Bacteriophage MS2 Virus-Like Particles for Peptide Display

描述（由申请人提供）：噬菌体展示是一项非常强大和通用的技术，可以从大量随机生成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出具有几乎任何所需结合活性的噬菌体携带肽，并且由于每个颗粒在其基因组中携带用于自身复制的遗传信息，因此选择剂可以在细菌中扩增。本项目旨在为RNA噬菌体MS2的病毒样颗粒（vlp）上的肽展示开发一个新的平台。我们设想了MS2 VLP的几种应用，但我们希望特别强调它对疫苗开发的效用。它将把VLP的强大免疫原性与传统噬菌体展示的亲和选择能力整合到一个单一的平台中。丝状噬菌体是目前应用最广泛的噬菌体展示载体，为噬菌体表位鉴定提供了一种有效的手段。然而，它们显示的肽通常是免疫原性差的，因为它们通常不支持密集重复阵列的形成。同时，其他VLP系统允许工程显示特定的预先选择的表位，但不能显示肽库和亲和力选择。我们认为MS2 vlp将克服这些限制。MS2 VLPs上显示的肽具有很强的免疫原性，可以被设计成封装编码它们的相同mRNA分子，从而可以通过RT-PCR恢复亲和选择的序列。此外，MS2 VLP的结构和组装相对简单，使得在体外进行整个迭代选择/扩增过程成为可能。这可以使实现高库复杂性变得更容易，并且应该使自动化成为可能。公共卫生相关性：本项目旨在利用噬菌体MS2的病毒样颗粒开发一个新的肽展示平台。设想了几种应用，但由于其强大的免疫原性，这些颗粒应特别有助于疫苗的发现。

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.