Process development for an effective COVID-19 vaccine candidate harboring recombinant SARS-CoV-2 delta plus receptor binding domain produced by Pichia pastoris
(2023) In Scientific Reports 13(1).- 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... (More)
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.
(Less)
- author
- organization
- publishing date
- 2023
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Scientific Reports
- volume
- 13
- issue
- 1
- article number
- 5224
- publisher
- Nature Publishing Group
- external identifiers
-
- pmid:36997624
- scopus:85151316385
- ISSN
- 2045-2322
- DOI
- 10.1038/s41598-023-32021-9
- language
- English
- LU publication?
- yes
- id
- e5df48d7-6f50-48fa-adb4-79894b44a5d6
- date added to LUP
- 2023-05-22 15:15:20
- date last changed
- 2024-04-19 22:01:21
@article{e5df48d7-6f50-48fa-adb4-79894b44a5d6,
abstract = {{<p>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<sup>6</sup> 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.</p>}},
author = {{Kalyoncu, Sibel and Yilmaz, Semiramis and Kuyucu, Ayca Zeybek and Sayili, Dogu and Mert, Olcay and Soyturk, Hakan and Gullu, Seyda and Akinturk, Huseyin and Citak, Erhan and Arslan, Merve and Taskinarda, Melda Guray and Tarman, Ibrahim Oguzhan and Altun, Gizem Yilmazer and Ozer, Ceren and Orkut, Ridvan and Demirtas, Aysegul and Tilmensagir, Idil and Keles, Umur and Ulker, Ceren and Aralan, Gizem and Mercan, Yavuz and Ozkan, Muge and Caglar, Hasan Onur and Arik, Gizem and Ucar, Mehmet Can and Yildirim, Muzaffer and Yildirim, Tugce Canavar and Karadag, Dilara and Bal, Erhan and Erdogan, Aybike and Senturk, Serif and Uzar, Serdar and Enul, Hakan and Adiay, Cumhur and Sarac, Fahriye and Ekiz, Arzu Tas and Abaci, Irem and Aksoy, Ozge and Polat, Hivda Ulbegi and Tekin, Saban and Dimitrov, Stefan and Ozkul, Aykut and Wingender, Gerhard and Gursel, Ihsan and Ozturk, Mehmet and Inan, Mehmet}},
issn = {{2045-2322}},
language = {{eng}},
number = {{1}},
publisher = {{Nature Publishing Group}},
series = {{Scientific Reports}},
title = {{Process development for an effective COVID-19 vaccine candidate harboring recombinant SARS-CoV-2 delta plus receptor binding domain produced by Pichia pastoris}},
url = {{http://dx.doi.org/10.1038/s41598-023-32021-9}},
doi = {{10.1038/s41598-023-32021-9}},
volume = {{13}},
year = {{2023}},
}