Development of virus-specific anti-NS1 antibodies and expression system for flavivirus recombinant NS1 for use in neurotropic flavirus diagnostic, therapeutics, and vaccination development

West Nile virus (WNV) and Usutu virus (USUV) are two mosquito-borne flaviviruses that are emerging in Europe, posing an increasing health risk. Given the increasingly imminent danger that these viruses represent, it is necessary to find both more efficient methods for early diagnosis and therapeutic or preventive solutions to curb the impact of these pathogens on human health. Non-structural protein 1 (NS1) of WNV and USUV is considered a promising target for diagnostic, vaccine, and therapeutic development. The project aims to produce monoclonal antibodies (mAbs) that are highly specific for NS1 of WNV and USUV using a subtractive immunization approach and to set up an expression system for recombinant flavivirus proteins that preserve the original oligomerization state of NS1, which circulates in the blood as a hexamer. The generated mAbs will drive the development of the next generation of diagnostics, and this will give us a competitive edge toward follow-up project applications and negotiations with diagnostic assay developers. Furthermore, the antibodies and recombinant proteins can contribute to elucidating the role of NS1 in vaccine and therapeutics development, constituting the necessary preliminary data to proceed with broader project proposals.