Bacteriophage MS2 Virus-Like Particles for Peptide Display

描述（由申请人提供）：噬菌体展示是一种非常强大和通用的技术，能够从大量随机产生的肽序列中选择新的结合功能。从足够复杂的文库中，可以通过亲和选择物理分离携带具有实际上任何所需结合活性的肽的噬菌体，并且由于每个颗粒在其基因组中携带用于其自身复制的遗传信息，因此可以在细菌中扩增选择子。本项目的目的是开发一种新的平台，用于在RNA噬菌体MS 2的病毒样颗粒（VLP）上展示肽。我们设想了MS 2 VLP的几种应用，但我们希望特别强调其在疫苗开发中的实用性。它将VLP的有效免疫原性与常规噬菌体展示的亲和选择能力整合到单个平台中。丝状噬菌体是目前应用最广泛的噬菌体展示载体，为表位鉴定提供了一种有效的手段。然而，它们展示的肽通常免疫原性差，因为它们通常不支持密集重复阵列的形成。同时，其他VLP系统允许工程化展示特异性预选表位，但不能进行肽库展示和亲和力选择。我们认为MS 2 VLP将克服这些限制。展示在MS 2 VLP上的肽具有强烈的免疫原性，并且可以被工程化以使编码它们的相同mRNA分子变性，从而使得能够通过RT-PCR回收亲和力选择的序列。此外，MS 2 VLP结构和组装的相对简单性使得可以在体外进行整个迭代选择/扩增过程。这可以更容易地实现高库复杂性，并使自动化成为可能。 公共卫生相关性：该项目旨在开发一种新的平台，用于使用噬菌体MS 2的病毒样颗粒进行肽展示。设想了几种应用，但由于其强大的免疫原性，这些颗粒应该特别适用于疫苗的发现。

DESCRIPTION (provided by applicant): Phage display is an extraordinarily powerful and versatile technology that enables the selection of novel binding functions from large populations of randomly generated peptide sequences. From a sufficiently complex library, phage bearing peptides with practically any desired binding activity can be physically isolated by affinity selection, and, since each particle carries in its genome the genetic information for its own replication, the selectants can be amplified in bacteria. This aim of this project is to develop a new platform for peptide display on virus-like particles (VLPs) of the RNA bacteriophage MS2. We envision several applications for the MS2 VLP, but we wish especially to emphasize its utility for vaccine development. It will integrate into a single platform the potent immunogenicity of a VLP with the affinity selection capability of conventional phage display. Filamentous phages are now the most widely used vehicles for phage display, and provide an efficient means for epitope identification. However, the peptides they display are typically poorly immunogenic, because they do not normally support the formation of dense repetitive arrays. Meanwhile, other VLP systems permit engineered display of specfic pre-selected epitopes, but are incapable of peptide library display and affinity selection. We think MS2 VLPs will overcome these limitations. Peptides displayed on MS2 VLPs are strongly immunogenic, and can be engineered to encapsidate the same mRNA molecule that encodes them, thus enabling recovery of affinity selected sequences by RT-PCR. Further, the comparative simplicity of MS2 VLP structure and assembly makes it possible to conduct the entire iterative selection/amplification process in vitro. This could make it easier to achieve high library complexities, and should make automation possible. PUBLIC HEALTH RELEVANCE: This project aims to develop a new platform for peptide display using virus-like particles of bacteriophage MS2. Several applications are envisioned, but because of their potent immunogenicity, these particles should be especially useful for vaccine discovery.