Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidate
Abstract 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...
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Nature Portfolio
2025-05-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-025-01242-5 |
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| author | Ozi Jumadila Muhammad Dzul Fakhri Adam Darsono Fernita Puspasari Sari Dewi Kurniasih Fifi Fitriyah Masduki Keni Vidilaseris Ihsanawati Ihsanawati Anita Artarini Marselina Irasonia Tan Ernawati Arifin Giri-Rachman Dessy Natalia |
| author_facet | Ozi Jumadila Muhammad Dzul Fakhri Adam Darsono Fernita Puspasari Sari Dewi Kurniasih Fifi Fitriyah Masduki Keni Vidilaseris Ihsanawati Ihsanawati Anita Artarini Marselina Irasonia Tan Ernawati Arifin Giri-Rachman Dessy Natalia |
| author_sort | Ozi Jumadila |
| collection | DOAJ |
| description | Abstract 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. |
| format | Article |
| id | doaj-art-8ad37908ece54c6d93deeb2a089965b0 |
| institution | OA Journals |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-8ad37908ece54c6d93deeb2a089965b02025-08-20T02:00:08ZengNature PortfolioScientific Reports2045-23222025-05-0115111510.1038/s41598-025-01242-5Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidateOzi Jumadila0Muhammad Dzul Fakhri1Adam Darsono2Fernita Puspasari3Sari Dewi Kurniasih4Fifi Fitriyah Masduki5Keni Vidilaseris6Ihsanawati Ihsanawati7Anita Artarini8Marselina Irasonia Tan9Ernawati Arifin Giri-Rachman10Dessy Natalia11Biochemistry and Biomolecular Engineering Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi BandungBiochemistry and Biomolecular Engineering Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi BandungSchool of Life Sciences and Technology, Institut Teknologi BandungBiochemistry and Biomolecular Engineering Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi BandungBiochemistry and Biomolecular Engineering Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi BandungBiochemistry and Biomolecular Engineering Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi BandungThe Molecular and Integrative Biosciences Research Programme (MIBS), Faculty of Biological and Environmental Sciences, University of HelsinkiBiochemistry and Biomolecular Engineering Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi BandungLaboratory of Pharmaceutical Biotechnology, School of Pharmacy, Institut Teknologi BandungSchool of Life Sciences and Technology, Institut Teknologi BandungSchool of Life Sciences and Technology, Institut Teknologi BandungBiochemistry and Biomolecular Engineering Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi BandungAbstract 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.https://doi.org/10.1038/s41598-025-01242-5COVID-19Multimeric proteinRBDSubunit vaccineβ-annulus |
| spellingShingle | Ozi Jumadila Muhammad Dzul Fakhri Adam Darsono Fernita Puspasari Sari Dewi Kurniasih Fifi Fitriyah Masduki Keni Vidilaseris Ihsanawati Ihsanawati Anita Artarini Marselina Irasonia Tan Ernawati Arifin Giri-Rachman Dessy Natalia Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidate Scientific Reports COVID-19 Multimeric protein RBD Subunit vaccine β-annulus |
| title | Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidate |
| title_full | Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidate |
| title_fullStr | Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidate |
| title_full_unstemmed | Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidate |
| title_short | Development of a self-assembling multimeric Bann-RBD fusion protein in Pichia pastoris as a potential COVID-19 vaccine candidate |
| title_sort | development of a self assembling multimeric bann rbd fusion protein in pichia pastoris as a potential covid 19 vaccine candidate |
| topic | COVID-19 Multimeric protein RBD Subunit vaccine β-annulus |
| url | https://doi.org/10.1038/s41598-025-01242-5 |
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