Skip to main content
SpringerLink
Log in

Catalytic nanocrystalline coordination polymers as an efficient peroxidase mimic for labeling and optical immunoassays

  • Original Paper
  • Published:
Microchimica Acta Aims and scope Submit manuscript

Abstract

We report that nanocrystalline Prussian blue of the type Fe4[Fe(CN)6]3 is a powerful peroxidase mimic for use in labeling of biomolecules. The cubic nanocrystals typically have a diameter of 15 nm and are capable of catalyzing the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine in the presence of H2O2 to form an intensively colored product with an absorption maximum at 662 nm. The determined pseudo turnover number is ~20,000 s−1 which is the highest value reported for nanoparticles of a size comparable to common proteins. We also present a method for the biotinylation of the surface of these nanocrystals, and show their use in competitive bioaffinity based assays of biotin and human serum albumin. The limits of detection are 0.35 and 0.27 μg mL−1, respectively. The results prove the applicability of coordination polymers for signal amplification and also their compatibility with the format of enzyme linked immunosorbent assays.

Nanocrystalline Prussian blue catalyzes the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine to form intensely colored products. Biotinylated cubic Prussian blue nanoparticles (biotin-NPs) facilitated the immunoassay of human serum albumin (HSA) utilizing biotinylated anti-human serum albumin antibody (biotin-IgG).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Wild D (2013) The immunoassay handbook. Theory and applications of ligand binding, ELISA and related techniques. Elsevier, Oxford, Fourth Edition

    Google Scholar 

  2. Kumari S, Dhar BB, Panda C, Meena A, Sen Gupta S (2014) Fe-TAML encapsulated inside mesoporous silica nanoparticles as peroxidase mimic: femtomolar protein detection. ACS Appl Mater Interfaces 6:13866–13873

    Article  CAS  Google Scholar 

  3. Jackson TM, Ekins RP (1986) Theoretical limitations on immunoassay sensitivity. Current practice and potential advantages of fluorescent Eu3+ chelates as Non-radioisotopic tracers. J Immunol Methods 87:13–20

    Article  CAS  Google Scholar 

  4. Walt DR (2013) Optical methods for single molecule detection and analysis. Anal Chem 85:1258–1263

    Article  CAS  Google Scholar 

  5. Lavery CB, MacInnis MC, MacDonald MJ, Williams JB, Spencer CA, Burke AA, Irwin DJG, DöCunha GB (2010) Purification of peroxidase from horseradish (Armoracia rusticana) roots. J Agric Food Chem 58:8471–8476

    Article  CAS  Google Scholar 

  6. Shokouhimehr M, Soehnlen ES, Khitrin A, Basu S, Huang SD (2010) Biocompatible Prussian blue nanoparticles: preparation, stability, cytotoxicity, and potential use as an MRI contrast agent. Inorg Chem Commun 13:58–61

    Article  CAS  Google Scholar 

  7. Gao L, Zhuang J, Nie L, Zhang J, Zhang Y, Gu N, Wang T, Feng J, Yang D, Perrett S, Yan X (2007) Intrinsic peroxidase-like activity of ferromagnetic nanoparticles. Nat Nanotechnol 2:577–583

    Article  CAS  Google Scholar 

  8. Wei H, Wang E (2013) Nanomaterials with enzyme-like characteristics (nanozymes): next-generation artificial enzymes. Chem Soc Rev 42:6060–6093

    Article  CAS  Google Scholar 

  9. Zhang Z, Zhu H, Wang X, Yang X (2011) Sensitive electrochemical sensor for hydrogen peroxide using Fe3O4 magnetic nanoparticles as a mimic for peroxidase. Microchim Acta 174:183–189

    Article  CAS  Google Scholar 

  10. Chang Q, Deng K, Zhu L, Jiang G, Yu C, Tang H (2009) Determination of hydrogen peroxide with the aid of peroxidase-like Fe 3O4 magnetic nanoparticles as the catalyst. Microchim Acta 165:299–305

