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为研究日本脑炎病毒 (JEV )E蛋白模拟肽 ,将抗JEVE蛋白的mAb 2H4淘筛噬菌体 15肽库。经夹心ELISA、竞争ELISA鉴定后 ,随机挑取 10个阳性克隆 ,测序并与JEVE蛋白同源比较。将阳性噬菌体免疫小鼠 ,检测血清中特异性抗体。ELISA结果显示筛选到的噬菌体能特异地与mAb 2H4结合 ,并且这种结合可被JEV天然抗原所竞争抑制。 10个阳性克隆的氨基酸序列相同 :—RQDPQWPYANSTIAR— ,同源分析得到的序列STXAR可能为mAb 2H4识别的模拟表位。阳性噬菌体表达的 15肽能够刺激小鼠产生特异性抗体。该噬菌体表达短肽模拟JEVE蛋白的部分抗原性。
To study Japanese encephalitis virus (JEV) E-protein mimetic peptide, anti-JEVE protein mAb 2H4 was screened for a phage 15 peptide library. After sandwich ELISA and competitive ELISA, 10 positive clones were randomly selected, sequenced and compared with JEVE proteins. The positive phages were immunized to detect serum specific antibodies. ELISA results showed that the selected phage could specifically bind to mAb 2H4, and this binding could be competitively inhibited by JEV native antigen. The 10 positive clones had the same amino acid sequence: -RQDPQWPYANSTIAR-, and the sequence STXAR obtained by homology analysis may be a mimotope recognized by mAb 2H4. Positive phage-expressed peptide 15 can stimulate mice to produce specific antibodies. This phage expression short peptide mimics the partial antigenicity of the JEVE protein.