应用噬菌体肽文库筛选McAb F3特异性结合肽

目的：应用噬菌体展示肽文库筛选可与抗汉坦病毒囊膜蛋白单抗F3特异性结合的配体肽。方法：采用protein-A亲和层析纯化的F3单抗为筛选配基，对噬菌体展示的随机12肽文库进行生物亲和淘洗，夹心ELISA、竞争ELISA鉴定筛选克隆的结合特性，并进一步对阳性克隆进行序列测定和分析。结果：通过3轮生物淘洗，能被抗体捕获的噬菌体克隆为91．7％（11／12）；ELISA测定显示筛选到的噬菌体短肽能与F3单抗特异性结合；序列分析表明7个阳性克隆氨基酸序列相同，均为－MHGPTKNQMWHT，同源性分析显示该序列与HTNV／SEOV M蛋白G2区第750－759位氨基酸有较高的同源性，结论：获得了具有良好结合活性的模拟表位肽，为基于表位水平的HFRSV（肾病综合征出血热病毒）特异性分子多肽疫苗的设计提供了重要依据。

Study on peptide mimicking epitope of Hantaan virus glycoprotein by phage display peptide library

Objective To obtain the 12-mer phage clones displaying the Hantaan virus mimic epitope related to neutralizing antibody. Methods A group-specific McAb F3 against glycoprotein of both HTNV and SEOV, which possesses fairly high neutralizing titer and has strong animal protective activity. Using F3 McAb as selective molecule, a 12-mer phage peptide library was biopanned for 3 rounds, and the result was confirmed by sandwich ELISA, competition ELISA and DNA sequencing. Results After 3 rounds of effective screening, 11 positive clones screened out from 12 clones were selected, and amino acid sequences of 7 positive clones were consensus: MHGPTKNQMWHT, which was similar to G2 750TKYQYPWHT 759. Conclusion The peptides displayed by phage can mimic the epitope of HFRSV antigen, which provide the potentiality for preparing more effective epitope-based vaccine. [