Development of a Recombinant Omicron BA.1 Subunit Vaccine Candidate in Pichia pastoris
ABSTRACT Low‐cost and safe vaccines are needed to fill the vaccine inequity gap for future pandemics. Pichia pastoris is an ideal expression system for recombinant protein production due to its cost‐effective and easy‐to‐scale‐up process. Here, we developed a next‐generation SARS‐CoV2 Omicron BA.1‐b...
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| Format: | Article |
| Language: | English |
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Wiley
2025-01-01
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| Series: | Microbial Biotechnology |
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| Online Access: | https://doi.org/10.1111/1751-7915.70077 |
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| author | Sibel Kalyoncu Dogu Sayili Ayca Zeybek Kuyucu Hakan Soyturk Seyda Gullu Busra Ersayan Ibrahim Oguzhan Tarman Mehmet Ender Avci Olcay Mert Umut Haskok Ege Tekin Huseyin Akinturk Ridvan Orkut Aysegul Demirtas Idil Tilmensagir Ceren Ulker Bilgi Gungor Mehmet Inan |
| author_facet | Sibel Kalyoncu Dogu Sayili Ayca Zeybek Kuyucu Hakan Soyturk Seyda Gullu Busra Ersayan Ibrahim Oguzhan Tarman Mehmet Ender Avci Olcay Mert Umut Haskok Ege Tekin Huseyin Akinturk Ridvan Orkut Aysegul Demirtas Idil Tilmensagir Ceren Ulker Bilgi Gungor Mehmet Inan |
| author_sort | Sibel Kalyoncu |
| collection | DOAJ |
| description | ABSTRACT Low‐cost and safe vaccines are needed to fill the vaccine inequity gap for future pandemics. Pichia pastoris is an ideal expression system for recombinant protein production due to its cost‐effective and easy‐to‐scale‐up process. Here, we developed a next‐generation SARS‐CoV2 Omicron BA.1‐based recombinant vaccine candidate expressed in P. pastoris. The receptor binding domain of Omicron BA.1 spike protein (RBD‐Omicron) was produced at 0.35 g/L in supernatant. With a 60% recovery after two‐step purification, RBD‐Omicron showed 99% purity. After in vitro characterisation of purified RBD‐Omicron via chromatography, mass spectrometry, calorimetry and surface plasmon resonance‐based methods, it was injected into mice for immunization studies. Three different doses of Alum and CpG adjuvanted RBD‐Omicron were investigated and 10 μg RBD‐Omicron gave the highest antigenicity. After two doses of vaccination, IgG titers in mice serum reached to more than 106. These serum antibodies also recognized earlier (Delta Plus: B.1.617.2) and later (Eris: EG.5, Pirola: BA.2.86) SARS‐CoV2 variants. The long‐term immunological response in mice was measured by analyzing serum antibody titers and T‐cell response of splenocytes after 60 weeks. Interestingly, IgG titers and Th1 response were significantly high even after a year. Omicron subvariants are dominantly circulating in the world, so Omicron sub‐lineage‐based vaccines can be used for future pandemics. The RBD‐Omicron‐based vaccine candidate developed in this study is suitable for technology transfer and transition into the clinic. |
| format | Article |
| id | doaj-art-2fda2c17432140cd9205bfb1dbd355ba |
| institution | Kabale University |
| issn | 1751-7915 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Microbial Biotechnology |
| spelling | doaj-art-2fda2c17432140cd9205bfb1dbd355ba2025-01-31T06:26:35ZengWileyMicrobial Biotechnology1751-79152025-01-01181n/an/a10.1111/1751-7915.70077Development of a Recombinant Omicron BA.1 Subunit Vaccine Candidate in Pichia pastorisSibel Kalyoncu0Dogu Sayili1Ayca Zeybek Kuyucu2Hakan Soyturk3Seyda Gullu4Busra Ersayan5Ibrahim Oguzhan Tarman6Mehmet Ender Avci7Olcay Mert8Umut Haskok9Ege Tekin10Huseyin Akinturk11Ridvan Orkut12Aysegul Demirtas13Idil Tilmensagir14Ceren Ulker15Bilgi Gungor16Mehmet Inan17Izmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyIzmir Biomedicine and Genome Center Izmir TurkeyABSTRACT Low‐cost and safe vaccines are needed to fill the vaccine inequity gap for future pandemics. Pichia pastoris is an ideal expression system for recombinant protein production due to its cost‐effective and easy‐to‐scale‐up process. Here, we developed a next‐generation SARS‐CoV2 Omicron BA.1‐based recombinant vaccine candidate expressed in P. pastoris. The receptor binding domain of Omicron BA.1 spike protein (RBD‐Omicron) was produced at 0.35 g/L in supernatant. With a 60% recovery after two‐step purification, RBD‐Omicron showed 99% purity. After in vitro characterisation of purified RBD‐Omicron via chromatography, mass spectrometry, calorimetry and surface plasmon resonance‐based methods, it was injected into mice for immunization studies. Three different doses of Alum and CpG adjuvanted RBD‐Omicron were investigated and 10 μg RBD‐Omicron gave the highest antigenicity. After two doses of vaccination, IgG titers in mice serum reached to more than 106. These serum antibodies also recognized earlier (Delta Plus: B.1.617.2) and later (Eris: EG.5, Pirola: BA.2.86) SARS‐CoV2 variants. The long‐term immunological response in mice was measured by analyzing serum antibody titers and T‐cell response of splenocytes after 60 weeks. Interestingly, IgG titers and Th1 response were significantly high even after a year. Omicron subvariants are dominantly circulating in the world, so Omicron sub‐lineage‐based vaccines can be used for future pandemics. The RBD‐Omicron‐based vaccine candidate developed in this study is suitable for technology transfer and transition into the clinic.https://doi.org/10.1111/1751-7915.70077COVID‐19 vaccineomicron BA.1Pichia pastorisrecombinant protein subunit vaccineSARS‐CoV‐2 |
| spellingShingle | Sibel Kalyoncu Dogu Sayili Ayca Zeybek Kuyucu Hakan Soyturk Seyda Gullu Busra Ersayan Ibrahim Oguzhan Tarman Mehmet Ender Avci Olcay Mert Umut Haskok Ege Tekin Huseyin Akinturk Ridvan Orkut Aysegul Demirtas Idil Tilmensagir Ceren Ulker Bilgi Gungor Mehmet Inan Development of a Recombinant Omicron BA.1 Subunit Vaccine Candidate in Pichia pastoris Microbial Biotechnology COVID‐19 vaccine omicron BA.1 Pichia pastoris recombinant protein subunit vaccine SARS‐CoV‐2 |
| title | Development of a Recombinant Omicron BA.1 Subunit Vaccine Candidate in Pichia pastoris |
| title_full | Development of a Recombinant Omicron BA.1 Subunit Vaccine Candidate in Pichia pastoris |
| title_fullStr | Development of a Recombinant Omicron BA.1 Subunit Vaccine Candidate in Pichia pastoris |
| title_full_unstemmed | Development of a Recombinant Omicron BA.1 Subunit Vaccine Candidate in Pichia pastoris |
| title_short | Development of a Recombinant Omicron BA.1 Subunit Vaccine Candidate in Pichia pastoris |
| title_sort | development of a recombinant omicron ba 1 subunit vaccine candidate in pichia pastoris |
| topic | COVID‐19 vaccine omicron BA.1 Pichia pastoris recombinant protein subunit vaccine SARS‐CoV‐2 |
| url | https://doi.org/10.1111/1751-7915.70077 |
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