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

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Abstract

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.

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Acknowledgements

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

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Authors and Affiliations

  1. Ural Federal University, Chemical Technology Institute, Analytical Chemistry Department, 19 Mira str., Ekaterinburg, 620002, Russia

    Tatiana S. Svalova, Margarita V. Medvedeva, Anna A. Saigushkina, Natalya N. Malysheva, Valeria O. Zhdanovskikh, A. V. Okhokhonin & Alisa N. Kozitsina

  2. V.A. Trapeznikov Institute of Control Sciences RAS, Ulitsa Profsoyuznaya, 65, Moscow, 117997, Russia

    Ivan V. Kozitsin

  3. Moscow Institute of Physics and Technology, Institutskiy Pereulok 9, Dolgoprudny, 141701, Russia

    Ivan V. Kozitsin

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  1. Tatiana S. Svalova
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  2. Margarita V. Medvedeva
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  3. Anna A. Saigushkina
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  4. Ivan V. Kozitsin
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  5. Natalya N. Malysheva
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  7. A. V. Okhokhonin
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Correspondence to Tatiana S. Svalova.

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Published in the topical collection Euroanalysis XX with guest editor Sibel A. Ozkan.

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Svalova, T.S., Medvedeva, M.V., Saigushkina, A.A. et al. A label-free impedimetric immunosensor based on covalent immobilization of anti-E. Coli antibody via a copper-catalyzed azide-alkyne cycloaddition reaction. Anal Bioanal Chem 412, 5077–5087 (2020). https://doi.org/10.1007/s00216-019-02381-1

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