Electrochemical immunosensors based on gold nanoparticles for the determination of ovalbumin in immunobiological preparations

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Аннотация

A sandwich-type voltammetric immunosensor based on conjugates of gold nanoparticles (AuNP) with specific immunoglobulins (IgG) against ovalbumin (IgG@AuNP) is developed for the determination of ovalbumin in some immunobiological preparations. The IgG@AuNP conjugate is synthesized by passive adsorption. A carbon-containing planar printed electrode modified by laser reduced graphene oxide is used as a substrate electrode for immobilizing the receptor layer of specific immunoglobulins. A possibility of the catalytic reduction of silver nitrate with a mixture of reducing agents, citric acid and metol, is considered in order to enhance the signal of AuNP. Conditions for the voltammetric recording of the electrochemical signal of silver on an immunosensor, which has been successfully tested in determining ovalbumin residues in some immunobiological drugs, are selected. Enzyme-linked immunosorbent assay is used as a reference method.

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Авторлар туралы

Е. Dorozhko

National Research Tomsk Polytechnic University

Хат алмасуға жауапты Автор.
Email: evd@tpu.ru
Ресей, 634050, Tomsk, Lenin Avenue 30

A. Solomonenko

National Research Tomsk Polytechnic University

Email: evd@tpu.ru
Ресей, 634050, Tomsk, Lenin Avenue 30

M. Saqib

National Research Tomsk Polytechnic University

Email: evd@tpu.ru
Ресей, 634050, Tomsk, Lenin Avenue 30

V. Semin

Institute of Strength Physics and Materials Science, Siberian Branch, Russian Academy of Sciences

Email: evd@tpu.ru
Ресей, 634055, Tomsk, Academic Avenue, 2/4

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2. 1. Diagram of an electrochemical immunosensor for the determination of ovalbumin in vaccines.

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3. Fig. 2. Light absorption spectra in the visible and UV regions in bidistilled water (1) of Au (2) NPS and IgG@Au (3)NPS conjugates.

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4. Fig. 3. Transmission electron microscopy images of (a) Au NPS and (b) IgG conjugates@Au NPS (IgG contrast with 2.0% uranyl acetate solution).

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5. 4. Cyclic voltammograms (a) HCAi of IgG@HCAi conjugates on a carbon–containing planar printed electrode in 0.1 M HCl-NaCl background electrolyte solution (1 :1, by volume), v = 100 mV/s, before immobilization of the receptor layer (1), after immobilization of the receptor layer (2); (b) cyclic voltammograms of silver after manifestation on the surface of an electrochemical immunosensor in different background electrolytes: 1 – 0.05 M HCl–0.05 M NaClO4 solution (1 : 1), 2 – 0.25 M KNO3 solution–0.5 M HNO3 (1 : 1), the volume of the background electrolyte is 200 µl.

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6. Fig. 5. (a) Image of the surface of the immunosensor obtained by scanning electron microscopy after catalytic reduction of silver with a mixture of 0.2% citric acid and 0.25% methol on HCAi conjugates IgG@HCAi (electron backscattering mode of the surface); (b) the EMF spectrum of the immunosensor surface after catalytic reduction of silver with characteristic X-ray lines of C, O, Au and Ag.

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7. 6. Calibration dependence for the determination of ovalbumin using an electrochemical immunosensor in the concentration range from 0.125 to 128 ng/ml (insert – anodic inversion voltammograms of silver oxidation).

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