    Article  CAS  Google Scholar 

  11. Chen W, Chen J, Liu A-L, Wang L-M, Li G-W, Lin X-H (2011) Peroxidase-like activity of cupric oxide nanoparticle. ChemCatChem 3:1151–1154

    Article  CAS  Google Scholar 

  12. Xu C, Qu X (2014) Cerium oxide nanoparticle: a remarkably versatile rare earth nanomaterial for biological applications. NPG Asia Mater 6 (3) art. no. e90

  13. Liu X, Wang Q, Zhao H, Zhang L, Su Y, Lv Y (2012) BSA-templated MnO2 nanoparticles as both peroxidase and oxidase mimics. Analyst 137:4552–4558

    Article  CAS  Google Scholar 

  14. Dong J, Song L, Yin J-J, He W, Wu Y, Gu N, Zhang Y (2014) Co3O4 nanoparticles with multi-enzyme activities and their application in immunohistochemical assay. ACS Appl Mater Interfaces 6:1959–1970

    Article  CAS  Google Scholar 

  15. He W, Liu Y, Yuan J, Yin J-J, Wu X, Hu X, Zhang K, Liu J, Chen C, Ji Y, Guo Y (2011) Au@Pt nanostructures as oxidase and peroxidase mimetics for use in immunoassays. Biomaterials 32:1139–1147

    Article  CAS  Google Scholar 

  16. Zhang Y, Lu F, Yan Z, Wu D, Ma H, Du B, Wei Q (2015) Electrochemiluminescence immunosensing strategy based on the use of Au@ Ag nanorods as a peroxidase mimic and NH4CoPO4 as a supercapacitive supporter. Microchim Acta 182:1421–1429

    Article  CAS  Google Scholar 

  17. Song Y, Qu K, Zhao C, Ren J, Qu X (2010) Graphene oxide: intrinsic peroxidase catalytic activity and its application to glucose detection. Adv Mater 22:2206–2210

    Article  CAS  Google Scholar 

  18. Qu R, Shen L, Chai Z, Jing C, Zhang Y, An Y, Shi L (2014) Hemin-block copolymer micelle as an artificial peroxidase and its applications in chromogenic detection and biocatalysis. ACS Appl Mater Interfaces 6:19207–19216

    Article  CAS  Google Scholar 

  19. Kitagawa S, Kitaura R, Noro S-I (2004) Functional porous coordination polymers. Angew Chem Int Ed 43:2334–2375

    Article  CAS  Google Scholar 

  20. Herren F, Fischer P, Ludi A, Hälg W (1980) Neutron diffraction study of Prussian blue, Fe4[Fe(CN)6]3·xH2O. Location of water molecules and long-range magnetic order. Inorg Chem 19:956–959

    Article  CAS  Google Scholar 

  21. Ricci F, Palleschi G (2005) Sensor and biosensor preparation, optimisation and applications of Prussian blue modified electrodes. Biosens Bioelectron 21:389–407

    Article  CAS  Google Scholar 

  22. Zhang W, Ma D, Du J (2014) Prussian Bue nanoparticles as peroxidase mimetics for sensitive colorimetric detection of hydrogen peroxide and glucose. Talanta 120:362–367

    Article  CAS  Google Scholar 

  23. Zhang X-Q, Gong S-W, Zhang Y, Yang T, Wang C-Y, Gu N (2010) Prussian blue modified iron oxide magnetic nanoparticles and their high peroxidase-like activity. J Mater Chem 20:5110–5116

    Article  CAS  Google Scholar 

  24. Hlavacek A, Bouchal P, Skládal P (2012) Biotinylation of quantum dots for application in fluoroimmunoassays with biotin-avidin amplification. Microchim Acta 176:287–293

    Article  CAS  Google Scholar 

  25. Josephy PD, Eling T, Mason RP (1982) The horseradish peroxidase-catalyzed oxidation of 3,5,3′,5′-tetramethylbenzidine. Free radical and charge-transfer complex intermediates. J Biol Chem 257:3669–3675

