A label-free impedimetric immunosensor based on covalent immobilization of anti-E. Coli antibody via a copper-catalyzed azide-alkyne cycloaddition reaction

  • Tatiana S. SvalovaEmail author
  • Margarita V. Medvedeva
  • Anna A. Saigushkina
  • Ivan V. Kozitsin
  • Natalya N. Malysheva
  • Valeria O. Zhdanovskikh
  • A. V. Okhokhonin
  • Alisa N. Kozitsina
Research Paper
Part of the following topical collections:
  1. Euroanalysis XX


In this article, an original method is proposed for address and covalent immobilization of anti-E. coli antibodies on a screen-printed electrode (SPE). The method is based on a copper-catalyzed “click” reaction between a polyvinylbenzylazide (PVBA) film electrochemically deposited on the electrode surface and acetylene fragments of propargyl-N-hydroxysuccinimide ester. The products of electrochemical oxidation of copper particles incorporated in the polymer film on the electrode were first used for catalysis of the click reaction. This approach allowed us to reduce the immobilization time from a few hours for conventional methods to just 30 min, and to prevent denaturation of the immunoreceptor. The modified electrodes were characterized by scanning electron microscopy (SEM), electrochemical impedance spectroscopy (EIS), and cyclic voltammetry (CV). Based on the results obtained, a label-free impedimetric immunosensor for E. coli detection was developed. The detection limit of the immunosensor was estimated as 6.3 CFU/ml, with a linear range of 103–106 CFU/ml. The immunosensor demonstrated good stability during 30 days of storage in phosphate buffer solution (PBS, pH 7) and selectivity toward excess Staphylococcus aureus bacteria.


Antibody covalent immobilization Copper-catalyzed azide-alkyne cycloaddition Label-free impedimetric immunosensor E. coli 



The research was supported by the Russian Science Foundation (grant number 18-73-00224).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

216_2019_2381_MOESM1_ESM.pdf (247 kb)
ESM 1 (PDF 247 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2020

Authors and Affiliations

  • Tatiana S. Svalova
    • 1
    Email author
  • Margarita V. Medvedeva
    • 1
  • Anna A. Saigushkina
    • 1
  • Ivan V. Kozitsin
    • 2
    • 3
  • Natalya N. Malysheva
    • 1
  • Valeria O. Zhdanovskikh
    • 1
  • A. V. Okhokhonin
    • 1
  • Alisa N. Kozitsina
    • 1
  1. 1.Ural Federal University, Chemical Technology Institute, Analytical Chemistry DepartmentEkaterinburgRussia
  2. 2.V.A. Trapeznikov Institute of Control Sciences RASMoscowRussia
  3. 3.Moscow Institute of Physics and TechnologyDolgoprudnyRussia

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