Development of a Recombinant Omicron BA.1 Subunit Vaccine Candidate in Pichia pastoris

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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Main Authors: 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
Format: Article
Language:English
Published: Wiley 2025-01-01
Series:Microbial Biotechnology
Subjects:
COVID‐19 vaccine
omicron BA.1
Pichia pastoris
recombinant protein subunit vaccine
SARS‐CoV‐2
Online Access:https://doi.org/10.1111/1751-7915.70077
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Summary: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.
ISSN:1751-7915

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