Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Olin Hall, Ithaca, NY, 14853, USA.
Nancy E. and Peter C. Meinig School of Biomedical Engineering, Cornell University, Weill Hall, Ithaca, NY, 14853, USA.
Nat Commun. 2023 Jan 28;14(1):464. doi: 10.1038/s41467-023-36101-2.
Engineered outer membrane vesicles (OMVs) derived from Gram-negative bacteria are a promising technology for the creation of non-infectious, nanoparticle vaccines against diverse pathogens. However, antigen display on OMVs can be difficult to control and highly variable due to bottlenecks in protein expression and localization to the outer membrane of the host cell, especially for bulky and/or complex antigens. Here, we describe a universal approach for avidin-based vaccine antigen crosslinking (AvidVax) whereby biotinylated antigens are linked to the exterior of OMVs whose surfaces are remodeled with multiple copies of a synthetic antigen-binding protein (SNAP) comprised of an outer membrane scaffold protein fused to a biotin-binding protein. We show that SNAP-OMVs can be readily decorated with a molecularly diverse array of biotinylated subunit antigens, including globular and membrane proteins, glycans and glycoconjugates, haptens, lipids, and short peptides. When the resulting OMV formulations are injected in mice, strong antigen-specific antibody responses are observed that depend on the physical coupling between the antigen and SNAP-OMV delivery vehicle. Overall, these results demonstrate AvidVax as a modular platform that enables rapid and simplified assembly of antigen-studded OMVs for application as vaccines against pathogenic threats.
工程化的革兰氏阴性细菌外膜囊泡（OMVs）是一种有前途的技术，可用于创建针对多种病原体的非感染性纳米颗粒疫苗。然而，由于宿主细胞外膜上的蛋白质表达和定位存在瓶颈，抗原在 OMV 上的展示可能难以控制且高度可变，尤其是对于体积大且/或结构复杂的抗原。在这里，我们描述了一种基于亲和素的疫苗抗原交联（AvidVax）的通用方法，其中生物素化抗原与经过多次修饰的合成抗原结合蛋白（SNAP）的 OMV 表面相连，该 SNAP 由外膜支架蛋白融合到生物素结合蛋白组成。我们表明，SNAP-OMVs 可以很容易地用分子上多样化的生物素化亚单位抗原进行修饰，包括球状和膜蛋白、聚糖和糖缀合物、半抗原、脂质和短肽。当将由此产生的 OMV 制剂注射到小鼠中时，会观察到强烈的抗原特异性抗体反应，这取决于抗原与 SNAP-OMV 递送载体之间的物理偶联。总的来说，这些结果表明 AvidVax 是一种模块化平台，可用于快速简化组装带有抗原的 OMV，作为针对致病威胁的疫苗应用。
