Centre for Immune Regulation, University of Oslo, Oslo, Norway.
PLoS One. 2011 Feb 24;6(2):e14702. doi: 10.1371/journal.pone.0014702.
Phage display is a platform for selection of specific binding molecules and this is a clear-cut motivation for increasing its performance. Polypeptides are normally displayed as fusions to the major coat protein VIII (pVIII), or the minor coat protein III (pIII). Display on other coat proteins such as pVII allows for display of heterologous peptide sequences on the virions in addition to those displayed on pIII and pVIII. In addition, pVII display is an alternative to pIII or pVIII display.
METHODOLOGY/PRINCIPAL FINDINGS: Here we demonstrate how standard pIII or pVIII display phagemids are complemented with a helper phage which supports production of virions that are tagged with octa FLAG, HIS(6) or AviTag on pVII. The periplasmic signal sequence required for pIII and pVIII display, and which has been added to pVII in earlier studies, is omitted altogether.
CONCLUSIONS/SIGNIFICANCE: Tagging on pVII is an important and very useful add-on feature to standard pIII and pVII display. Any phagemid bearing a protein of interest on either pIII or pVIII can be tagged with any of the tags depending simply on choice of helper phage. We show in this paper how such tags may be utilized for immobilization and separation as well as purification and detection of monoclonal and polyclonal phage populations.
噬菌体展示是一种用于筛选特定结合分子的平台，这是提高其性能的明确动机。多肽通常作为主要外壳蛋白 VIII（pVIII）或次要外壳蛋白 III（pIII）的融合体进行展示。在其他外壳蛋白（如 pVII）上展示允许在病毒粒子上展示除了在 pIII 和 pVIII 上展示的那些之外的异源肽序列。此外，pVII 展示是 pIII 或 pVIII 展示的替代方法。
方法/主要发现：在这里，我们展示了如何用辅助噬菌体补充标准的 pIII 或 pVIII 展示噬菌粒，该辅助噬菌体支持在 pVII 上标记八聚 FLAG、HIS（6）或 AviTag 的病毒粒子的生产。先前研究中已添加到 pVII 的用于 pIII 和 pVIII 展示的周质信号序列已完全省略。
结论/意义：在 pVII 上进行标记是标准的 pIII 和 pVII 展示的一个重要且非常有用的附加功能。任何在 pIII 或 pVIII 上带有感兴趣蛋白的噬菌粒都可以根据辅助噬菌体的选择标记任何一种标签。我们在本文中展示了这些标签如何可用于固定和分离以及单克隆和多克隆噬菌体群体的纯化和检测。
