Hishot display - a new combinatorial display for obtaining target-recognizing peptides

显示技术是在不使用免疫动物的情况下分离目标识别肽的程序。在这项研究中，我们描述了一种新的显示方法，称为Hishot显示，利用大肠杆菌和表达质粒分离目标识别肽。这种展示方法是基于在细菌中形成多组氨酸（His）标记的包含随机序列的肽和包含针对His标签的RNA适体的肽编码mRNA之间的复合物。利用这种显示方法和由VpreB和Ig重链可变结构域组成的替代单链可变片段合成库，可以分离出特异性识别特定靶标的克隆。获得的克隆可纯化为大肠杆菌产生的可溶性蛋白。Hishot显示将是一种有用的方法来增加显示技术的曲目。

Display technologies are procedures used for isolating target-recognizing peptides without using immunized animals. In this study, we describe a new display method, named Hishot display, that uses Escherichia coli and an expression plasmid to isolate target-recognizing peptides. This display method is based on the formation, in bacteria, of complexes between a polyhistidine (His)-tagged peptide including random sequences and the peptide-encoding mRNA including an RNA aptamer against the His-tag. Using this display method and a synthetic library of surrogate single-chain variable fragments consisting of VpreB and Ig heavy-chain variable domains, it was possible to isolate clones that could specifically recognize a particular target. The obtained clones could be purified as soluble protein produced by E. coli. The Hishot display will be a useful method to add to the repertoire of display technologies.