Jung Helen S, Bikash Chowdhury Raihan, Tao Kevin L, Lai Jonathan R
Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York 10461, United States.
ACS Omega. 2025 Jun 2;10(23):25068-25075. doi: 10.1021/acsomega.5c03059. eCollection 2025 Jun 17.
The presentation of antigens on self-assembling nanoparticles is a promising approach for augmenting the B-cell response of vaccines. In many cases, the nanoparticle and antigen are recombinantly expressed and purified separately prior to conjugation, requiring multiple steps that may be inefficient for large-scale production. Here, we describe a genetically encodable method and "one-pot" production of nanoparticle immunogens from mammalian cells. Our approach relies on the assembly of the nanoparticle-antigen conjugate, between protein nanoparticles bearing SpyCatcher003 and antigen fused to SpyTag003, using simple cotransfection in mammalian cells, which can then be purified to homogeneity in a single step. We demonstrate this method with an antigen based on Zika virus E glycoprotein domain III ("rsZDIII-2.39") and compare the biochemical properties and immunogenicity in mice relative to genetic fusions. This work provides a framework for streamlining immunogen production and raises the possibility of employing such approaches for encoding nanoparticle immunogens as nucleic acid vaccines.
在自组装纳米颗粒上呈递抗原是增强疫苗B细胞反应的一种有前景的方法。在许多情况下，纳米颗粒和抗原在缀合之前需分别进行重组表达和纯化，这需要多个步骤，对于大规模生产而言可能效率低下。在此，我们描述了一种可遗传编码的方法以及从哺乳动物细胞中“一锅法”生产纳米颗粒免疫原。我们的方法依赖于在携带SpyCatcher003的蛋白质纳米颗粒与融合有SpyTag003的抗原之间组装纳米颗粒 - 抗原缀合物，通过在哺乳动物细胞中进行简单的共转染来实现，然后可在单个步骤中纯化至同质。我们用基于寨卡病毒E糖蛋白结构域III（“rsZDIII - 2.39”）的抗原展示了该方法，并比较了其与基因融合物相比在小鼠体内的生化特性和免疫原性。这项工作为简化免疫原生产提供了一个框架，并增加了将此类方法用于编码纳米颗粒免疫原作为核酸疫苗的可能性。
