从噬菌体随机肽库中筛选流感病毒神经氨酸酶的抑制多肽

目的:从噬菌体呈现12肽库中筛选与流感病毒神经氨酸酶特异性结合的肽。方法:以甲三型流感病毒裂解疫苗原液为靶分子,经过3轮生物淘选,从噬菌体随机肽库中筛选与之结合的噬菌体。用ELISA方法鉴定噬菌体克隆与靶分子的结合力,用荧光方法测定噬菌体克隆对流感病毒A/Sydney/5/97(H3N2)神经氨酸酶的抑制活性。对筛选到的阳性克隆进行DNA序列测定并推导出相应的氨基酸序列。结果:经过3轮筛选后,42个噬菌体克隆与靶分子有高度亲和力,23个噬菌体克隆对流感病毒A/Sydney/5/97(H3N2)神经氨酸酶有抑制活性。对27个噬菌体克隆的测序结果表明,分别有10个和2个克隆的序列是一致的,其氨基酸序列分别为KSLSRHDHIHHH和WPRHHHSASVQT。结论:通过噬菌体肽库筛选到抑制流感病毒神经氨酸酶的12肽,为进一步研究对流感病毒神经氨酸酶有抑制活性的分子药物奠定了基础。

Peptide Inhibitors of Influenza Virus Neuraminidase Selected from Phage Random Peptide Library

Objective: To select the peptide binding to influenza virus neuraminidase from phage display 12 peptide library. Methods: Using the influenza A3 split vaccine as the target molecule, peptides that could bind with target molecule were selected from the phage display 12 peptide library by 3 rounds of biopaning. The affinity of selected peptides were assayed by ELISA and the inhibitory activity to influenza virus A/Sydney/5/97 neuraminidase were detected by fluorometric method. Then, the positive clones were used for sequencing, and the amino acid sequences of polypeptide displayed on phage were deduced. Results: 42 positive clones with high affinity were obtained and 23 positive clones could inhibit the influenza virus A/Sydney/5/97 neuraminidase activity after 3 rounds of biopaning. Sequencing of the gene encoding 27 positive peptides showed that there were 10 and 2 identical sequences, and the amino acid sequences were KSLSRHDHIHHH, WPRHHHSASVQT respectively. Conclusion: Potential 12 peptides against influenza virus neuraminidase could be selected from phage display peptide library, which provide the basis for novel anti-neuraminidase agents.