Bakhshinejad Babak, Kjaer Andreas
Cluster for Molecular Imaging, Department of Biomedical Sciences, University of Copenhagen, Copenhagen, Denmark.
Department of Clinical Physiology and Nuclear Medicine, Copenhagen University Hospital-Rigshospitalet, Copenhagen, Denmark.
Front Microbiol. 2025 Jun 4;16:1571679. doi: 10.3389/fmicb.2025.1571679. eCollection 2025.
Over the recent decades, phage display has been used successfully to identify a variety of peptides with diagnostic and therapeutic applications. Despite the significant role of this technology in the pharmaceutical industry, the affinity selection of phage display peptide libraries through biopanning suffers from some limitations. The most significant drawback of phage display is the undesirable enrichment and isolation of phages whose displayed peptides have no binding affinity toward the target. Phages with high amplification rates constitute the most important category of non-specific binders. Amplification, which aims to increase the copy number of phages displaying target-specific peptides, acts like a double-edged blade and can also make a major contribution to the target-unrelated enrichment of non-specific binders, leading to compositional bias in the sequence content of the biopanning output. The cutting-edge breakthroughs fueled by the integration of next-generation sequencing (NGS) into phage display have led researchers to gain a deeper understanding of the information content of the phage population recovered from biopanning and how its peptide content changes during further rounds of selection and amplification. This body of vastly increasing information has shed more light on the complications encountered during library selection and opened new perspectives to obtain in-depth insights into amplification-associated bias in the selected phage display libraries, analyze biopanning data more rigorously, and devise more optimal protocols for phage display selections. This knowledge can finally provide a solid foundation for discovering promising target-specific binders in the evolutionary selection of phage display libraries.
在最近几十年里，噬菌体展示已成功用于鉴定多种具有诊断和治疗应用的肽。尽管这项技术在制药行业发挥了重要作用，但通过生物淘选对噬菌体展示肽库进行亲和力选择仍存在一些局限性。噬菌体展示最显著的缺点是，展示的肽对靶标没有结合亲和力的噬菌体出现不良富集和分离。扩增率高的噬菌体是非特异性结合物的最重要类别。扩增旨在增加展示靶标特异性肽的噬菌体的拷贝数，就像一把双刃剑，也会对非特异性结合物与靶标无关的富集产生重大影响，导致生物淘选输出序列内容的组成偏差。将下一代测序（NGS）整合到噬菌体展示中所带来的前沿突破，使研究人员能够更深入地了解从生物淘选中回收的噬菌体群体的信息内容，以及其肽含量在进一步的选择和扩增轮次中是如何变化的。这大量不断增加的信息，让人们更加清楚地了解文库选择过程中遇到的复杂情况，并为深入了解所选噬菌体展示文库中与扩增相关的偏差、更严格地分析生物淘选数据以及设计更优化的噬菌体展示选择方案开辟了新的视角。这些知识最终可为在噬菌体展示文库的进化选择中发现有前景的靶标特异性结合物提供坚实基础。