    CAS  Google Scholar 

  26. Hlaváček A, Sedlmeier A, Skládal P, Gorris HH (2014) Electrophoretic characterization and purification of silica-coated photon-upconverting nanoparticles and their bioconjugates. ACS Appl Mater Interfaces 6:6930–6935

    Article  Google Scholar 

  27. Shang J, Gao X (2014) Nanoparticle counting: towards accurate determination of the molar concentration. Chem Soc Rev 43:7267–7278

    Article  CAS  Google Scholar 

  28. Metelitza DI, Karasyova EI (2002) Activation of peroxidase-catalyzed oxidation of 3,3′,5,5′-tetramethylbenzidine with poly(salicylic acid 5-aminodisulfide). Biochem Mosc 67:1048–1054

    Article  CAS  Google Scholar 

  29. Xia Y, Nguyen TD, Yang M, Lee B, Santos A, Podsiadlo P, Tang Z, Glotzer SC, Kotov NA (2011) Self-assembly of self-limiting monodisperse supraparticles from polydisperse nanoparticles. Nat Nanotechnol 6:580–587

    Article  CAS  Google Scholar 

  30. Kendall C, Ionescu Matiu I, Dreesman GR (1983) Utilization of the biotin/avidin system to amplify the sensitivity of the enzyme-linked immunosorbent assay (ELISA). J Immunol Methods 56:329–339

    Article  CAS  Google Scholar 

  31. Zhao L, Lin J-M, Li Z (2005) Comparison and development of two different solid phase chemiluminescence ELISA for the determination of albumin in urine. Anal Chim Acta 541:199–207

    Article  CAS  Google Scholar 

  32. Kessler MA, Meinitzer A, Peter W, Wolfbeis OS (1997) Microalbuminuria and borderline-increased albumin excretion determined with a centrifugal analyzer and the albumin blue 580 fluorescence assay. Clin Chem 43:996–1002

    CAS  Google Scholar 

  33. Doumas BT, Peters Jr T (1997) Serum and urine albumin: a progress report on their measurement and clinical significance. Clin Chim Acta 258:3–20

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors acknowledge funding by the Program of “Employment of Newly Graduated Doctors of Science for Scientific Excellence” (Grant CZ.1.07/2.3.00/30.0009), cofinanced by the European Social Fund and the state budget of the Czech Republic. The work was also supported by the CEITEC - Central European Institute of Technology (CZ.1.05/1.1.00/02.0068) from the European Regional Development Fund, COST CZ LD15023 from “The Ministry of Education, Youth and Sports” of the Czech Republic and by funds from the Faculty of Medicine of the Masaryk University (MUNI/A/1558/2014). Access to the core facilities of CEITEC CryoEM is acknowledged.

Author contributions

The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript. Veronika Čunderlová and Antonín Hlaváček contributed equally.

Author information

Authors and Affiliations

  1. CEITEC - Central European Institute of Technology, Group Nanobiotechnology, Masaryk University, Kamenice 5/A35, 625 00, Brno, Czech Republic

    Veronika Čunderlová, Antonín Hlaváček, Veronika Horňáková, Miroslav Peterek, Daniel Němeček & Petr Skládal

  2. Faculty of Medicine, Department of Histology and Embryology, Masaryk University, 625 00, Brno, Czech Republic

    Aleš Hampl

  3. Department of Virology, Veterinary Research Institute, 62100, Brno, Czech Republic

    Luděk Eyer

Authors
  1. Veronika Čunderlová
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Antonín Hlaváček
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Veronika Horňáková
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Miroslav Peterek
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Daniel Němeček
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Aleš Hampl
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Luděk Eyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Petr Skládal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Antonín Hlaváček.

Electronic supplementary material

ESM 1

(PDF 816 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Čunderlová, V., Hlaváček, A., Horňáková, V. et al. Catalytic nanocrystalline coordination polymers as an efficient peroxidase mimic for labeling and optical immunoassays. Microchim Acta 183, 651–658 (2016). https://doi.org/10.1007/s00604-015-1697-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00604-015-1697-z

Keywords

Search

Navigation