PDF(Pubmed)
Abstract：
Virus-like particles (VLPs) are nanostructures with the potential to present heterologous peptides at high density, thereby triggering heightened immunogenicity. RNA bacteriophage MS2 VLPs are a compelling delivery platform among them. However, a notable hurdle arises from the immune response toward MS2 coat protein, swiftly eliminating subsequent vaccinations via the same vector. Although larger inserts effectively mask carrier epitopes, current research predominantly focuses on displaying short conserved peptides (<30 aa). A systematic evaluation regarding the deterministic ability of MS2 VLPs as a platform for presenting heterologous peptides remains a gap. In light of this, we employed the \"single-chain dimer\" paradigm to scrutinize the tolerance of MS2 VLPs for peptide/protein insertions. The results unveiled functional MS2 VLP assembly solely for inserts smaller than 91 aa. Particularly noteworthy is the largest insertion achieved on the MS2 VLPs to date: the RNA helicase A (RHA) dsRNA-binding domains (dsRBD1). Attempts to introduce additional linkers or empty coat subunits fail to augment the expression level or assembly of the MS2 VLPs displaying dsRBD1, affirming 91 aa as the upper threshold for exogenous protein presentation. By illuminating the precise confines of MS2 VLPs in accommodating distinct peptide lengths, our study informs the selection of appropriate peptide and protein dimensions. This revelation not only underscores the scope of MS2 VLPs but also establishes a pivotal reference point, facilitating the strategic manipulation of MS2 VLPs to design next-generation epitope/antibody-based therapeutics.
摘要：
病毒样颗粒（VLP）是纳米结构，有可能以高密度呈现异源肽，从而引发增强的免疫原性。RNA噬菌体MS2VLP是其中一个引人注目的递送平台。然而,一个值得注意的障碍来自对MS2外壳蛋白的免疫反应，通过相同的载体迅速消除后续疫苗接种。虽然更大的插入物有效地掩盖了载体表位，目前的研究主要集中在展示短保守肽（<30aa）。关于MS2VLP作为呈递异源肽的平台的确定性能力的系统评估仍然是空白。鉴于此,我们采用“单链二聚体”范式来审查MS2VLP对肽/蛋白质插入的耐受性。结果揭示了仅适用于小于91aa的插入件的功能性MS2VLP组件。特别值得注意的是迄今为止在MS2VLP上实现的最大插入:RNA解旋酶A(RHA)dsRNA结合结构域(dsRBD1)。尝试引入其他接头或空外壳亚基无法增加显示dsRBD1的MS2VLP的表达水平或组装，确认91aa为外源蛋白呈递的上限阈值。通过阐明MS2VLP在适应不同肽长度方面的精确界限，我们的研究为选择合适的肽和蛋白质尺寸提供了信息。这一启示不仅强调了MS2VLP的范围，而且建立了一个关键的参考点，促进MS2VLP的战略操作，以设计下一代基于表位/抗体的疗法。
