DEVELOPMENT AND PROPERTIES OF RECOMBINANT PROTEINS BASED ON THE BROADLY NEUTRALIZING ANTIBODY TO INFLUENZA A VIRUS

We studied the possibility of using a broadly neutralizing anti-influenza A antibody as a module for the development of different protein constructs for diagnostics. For this purpose we constructed two recombinant proteins — an antibody Fab-fragment and Fab-mCherry, which is a hybrid of the Fab-fragment and a fluorescent protein mCherry. Both proteins were expressed in Escherichia coli cells and purified in a functionally active state from cultivation medium. The antibody Fab-fragment was shown to bind all eleven tested strains of the influenza A H1N1 and H3N2 subtypes. The stronger binding was observed for the group I hemagglutinins that correlates with the immunochemical profile of the parental antibody. Comparison of the dissociation constants of complexes of the antibody Fab-fragment and Fab-mCherry with A(H1N1)/Solomon Islands/03/06 virus particles demonstrated that the attachment of mCherry protein did not interfere with the antigen-binding properties of the antibody Fab-fragment.

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