应用噬菌体表面展示技术筛选流感病毒H3N2抗原模拟表位

Epitope screening of influenza A （H3N2） by using phage display library

目的筛选特异性流感病毒H3N2抗原模拟表位，为开展新的流感病毒疫苗研究探索新的途径。方法应用噬菌体表面展示技术，以抗．H3N2的单克隆抗体作为固相筛选分子，对人工合成的噬菌体随机肽库进行5轮“吸附-洗脱-扩增”的筛选过程，在第5轮筛选后，随机挑取48个克隆，经噬菌体酶联免疫吸附法（ELISA）鉴定并进行交叉反应实验以及竞争抑制性实验，最后对所选克隆进行DNA序列分析，以确定H3N2抗原的模拟表位。结果经噬菌体富集后，从随机筛选的48个克隆中得到21个阳性克隆，经ELISA鉴定及交叉反应实验、竞争抑制性实验后，确定氨基酸序列XTXPYXX为H3N2的模拟表位。结论用噬菌体7肽库筛选得到H3N2的模拟表位，为开展用流感病毒模拟表位探索新的防治方法研究奠定了基础。

Objective To screen the influenza A （H3N2） mimotopes by using phage display library. Methods Using influenza A （H3N2） monoclonal antibody as selective molecule, a 7 mer phage peptide library was biopanned and positive clones were selected by ELISA, competition assay and DNA sequencing. Results 21 positive clones were chosen for DNA sequencing. From the experiment and sequencing comparison results, one epitope was comfirmed as mimotope of influenza A （H3N2）. Conclusion Influenza A （H3N2） mimotope was obtained by phage peptide library screening. The result provide a new approach for new Influenza virals vaccine development.