Identification and characterization of human monoclonal antibodies against the nuclear protein of severe fever with thrombocytopenia syndrome virus
Objective To screen human monoclonal antibodies(mAbs)against the nuclear protein(NP)of severe fever with thrombocytopenia syndrome virus(SFTSV),identify their binding specificity to both recombinant NP and NP in viral particles,and determine their affinity constant and binding kinetics.Methods Antibody genes were extracted from the blood of recovered individuals,and an antibody library was created using phage display.This library was panned by recombinant NP.The selected antibodies were expressed and purified.Enzyme linked immunosorbent assay(ELISA),western blot(WB),and indirect immunofluorescence assay(IFA)were used to assess the binding specificity of these mAbs to recombinant NP and NP in virions.Additionally,biolayer interferometry(BLI)was utilized to determine the antibody affinity constant.Results An antibody library with a capacity of 7.24×107 was successfully constructed.Following three rounds of panning,6 mAbs(named as NP-1,NP-10,NP-11,NP-20,NP-21,andNP-27)were isolated.The binding specificity of these 6 mAbs against recombinant NP was confirmed through indirect ELISA and WB analysis.Additionally,these mAbs were demonstrated specific-in binding to NP in virions as evidenced by IFA detection.The affinity constant values of the 6 mAbs,determined by BLI assay,ranged from 0.47 nmol/L to 32 nmol/L.Conclusions The 6 mAbs derived from the library are human mAbs that exhibit specificity to the NP of SFTSV and demonstrate a high affinity.These antibodies represent potential candidates for fundamental research and development of diagnostic reagents for SFTSV.
Severe fever with thrombocytopenia syndrome virusNuclear proteinPhage displayHuman monoclonal antibody
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