Department of Biological Sciences, Michigan Technological University, Houghton, MI, 49931, USA.
Department of Chemical Engineering, Michigan Technological University, Houghton, MI, 49931, USA.
Protein Expr Purif. 2021 Nov;187:105932. doi: 10.1016/j.pep.2021.105932. Epub 2021 Jun 29.
Virus-like particles (VLPs) have the potential to be used as display platforms to develop vaccines against infectious and non-infectious agents. However, most VLPs used as vaccine display platforms are derived from viruses that infect humans; unfortunately, most humans already have pre-existing antibodies against these platforms and thus, the immunogenicity of these vaccines may be compromised. VLP platforms derived from viruses that infect bacteria (bacteriophages), especially bacteriophages that infect bacteria, which do not colonize humans are less likely to have pre-existing antibodies against the platforms in the human population. In this study, we assessed whether two putative coat proteins (ORF13 and ORF14) derived from a thermophilic bacteriophage (ΦIN93) can be expressed and purified from a mesophilic bacterium such as E. coli. We also assessed whether expressed coat proteins can assemble to form VLPs. Truncated versions of ORF13 and ORF14 were successfully co-expressed in bacteria; the co-expressed truncated proteins formed oval structures that look like VLPs, but their sizes were less than those of an authentic ΦIN93 virus.
病毒样颗粒 (VLPs) 具有作为展示平台开发针对传染性和非传染性制剂疫苗的潜力。然而,大多数用作疫苗展示平台的 VLPs 都源自感染人类的病毒;不幸的是,大多数人类已经对这些平台产生了预先存在的抗体,因此这些疫苗的免疫原性可能会受到影响。源自感染细菌的病毒(噬菌体)的 VLP 平台,特别是感染不会定植于人类的细菌的噬菌体,不太可能在人类群体中对平台预先存在抗体。在这项研究中,我们评估了来自嗜热噬菌体(ΦIN93)的两个假定衣壳蛋白(ORF13 和 ORF14)是否可以从诸如大肠杆菌等嗜温细菌中表达和纯化。我们还评估了表达的衣壳蛋白是否可以组装形成 VLPs。ORF13 和 ORF14 的截断版本在细菌中成功共表达;共表达的截断蛋白形成了看起来像 VLP 的椭圆形结构,但它们的大小小于真正的 ΦIN93 病毒。