Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Viale F. Stagno d'Alcontres 31, 98166 Messina, Italy.
Pasteur Institute, University of Paris, Synthetic Biology, 75015 Paris, France.
Cells. 2024 Oct 14;13(20):1696. doi: 10.3390/cells13201696.
Phage display is widely used in biomedical research. One of the great advantages of phage display is the specificity of the connection of a foreign peptide exposed outside the capsid to the intended target. Secondary detection systems, which are often laborious and costly, are required to identify and quantify the peptide/target interaction. In this study, we generated a novel dual-display phage to facilitate the detection and quantification of the peptide/target interaction. First, we generated a biotin-tagged phage by adding a small biotin-accepting peptide (sBT) to gene-3 of the M13K07 helper phage. Subsequently, we enhanced the M13K07 biotin-tagged phage by incorporating a selective peptide on gene-8, which is then exposed to the phage capsid. The exposed peptide acts as a probe to bind to a selective molecular target, whose interaction can be readily visualized thanks to the biotinylated phage. Our versatile dual-display phage exhibits high flexibility; by swapping the displayed peptide/probe, one can change the phage target while retaining the sBT gene in-frame with the pIII. We expect the generated biotin-tagged dual phages to be used as a multifunctional probe to couple with several streptavidin-biotin-based systems.
噬菌体展示技术广泛应用于生物医学研究。噬菌体展示的一个巨大优势是，外壳蛋白外暴露的外源肽与预期目标的连接具有特异性。通常需要二级检测系统来识别和定量肽/靶相互作用，但该系统既繁琐又昂贵。在本研究中，我们生成了一种新型双展示噬菌体，以促进肽/靶相互作用的检测和定量。首先，我们通过在 M13K07 辅助噬菌体的基因 3 中添加一个小的生物素接受肽 (sBT) 来生成生物素标记的噬菌体。随后，我们在基因 8 上整合了一个选择性肽，增强了 M13K07 生物素标记的噬菌体，然后将其暴露在噬菌体外壳上。暴露的肽作为探针与选择性分子靶标结合，由于生物素化的噬菌体，其相互作用可以很容易地可视化。我们灵活的双展示噬菌体具有很高的灵活性；通过交换展示的肽/探针，可以在保留 sBT 基因与 pIII 框内的情况下改变噬菌体的靶标。我们期望生成的生物素标记的双噬菌体能够用作多功能探针，与几个链霉亲和素-生物素为基础的系统相结合。
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