Epitope Mapping of HTNV NP Using mAb 5H5 from a Phage-displayedRalldom Nonapeptide Ia’brary

To screen the antigenic epitope with monoclonal antibody (mAb)5H5 against Hantaan virus nucleocapsid protein from phage-displayed constrained nonapeptide library,we have completed the work as follows.(1) Purification and biotinylation of mAb 5H5.(2)Biopanning of the phage peptide library by using the mAb 5H5 as selective moleculars.(3)Identification of the epitope.The positive clones were confirmed by sandwich ELISA,competition ELISA and DNA sequencing.Amino acid sequences of 10 positive clones were compared with the nucleocapsid protein of Hantaan virus strain 76 118.The ten positive clones were selected by mAb 5H5 and the binding between the short peptide and the mAb can be inhabited by natural Hantaan virus antigen.Amino acid sequences of 10 positive clones were consensus:VRDAEEQYE,that is roughly identical with the sequence of amino acid of N terminal of the nucleocapsid protein.These results demonstrate that the linear epitope is the antigenic epitope of Hantaan virus nucleocapsid protein and the rando