tetano
Editor, Senior Moderator
Sci Rep
. 2025 May 27;15(1):18467.
doi: 10.1038/s41598-025-01242-5. Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidate
Ozi Jumadila[SUP] 1 [/SUP], Muhammad Dzul Fakhri[SUP] 1 [/SUP], Adam Darsono[SUP] 2 [/SUP], Fernita Puspasari[SUP] 1 [/SUP], Sari Dewi Kurniasih[SUP] 1 [/SUP], Fifi Fitriyah Masduki[SUP] 1 3 [/SUP], Keni Vidilaseris[SUP] 4 [/SUP], Ihsanawati Ihsanawati[SUP] 1 [/SUP], Anita Artarini[SUP] 5 3 [/SUP], Marselina Irasonia Tan[SUP] 2 3 [/SUP], Ernawati Arifin Giri-Rachman[SUP] 2 3 [/SUP], Dessy Natalia[SUP] 6 7 [/SUP]
Affiliations
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.
Keywords: COVID-19; Multimeric protein; RBD; Subunit vaccine; β-annulus.
. 2025 May 27;15(1):18467.
doi: 10.1038/s41598-025-01242-5. Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidate
Ozi Jumadila[SUP] 1 [/SUP], Muhammad Dzul Fakhri[SUP] 1 [/SUP], Adam Darsono[SUP] 2 [/SUP], Fernita Puspasari[SUP] 1 [/SUP], Sari Dewi Kurniasih[SUP] 1 [/SUP], Fifi Fitriyah Masduki[SUP] 1 3 [/SUP], Keni Vidilaseris[SUP] 4 [/SUP], Ihsanawati Ihsanawati[SUP] 1 [/SUP], Anita Artarini[SUP] 5 3 [/SUP], Marselina Irasonia Tan[SUP] 2 3 [/SUP], Ernawati Arifin Giri-Rachman[SUP] 2 3 [/SUP], Dessy Natalia[SUP] 6 7 [/SUP]
Affiliations
- PMID: 40425664
- PMCID: PMC12117094
- 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.
Keywords: COVID-19; Multimeric protein; RBD; Subunit vaccine; β-annulus.