Scientific Reports (Mar 2023)
Process development for an effective COVID-19 vaccine candidate harboring recombinant SARS-CoV-2 delta plus receptor binding domain produced by Pichia pastoris
- Sibel Kalyoncu,
- Semiramis Yilmaz,
- Ayca Zeybek Kuyucu,
- Dogu Sayili,
- Olcay Mert,
- Hakan Soyturk,
- Seyda Gullu,
- Huseyin Akinturk,
- Erhan Citak,
- Merve Arslan,
- Melda Guray Taskinarda,
- Ibrahim Oguzhan Tarman,
- Gizem Yilmazer Altun,
- Ceren Ozer,
- Ridvan Orkut,
- Aysegul Demirtas,
- Idil Tilmensagir,
- Umur Keles,
- Ceren Ulker,
- Gizem Aralan,
- Yavuz Mercan,
- Muge Ozkan,
- Hasan Onur Caglar,
- Gizem Arik,
- Mehmet Can Ucar,
- Muzaffer Yildirim,
- Tugce Canavar Yildirim,
- Dilara Karadag,
- Erhan Bal,
- Aybike Erdogan,
- Serif Senturk,
- Serdar Uzar,
- Hakan Enul,
- Cumhur Adiay,
- Fahriye Sarac,
- Arzu Tas Ekiz,
- Irem Abaci,
- Ozge Aksoy,
- Hivda Ulbegi Polat,
- Saban Tekin,
- Stefan Dimitrov,
- Aykut Ozkul,
- Gerhard Wingender,
- Ihsan Gursel,
- Mehmet Ozturk,
- Mehmet Inan
Affiliations
- Sibel Kalyoncu
- Izmir Biomedicine and Genome Center
- Semiramis Yilmaz
- Izmir Biomedicine and Genome Center
- Ayca Zeybek Kuyucu
- Izmir Biomedicine and Genome Center
- Dogu Sayili
- Izmir Biomedicine and Genome Center
- Olcay Mert
- Izmir Biomedicine and Genome Center
- Hakan Soyturk
- Izmir Biomedicine and Genome Center
- Seyda Gullu
- Izmir Biomedicine and Genome Center
- Huseyin Akinturk
- Izmir Biomedicine and Genome Center
- Erhan Citak
- Izmir Biomedicine and Genome Center
- Merve Arslan
- Izmir Biomedicine and Genome Center
- Melda Guray Taskinarda
- Izmir Biomedicine and Genome Center
- Ibrahim Oguzhan Tarman
- Izmir Biomedicine and Genome Center
- Gizem Yilmazer Altun
- Izmir Biomedicine and Genome Center
- Ceren Ozer
- Izmir Biomedicine and Genome Center
- Ridvan Orkut
- Izmir Biomedicine and Genome Center
- Aysegul Demirtas
- Izmir Biomedicine and Genome Center
- Idil Tilmensagir
- Izmir Biomedicine and Genome Center
- Umur Keles
- Izmir Biomedicine and Genome Center
- Ceren Ulker
- Izmir Biomedicine and Genome Center
- Gizem Aralan
- Izmir Biomedicine and Genome Center
- Yavuz Mercan
- Izmir Biomedicine and Genome Center
- Muge Ozkan
- Izmir Biomedicine and Genome Center
- Hasan Onur Caglar
- Izmir Biomedicine and Genome Center
- Gizem Arik
- Izmir Biomedicine and Genome Center
- Mehmet Can Ucar
- Izmir Biomedicine and Genome Center
- Muzaffer Yildirim
- Izmir Biomedicine and Genome Center
- Tugce Canavar Yildirim
- Izmir Biomedicine and Genome Center
- Dilara Karadag
- Izmir Biomedicine and Genome Center
- Erhan Bal
- Izmir Biomedicine and Genome Center
- Aybike Erdogan
- Izmir Biomedicine and Genome Center
- Serif Senturk
- Izmir Biomedicine and Genome Center
- Serdar Uzar
- Pendik Veterinary Research and Control Institute
- Hakan Enul
- Pendik Veterinary Research and Control Institute
- Cumhur Adiay
- Pendik Veterinary Research and Control Institute
- Fahriye Sarac
- Pendik Veterinary Research and Control Institute
- Arzu Tas Ekiz
- Marmara Research Center, TUBITAK
- Irem Abaci
- Marmara Research Center, TUBITAK
- Ozge Aksoy
- Marmara Research Center, TUBITAK
- Hivda Ulbegi Polat
- Marmara Research Center, TUBITAK
- Saban Tekin
- Marmara Research Center, TUBITAK
- Stefan Dimitrov
- Izmir Biomedicine and Genome Center
- Aykut Ozkul
- Ankara University
- Gerhard Wingender
- Izmir Biomedicine and Genome Center
- Ihsan Gursel
- Izmir Biomedicine and Genome Center
- Mehmet Ozturk
- Izmir Biomedicine and Genome Center
- Mehmet Inan
- Izmir Biomedicine and Genome Center
- DOI
- https://doi.org/10.1038/s41598-023-32021-9
- Journal volume & issue
-
Vol. 13,
no. 1
pp. 1 – 14
Abstract
Abstract Recombinant protein-based SARS-CoV-2 vaccines are needed to fill the vaccine equity gap. Because protein-subunit based vaccines are easier and cheaper to produce and do not require special storage/transportation conditions, they are suitable for low-/middle-income countries. Here, we report our vaccine development studies with the receptor binding domain of the SARS-CoV-2 Delta Plus strain (RBD-DP) which caused increased hospitalizations compared to other variants. First, we expressed RBD-DP in the Pichia pastoris yeast system and upscaled it to a 5-L fermenter for production. After three-step purification, we obtained RBD-DP with > 95% purity from a protein yield of > 1 g/L of supernatant. Several biophysical and biochemical characterizations were performed to confirm its identity, stability, and functionality. Then, it was formulated in different contents with Alum and CpG for mice immunization. After three doses of immunization, IgG titers from sera reached to > 106 and most importantly it showed high T-cell responses which are required for an effective vaccine to prevent severe COVID-19 disease. A live neutralization test was performed with both the Wuhan strain (B.1.1.7) and Delta strain (B.1.617.2) and it showed high neutralization antibody content for both strains. A challenge study with SARS-CoV-2 infected K18-hACE2 transgenic mice showed good immunoprotective activity with no viruses in the lungs and no lung inflammation for all immunized mice.
