Abstract
BackgroundIn this work, we report an electrochemical biosensor for the detection of anti-hemagglutinin antibodies against the swine virus H1N1 present in mice sera immunized with mixture of His6-H1 HA in monomeric and oligomeric form. The oriented immobilization of the recombinant His-tagged hemagglutinin (His6-H1 HA) consists of: (i) formation of a mixed layer of 4-mercaptobutanol (MBT) and the thiol derivative of dipyrromethene (DPM); (ii) complexation of Cu (II) by DPM; (iii) immobilization of His6-H1 HA via coordination bonds between Cu (II) sites from DPM–Cu (II) complex and imidazole nitrogen atoms of a histidine tag; (iv) filling free spaces with bovine serum albumin. The interactions between recombinant His6- H1 HA covalently attached to the electrode surface and the anti-hemagglutinin H1 antibodies present in mice sera were explored with Osteryoung square-wave voltammetry.ResultsThis analytical device was able to detect the antibodies present in vaccinated mice sera diluted from 1 × 109 to 1 × 108 fold.ConclusionsThe unprecedented sensitivity of described biosensor is much better than widely use ELISA test and other analytical methods for determination of antibodies against the influenza A viruses. It has been proved that redox active DPM-Cu (II) monolayer is a universal platform suitable for stable and oriented immobilization of any His-tagged sensing elements. Thus, this universal layer could be a base of numerous analytical devices suitable for detection of antibodies against different viruses.
Highlights
In this work, we report an electrochemical biosensor for the detection of anti-hemagglutinin antibodies against the swine virus H1N1 present in mice sera immunized with mixture of His6-H1 HA in monomeric and oligomeric form
In our previously published paper [36], we have proposed the redox active layer consisted with dipyrromethene (DPM)–Cu (II) complex for stable and oriented immobilization of histidine – tagged fragment of avian influenza H5N1 virus
This strategy has been already applied for biosensor assembly destined for detection of antibodies against H5N1 viruses present in vaccinated hen sera [36]
Summary
We report an electrochemical biosensor for the detection of anti-hemagglutinin antibodies against the swine virus H1N1 present in mice sera immunized with mixture of His6-H1 HA in monomeric and oligomeric form. Influenza viruses induce annual epidemics and casual pandemics that have claimed the lives of millions. Their very high seasonal variation makes effective vaccination and environment control very challenging [1,2,3,4,5]. Transmission of the virus from pigs to humans does not always cause flu, sometimes results only in the creation of antibodies in the blood [17] Record of these outbreaks and real-time monitoring of the evolution of this virus are important for the infectious disease control programs and for better understanding of the factors that specify viral pathogenicity and transmissibility
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