Journal Article Research Support, Non-U.S. Gov't Epublish 12 citations
PubMed: 36997624 DOI: 10.1038/s41598-023-32021-9
Process development for an effective COVID-19 vaccine candidate harboring recombinant SARS-CoV-2 delta plus receptor binding domain produced by Pichia pastoris.
Sci Rep 13(1):5224
Authors: 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
- Izmir Biomedicine and Genome Center, Izmir, Turkey.
- VIB-UGent Center for Medical Biotechnology, Gent, Belgium.
- Izmir International Biomedicine and Genome Institute, Dokuz Eylul University, Izmir, Turkey.
- Lund University, Lund, Sweden.
- Erzurum Technical University, Erzurum, Turkey.
- Ankara Medipol University, Ankara, Turkey.
- Imperial College London, London, UK.
- Izmir Tinaztepe University, Izmir, Turkey.
- Pendik Veterinary Research and Control Institute, Istanbul, Turkey.
- Marmara Research Center, TUBITAK, Kocaeli, Turkey.
- University of Health Sciences, Istanbul, Turkey.
- Ankara University, Ankara, Turkey.
- Izmir Biomedicine and Genome Center, Izmir, Turkey.
- Akdeniz University, Antalya, Turkey.
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 > 10 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.
MeSH
Natural Science Disciplines, Immunization, T-Lymphocytes, Lung, Sars Virus, Pichia, Sars-Cov-2, Covid-19 Vaccines