Unusual switchable peroxidase-mimicking nanozyme for the determination of proteolytic biomarker

  • Claire McVey
  • Natasha Logan
  • Nguyen T. K. Thanh
  • Christopher Elliott
  • Cuong Cao
Open Access
Research Article


Detection of enzyme biomarkers originating from either bio-fluids or contaminating microorganisms is of utmost importance in clinical diagnostics and food safety. Herein, we present a simple, low-cost and easy-to-use sensing approach based on the switchable peroxidase-mimicking activity of plasmonic gold nanoparticles (AuNPs) that can catalyse for the oxidation of 3,3’,5’5-tetramethylbenzidine (TMB) for the determination of protease enzyme. The AuNP surface is modified with casein, showing dual functionalities. The first function of the coating molecule is to suppress the intrinsic peroxidase-mimicking activity of AuNPs by up to 77.1%, due to surface shielding effects. Secondly, casein also functions as recognition sites for the enzyme biomarker. In the presence of protease, the enzyme binds to and catalyses the degradation of the coating layer on the AuNP surface, resulting in the recovery of peroxidase-mimicking activity. This is shown visually in the development of a blue colored product (oxidised TMB) or spectroscopically as an increase in absorbance at 370 and 650 nm. This mechanism allows for the detection of protease at 44 ng·mL−1 in 90 min. The nanosensor circumvents issues associated with current methods of detection in terms of ease of use, compatibility with point-of-care testing, low-cost production and short analysis time. The sensing approach has also been applied for the detection of protease spiked in ultra-heat treated (UHT) milk and synthetic human urine samples at a limit of detection of 490 and 176 ng·mL−1, respectively, showing great potential in clinical diagnostics, food safety and quality control.


gold nanoparticles peroxidase-mimicking nanozyme enzyme detection biomarkers 



The author C. M. and N. L. thank the PhD studentship support from the Department of Employment and Learning for Northern Ireland (DEL); C. C. thanks the strong support from the Central Research Support Funds of Queen’s University Belfast via a start-up grant, the support from the Agri-Food Quest Competence Centre R&D funding programme sponsored by Invest Northern Ireland Agency (Invest NI), and the support from the Queen’s University of Belfast AMR Network (QUBAN) sponsored by the UK’s Engineering and Physical Sciences Research Council (EPSRC).

Supplementary material

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Unusual switchable peroxidase-mimicking nanozyme for the determination of proteolytic biomarker


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© The authors 2018

Open Access: This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Authors and Affiliations

  • Claire McVey
    • 1
  • Natasha Logan
    • 1
  • Nguyen T. K. Thanh
    • 2
    • 3
  • Christopher Elliott
    • 1
  • Cuong Cao
    • 1
    • 4
  1. 1.Institute for Global Food Security, School of Biological SciencesQueen’s University BelfastBelfastUK
  2. 2.Biophysics Group, Department of Physics and AstronomyUniversity College LondonLondonUK
  3. 3.UCL Healthcare Biomagnetic and Nanomaterials LaboratoriesLondonUK
  4. 4.Material and Advanced Technologies for HealthcareQueen’s University BelfastBelfastUK

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