Phage-displayed peptides mimicking the discontinuous neutralization sites of Puumala hantavirus envelope glycoproteins
Phage-displayed peptides mimicking the discontinuous neutralization sites of Puumala hantavirus envelope glycoproteins
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DOI：
10.1006/viro.1999.9930
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发表时间：
1999-09-30
期刊：
影响因子：
3.7
通讯作者：
Lankinen, H
中科院分区：
文献类型：
--
作者：
Heiskanen, T;Lundkvist, Å;Lankinen, H
我们选择肽配体模仿的表面结构的不连续的结合位点的普马拉汉坦病毒中和单克隆抗体从随机的18个氨基酸的肽库中含有一个二硫键在一个固定的位置，并显示在丝状噬菌体。通过缩短结合时间或通过与抗原竞争洗脱来改变选择条件，对于选择可以与包膜糖蛋白G1和G2的一级序列比对的肽插入物是至关重要的。相应地，当将包膜糖蛋白序列合成为膜上斑点的重叠肽时，发现与噬菌体展示相同的一级结构位点，这证实了这两种方法在构象表位定位中的应用。此外，与普马拉病毒感染的早期血清反应的表位与G1的氨基末端区域中的pepspot测定确定。发现所选择的噬菌体克隆与单克隆抗体逃逸突变位点和线性早期表位的相似性（C）1999 Academic Press.
We selected peptide ligands mimicking the surface structure of discontinuous binding sites of Puumala hantavirus-neutralizing monoclonal antibodies from a random 18-amino acid peptide library containing a disulfide bridge in a fixed position and displayed on a filamentous phage. The varying of selection conditions, either by shortening of the association time or by competitive elution with antigen, was crucial for the selection of peptide inserts that could be aligned with the primary sequences of the envelope glycoproteins G1 and G2. Correspondingly, when the envelope glycoprotein sequences were synthesized as overlapping peptides as spots on membrane, the same site in primary structure was found as with phage display, which corroborates the use of the two methods in mapping of conformational epitopes. Also, epitopes reactive with early-phase sera from Puumala virus infection were defined with the pepspot assay in the amino-terminal region of G1. Similarities of the selected phage clones to a monoclonal antibody-escape mutant site and to a linear early-phase epitope were found. (C) 1999 Academic Press.