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目的 应用噬菌体展示肽文库筛选可与汉坦病毒囊膜蛋白中和性单抗 (mAb)F3特异性结合的配体肽。方法以F3mAb为筛选配基 ,对噬菌体展示的随机 12肽文库进行 3轮生物亲和淘选 ;用夹心ELISA和竞争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轮生物淘选 ,能被抗体捕获的噬菌体克隆为 2 1/ 2 2。ELISA测定显示 ,筛选到的噬菌体短肽能与F3mAb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/SEOVM蛋白G2区第 75 0～ 75 9位氨基酸有较高的同源性。结论 本研究为基于表位水平的HFRSV特异性分子多肽疫苗的设计提供了重要的依据分析。结果 通过 3轮生物淘选 ,能被抗体捕获的噬菌体克隆为 2 1/ 2 2。ELISA测定显示 ,筛选到的噬菌体短肽能与F3mAb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/SEOVM蛋白G2区第 75 0～ 75 9位氨基酸有较高的同源性。结论 本研究为基于表位水平的HFRSV特异性分子多肽疫苗的设计提供了重要的依据
OBJECTIVE: To screen phage display peptide libraries for ligand peptides that specifically bind to the Hantavirus envelope protein neutralizing monoclonal antibody (mAb) F3. Methods F3mAb was used as the screening ligand to perform 3 rounds of biophilic panning on phage displayed random 12-peptide library. The binding characteristics of the clones were screened by sandwich ELISA and competitive ELISA, and the positive clones were further sequenced and analyzed. Results Three rounds of biopanning showed that the number of phage clones that could be captured by the antibody was 2 1/2 2. ELISA assay showed that the screened phage short peptides could specifically bind F3mAb. Sequence analysis showed that the seven positive clones had the same amino acid sequence and all of them were MHGPTKNQMWHT. Homology analysis showed that this sequence shared high homology with amino acids 75 0 ~ 75 9 in G2 region of HTNV / SEOVM protein. Conclusion This study provides an important basis for the design of HFRSV-based peptide vaccine based on epitope level. Results Three rounds of biopanning showed that the number of phage clones that could be captured by the antibody was 2 1/2 2. ELISA assay showed that the screened phage short peptides could specifically bind F3mAb. Sequence analysis showed that the seven positive clones had the same amino acid sequence and all of them were MHGPTKNQMWHT. Homology analysis showed that this sequence shared high homology with amino acids 75 0 ~ 75 9 in G2 region of HTNV / SEOVM protein. Conclusion This study provides an important basis for the design of HFRSV-specific peptide vaccine based on epitope level