A voltammetric sandwich immunosensor based on gold nanoparticles (AuNPs) conjugates and specific immunoglobulins (IgG@AuNPs) has been developed for the quantitative determination of inactivated hepatitis A virus (HAV) in a vaccine. For the first time, copper nanoparticles (CuNPs) were catalytically reduced from copper sulfate on the surface of AuNPs conjugates by a chemical reducing agent, reduced nicotinamide adenine dinucleotide (NADH). Control experiments showed that copper was not formed in the absence of AuNPs on the electrode surface or in the absence of NADH. The AuNPs and IgG@AuNPs conjugates before and after the reduction of CuNPs were characterized by transmission electron microscopy (TEM). A carbon-based screen-printed electrode (SPE), functionalized with laser reduced graphene oxide (LRGO) to enhance the conductivity of its working surface, was employed as the solid substrate for immobilizing the receptor layer. This layer consisted of 'capture' antibodies derived from rabbits immunized with HAV. During the development of the electrochemical immunosensor, particular attention was paid to the optimization of the conditions for the deposition time of copper on the surface of the gold particles of the IgG@AuNPs conjugates and the concentration of the reducing agent NADH. The elemental composition and electronic state of the atoms on the surface of the electrodes modified with AuNPs before and after the reduction of CuNPs with NADH were investigated by X-ray photoelectron spectroscopy (XPS). This study demonstrates that the electrochemical immunosensor amplifies the analytical signal from copper electrooxidation in biosensor for detecting HAV in vaccines. Concurrently, it reduces immunoassay costs and viral antigen detection time relative to traditional enzyme-linked immunosorbent assay (ELISA) systems.

Division for Chemical Engineering School of Earth Sciences and Engineering National Research Tomsk Polytechnic University Lenin Avenue 30 634050 Tomsk Russia

Division for Chemical Engineering School of Earth Sciences and Engineering National Research Tomsk Polytechnic University Lenin Avenue 30 634050 Tomsk Russia; UNESCO Laboratory of Environmental Electrochemistry Department of Analytical Chemistry Faculty of Science Charles University Albertov 6 CZ 12843 Prague 2 Czech Republic

Moscow Institute of Physics and Technology 141701 Dolgoprudny Russia

Research School of Chemistry and Applied Biomedical Sciences National Research Tomsk Polytechnic University Lenin Avenue 30 634050 Tomsk Russia

State Research Center of Virology and Biotechnology Vector Federal Service for Surveillance on Consumer Rights Protection and Human Well being Koltsovo 630559 Novosibirsk Russia

АО Vector BiAlgam Koltsovo 630559 Novosibirsk Russia

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