlow, E.; Lane, D. Using Antibodies: A Laboratory Manual; CSHL Press: New York, NY, USA, 1999; ISBN 978-0-87969-544-6. [Google Scholar]
- Saha, D.; Acharya, D.; Dhar, T.K. Method for Homogeneous Spotting of Antibodies on Membranes: Application to the Sensitive Detection of Ochratoxin A. Anal. Bioanal. Chem. 2006, 385, 847–854. [Google Scholar] [CrossRef]
- Berg, S.; Kutra, D.; Kroeger, T.; Straehle, C.N.; Kausler, B.X.; Haubold, C.; Schiegg, M.; Ales, J.; Beier, T.; Rudy, M.; et al. Ilastik: Interactive Machine Learning for (Bio)Image Analysis. Nat. Methods 2019, 16, 1226–1232. [Google Scholar] [CrossRef]
- Stirling, D.R.; Swain-Bowden, M.J.; Lucas, A.M.; Carpenter, A.E.; Cimini, B.A.; Goodman, A. CellProfiler 4: Improvements in Speed, Utility and Usability. BMC Bioinform. 2021, 22, 433. [Google Scholar] [CrossRef]
- Ballesteros, C.A.S.; Mercante, L.A.; Alvarenga, A.D.; Facure, M.H.M.; Schneider, R.; Correa, D.S. Recent Trends in Nanozymes Design: From Materials and Structures to Environmental Applications. Mater. Chem. Front. 2021, 5, 7419–7451. [Google Scholar] [CrossRef]
- Lin, Y.; Zhao, A.; Tao, Y.; Ren, J.; Qu, X. Ionic Liquid as an Efficient Modulator on Artificial Enzyme System: Toward the Realization of High-Temperature Catalytic Reactions. J. Am. Chem. Soc. 2013, 135, 4207–4210. [Google Scholar] [CrossRef]
- Zhuang, J.; Midgley, A.C.; Wei, Y.; Liu, Q.; Kong, D.; Huang, X. Machine-Learning-Assisted Nanozyme Design: Lessons from Materials and Engineered Enzymes. Adv. Mater. 2023, 35, e2210848. [Google Scholar] [CrossRef]
Reagent | Volume, μL |
---|---|
Water | 150 |
Buffer | 100 |
2% H2O2 | 15 |
10 mg/mL DAB in H2O | 30 |
Nanozymes, 120 μg/mL | 5 |
Total volume | 300 |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2023 by the authors. 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/).
Share and Cite
Khramtsov, P.; Minin, A.; Galaeva, Z.; Mukhlynina, E.; Kropaneva, M.; Rayev, M. 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 2023, 28, 7622. https://doi.org/10.3390/molecules28227622
Khramtsov P, Minin A, Galaeva Z, Mukhlynina E, Kropaneva M, Rayev M. 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. 2023; 28(22):7622. https://doi.org/10.3390/molecules28227622
Chicago/Turabian StyleKhramtsov, Pavel, Artem Minin, Zarina Galaeva, Elena Mukhlynina, Maria Kropaneva, and Mikhail Rayev. 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, no. 22: 7622. https://doi.org/10.3390/molecules28227622