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首页> 外文期刊>Journal of Molecular Biology >Peptide epitope identification by affinity selection on bacteriophage MS2 virus-like particles.
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Peptide epitope identification by affinity selection on bacteriophage MS2 virus-like particles.

机译：通过对噬菌体MS2病毒样颗粒的亲和力选择来鉴定肽表位。

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摘要

Filamentous phages are now the most widely used vehicles for phage display and provide efficient means for epitope identification. However, the peptides they display are not very immunogenic because they normally fail to present foreign epitopes at the very high densities required for efficient B-cell activation. Meanwhile, systems based on virus-like particles (VLPs) permit the engineered high-density display of specific epitopes but are incapable of peptide library display and affinity selection. We developed a new peptide display platform based on VLPs of the RNA bacteriophage MS2. It combines the high immunogenicity of MS2 VLPs with the affinity selection capabilities of other phage display systems. Here, we describe plasmid vectors that facilitate the construction of high-complexity random sequence peptide libraries on MS2 VLPs and that allow control of the stringency of affinity selection through the manipulation of display valency. We used the system to identify epitopes for several previously characterized monoclonal antibody targets and showed that the VLPs thus obtained elicit antibodies in mice whose activities mimic those of the selecting antibodies.
机译：丝状噬菌体现在是用于噬菌体展示的最广泛使用的载体，并提供有效的方法来鉴定表位。然而，它们展示的肽不是很强的免疫原性，因为它们通常不能以有效B细胞活化所需的很高密度呈递外来抗原决定簇。同时，基于病毒样颗粒（VLP）的系统允许对特定表位进行工程化的高密度展示，但无法进行肽库展示和亲和力选择。我们开发了基于RNA噬菌体MS2的VLP的新肽展示平台。它结合了MS2 VLP的高免疫原性和其他噬菌体展示系统的亲和力选择功能。在这里，我们描述了质粒载体，该质粒载体有助于在MS2 VLP上构建高复杂度的随机序列肽库，并允许通过操纵展示价来控制亲和力选择的严格性。我们使用该系统为几个先前表征的单克隆抗体靶标鉴定了抗原决定簇，并表明，VLP因此在小鼠中诱发了抗体，这些小鼠的活性与选择抗体的活性类似。

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