应用噬菌体展示随机肽库淘筛mAb 5H5识别的抗原表位

本文利用噬菌体随机9肽库探索汉滩病毒（HTNV）核衣壳蛋白（NP）B细胞抗原表位。以抗HTNV NP单克隆抗体(mAb）5H5作为筛选分子，生物淘洗噬菌体递呈的随机9肽库。阳性克隆经夹心ELISA、竞争ELISA鉴定后，随机挑取10个克隆，DNA测序，与HTNV76－118株S基因进行同源性分析。结果显示筛选到的噬菌体能特异地与5H5结合，这种结合可被天然抗原所抑制。10个克隆的氨基酸序列相同，均为VRDAEEQYE，与76－118株NP氨基端的aa25-33一致。证实了该线性表位是mAb 5H5识别的表位，噬菌体肽库有助于病毒抗原表位的确定。

Epitope Mapping of HTNV NP Using mAb 5H5 from a Phage-displayed Random Nonapeptide Library

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 random peptides library is helpful to identification of epitopes.