Jumadila Ozi, Fakhri Muhammad Dzul, Darsono Adam, Puspasari Fernita, Kurniasih Sari Dewi, Masduki Fifi Fitriyah, Vidilaseris Keni, Ihsanawati Ihsanawati, Artarini Anita, Tan Marselina Irasonia, Giri-Rachman Ernawati Arifin, Natalia Dessy
Biochemistry and Biomolecular Engineering Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Jl. Ganesa No. 10, Bandung, 40132, Indonesia.
School of Life Sciences and Technology, Institut Teknologi Bandung, Jl. Ganesa No. 10, Bandung, 40132, Indonesia.
Sci Rep. 2025 May 27;15(1):18467. doi: 10.1038/s41598-025-01242-5.
The development of an affordable and accessible vaccine platform is essential for achieving global and long-term protection against COVID-19 and other emerging viral diseases. In this study, we developed a multimeric fusion protein comprising the SARS-CoV-2 receptor-binding domain (RBD) and the β-annulus (Bann) from the tomato bushy stunt virus (TBSV) as a potential subunit vaccine candidate. Molecular modeling of Bann-RBD revealed a 60-mer structure with the RBD displayed on its outer surface. The Bann-RBD gene was constructed and overexpressed in Pichia pastoris X-33. SDS-PAGE analysis of the purified Bann-RBD showed a protein band at 45 kDa, corresponding to monomeric glycosylated Bann-RBD. Peptide mapping analysis using LC-MS/MS confirmed that the expressed Bann-RBD was consistent with the designed protein fusion. The Bann-RBD protein was observed to spontaneously self-assemble into spherical nanocapsids with a diameter of approximately 50 nm. Antigenicity studies demonstrated that the purified Bann-RBD was strongly recognised by monoclonal human anti-SARS-CoV-2 spike-S1 IgG antibodies. Immunogenicity studies revealed that Bann-RBD elicited a robust humoral immune response in BALB/c mice, generating potent neutralising antibodies. Collectively, these findings suggest that the recombinant Bann-RBD produced in Pichia pastoris X-33 is a promising candidate for the development of a COVID-19 vaccine.
开发一种价格合理且易于获取的疫苗平台对于实现全球长期预防新冠病毒（COVID-19）及其他新兴病毒性疾病至关重要。在本研究中，我们构建了一种多聚体融合蛋白，其包含严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的受体结合域（RBD）和来自番茄丛矮病毒（TBSV）的β-环（Bann），作为一种潜在的亚单位疫苗候选物。Bann-RBD的分子建模显示其为一种60聚体结构，RBD展示在其外表面。构建了Bann-RBD基因并在毕赤酵母X-33中进行过表达。对纯化后的Bann-RBD进行的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析显示，在45 kDa处有一条蛋白条带，对应于单体糖基化的Bann-RBD。使用液相色谱-串联质谱（LC-MS/MS）进行的肽图分析证实，表达的Bann-RBD与设计的蛋白融合体一致。观察到Bann-RBD蛋白可自发自组装成直径约为50 nm的球形纳米衣壳。抗原性研究表明，纯化后的Bann-RBD能被人抗SARS-CoV-2刺突S1 IgG单克隆抗体强烈识别。免疫原性研究显示，Bann-RBD在BALB/c小鼠中引发了强烈的体液免疫反应，产生了强效中和抗体。总体而言，这些发现表明，在毕赤酵母X-33中产生的重组Bann-RBD是开发COVID-19疫苗的一个有前景的候选物。
