从噬菌体15肽库中筛选乙脑病毒糖蛋白模拟肽

目的 用抗JEVE蛋白的mAb2H4筛选噬菌体15肽库，以研究JEVE蛋白模拟肽。方法 用生物素标记的mAb2H4，对噬菌体随机15肽库进行3轮筛选，经ELISA鉴定后，随机挑战取10个阳性噬菌体克隆测序，并与JEVE蛋白进行同源性比较，结果 筛选到的噬菌体能与mAb2H4特异地结合，并且这种结合可被天然JEV抗原所抑制，10个阳性噬菌体克隆的氨基酸序列相同，均为RQD-PQWPYANSTIAR，同源分析表明，在JEVE蛋白不同区域有两个同源性较高的序列：STXAR和WXXAXST，阳性噬菌体展示肽了能与抗天然JEV抗原的鼠血清产生特异性反应。结论 筛选的该噬菌体克隆展示肽可模拟JEVE蛋白的部分抗原性。

Screening of mimicry peptide of Japanese encephalitis virus E protein from phage 15-mer peptide library

Aim To study the mimicry peptide of JEV E protein by screening a phage 15-mer peptide library with anti-JEV E protein mAb 2H4. Methods After three rounds biopanning, the enriched positive phage clones were identified by ELISA. 10 positive phage clones were sequenced and compared homologously with JEV E protein. Results The short peptide displayed on screened positive phage could bind specifically to mAb 2H4, and the binding could be inhibited by natural JEV Ag. Amino acid sequences of the 10 positive phage clones were consensus, that is, RQDPQWPYANSTIAR. By homology analysis, two higher homologous sequences STXAR and WXXAXST were found in different regions of JEV E protein. The peptide displayed on positive phage could react specifically with the mouse antiserum against natural JEV Ag . Conclusion This peptide displayed on positive phage may mimic partial antigenicity of JEV E protein.