Bakhshinejad Babak, Ghiasvand Saeedeh
Cluster for Molecular Imaging, Department of Biomedical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark.
Department of Clinical Physiology and Nuclear Medicine, Copenhagen University Hospital-Rigshospitalet, 2100 Copenhagen, Denmark.
Viruses. 2025 Jul 12;17(7):975. doi: 10.3390/v17070975.
Phage display has advanced the discovery of peptides that selectively bind to a wide variety of cell surface molecules, offering new modalities to modulate disease-related protein-protein interactions (PPIs). These cell-binding peptides occupy a unique pharmaceutical space between small molecules and large biologics, and their growing popularity has opened up new avenues for targeting cell surface proteins that were previously considered undruggable. This work provides an overview of methods for identifying cell-selective peptides using phage display combinatorial libraries, covering in vitro, ex vivo, and in vivo biopanning approaches. It addresses key considerations in library design, including the peptide conformation (linear vs. cyclic) and length, and highlights examples of clinically approved peptides developed through phage display. It also discusses the on-phage chemical cyclization of peptides to overcome the limitations of genetically encoded disulfide bridges and emphasizes advances in combining next-generation sequencing (NGS) with phage display to improve peptide selection and analysis workflows. Furthermore, due to the often suboptimal binding affinity of peptides identified in phage display selections, this article discusses affinity maturation techniques, including random mutagenesis and rational design through structure-activity relationship (SAR) studies to optimize initial peptide candidates. By integrating these developments, this review outlines practical strategies and future directions for harnessing phage display in targeting challenging cell surface proteins.
噬菌体展示技术推动了对能选择性结合多种细胞表面分子的肽的发现，为调节疾病相关的蛋白质-蛋白质相互作用（PPI）提供了新的方式。这些细胞结合肽在小分子和大分子生物制剂之间占据着独特的药物空间，并且它们越来越受欢迎，为靶向以前被认为不可成药的细胞表面蛋白开辟了新途径。这项工作概述了使用噬菌体展示组合文库鉴定细胞选择性肽的方法，涵盖体外、离体和体内淘选方法。它阐述了文库设计中的关键考虑因素，包括肽的构象（线性与环状）和长度，并突出了通过噬菌体展示开发的临床批准肽的实例。它还讨论了肽的噬菌体上化学环化以克服基因编码二硫键的局限性，并强调了将下一代测序（NGS）与噬菌体展示相结合以改进肽选择和分析工作流程的进展。此外，由于在噬菌体展示筛选中鉴定出的肽的结合亲和力往往不理想，本文讨论了亲和力成熟技术，包括随机诱变和通过构效关系（SAR）研究进行理性设计以优化初始肽候选物。通过整合这些进展，本综述概述了利用噬菌体展示靶向具有挑战性的细胞表面蛋白的实用策略和未来方向。
