Institute of Chemical Sciences and Engineering, School of Basic Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne CH-1015, Switzerland.
Protein Eng Des Sel. 2021 Feb 15;34. doi: 10.1093/protein/gzab018.
Phage display is a powerful technique routinely used for the generation of peptide- or protein-based ligands. The success of phage display selections critically depends on the size and structural diversity of the libraries, but the generation of large libraries remains challenging. In this work, we have succeeded in developing a phage display library comprising around 100 billion different (bi)cyclic peptides and thus more structures than any previously reported cyclic peptide phage display library. Building such a high diversity was achieved by combining a recently reported library cloning technique, based on whole plasmid PCR, with a small plasmid that facilitated bacterial transformation. The library cloned is based on 273 different peptide backbones and thus has a large skeletal diversity. Panning of the peptide repertoire against the important thrombosis target coagulation factor XI enriched high-affinity peptides with long consensus sequences that can only be found if the library diversity is large.
噬菌体展示是一种强大的技术，常用于生成肽或蛋白质基配体。噬菌体展示选择的成功关键取决于文库的大小和结构多样性，但生成大型文库仍然具有挑战性。在这项工作中，我们成功开发了一种噬菌体展示文库，其中包含约 1000 亿个不同的（双）环肽，因此结构比以前报道的任何环状肽噬菌体展示文库都多。通过将基于全质粒 PCR 的最近报道的文库克隆技术与方便细菌转化的小质粒相结合，实现了如此高的多样性。该文库基于 273 种不同的肽骨架构建，因此具有很大的骨架多样性。针对重要的血栓靶点凝血因子 XI 进行的肽库筛选，富集了具有长共有序列的高亲和力肽，如果文库多样性大，只能找到这些序列。
