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Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate
SARS-CoV-2 protein subunit vaccines are currently being evaluated by multiple manufacturers to address the global vaccine equity gap, and need for low-cost, easy to scale, safe, and effective COVID-19 vaccines. In this paper, we report on the generation of the receptor-binding domain RBD203-N1 yeast...
Autores principales: | , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Authors. Published by Elsevier Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8529586/ https://www.ncbi.nlm.nih.gov/pubmed/34688919 http://dx.doi.org/10.1016/j.pep.2021.106003 |
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author | Chen, Wen-Hsiang Pollet, Jeroen Strych, Ulrich Lee, Jungsoon Liu, Zhuyun Kundu, Rakhi Tyagi Versteeg, Leroy Villar, Maria Jose Adhikari, Rakesh Wei, Junfei Poveda, Cristina Keegan, Brian Bailey, Aaron Oakley Chen, Yi-Lin Gillespie, Portia M. Kimata, Jason T. Zhan, Bin Hotez, Peter J. Bottazzi, Maria Elena |
author_facet | Chen, Wen-Hsiang Pollet, Jeroen Strych, Ulrich Lee, Jungsoon Liu, Zhuyun Kundu, Rakhi Tyagi Versteeg, Leroy Villar, Maria Jose Adhikari, Rakesh Wei, Junfei Poveda, Cristina Keegan, Brian Bailey, Aaron Oakley Chen, Yi-Lin Gillespie, Portia M. Kimata, Jason T. Zhan, Bin Hotez, Peter J. Bottazzi, Maria Elena |
author_sort | Chen, Wen-Hsiang |
collection | PubMed |
description | SARS-CoV-2 protein subunit vaccines are currently being evaluated by multiple manufacturers to address the global vaccine equity gap, and need for low-cost, easy to scale, safe, and effective COVID-19 vaccines. In this paper, we report on the generation of the receptor-binding domain RBD203-N1 yeast expression construct, which produces a recombinant protein capable of eliciting a robust immune response and protection in mice against SARS-CoV-2 challenge infections. The RBD203-N1 antigen was expressed in the yeast Pichia pastoris X33. After fermentation at the 5 L scale, the protein was purified by hydrophobic interaction chromatography followed by anion exchange chromatography. The purified protein was characterized biophysically and biochemically, and after its formulation, the immunogenicity was evaluated in mice. Sera were evaluated for their efficacy using a SARS-CoV-2 pseudovirus assay. The RBD203-N1 protein was expressed with a yield of 492.9 ± 3.0 mg/L of fermentation supernatant. A two-step purification process produced a >96% pure protein with a recovery rate of 55 ± 3% (total yield of purified protein: 270.5 ± 13.2 mg/L fermentation supernatant). The protein was characterized to be a homogeneous monomer that showed a well-defined secondary structure, was thermally stable, antigenic, and when adjuvanted on Alhydrogel in the presence of CpG it was immunogenic and induced high levels of neutralizing antibodies against SARS-CoV-2 pseudovirus. The characteristics of the RBD203-N1 protein-based vaccine show that this candidate is another well suited RBD-based construct for technology transfer to manufacturing entities and feasibility of transition into the clinic to evaluate its immunogenicity and safety in humans. |
format | Online Article Text |
id | pubmed-8529586 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Authors. Published by Elsevier Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-85295862021-10-21 Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate Chen, Wen-Hsiang Pollet, Jeroen Strych, Ulrich Lee, Jungsoon Liu, Zhuyun Kundu, Rakhi Tyagi Versteeg, Leroy Villar, Maria Jose Adhikari, Rakesh Wei, Junfei Poveda, Cristina Keegan, Brian Bailey, Aaron Oakley Chen, Yi-Lin Gillespie, Portia M. Kimata, Jason T. Zhan, Bin Hotez, Peter J. Bottazzi, Maria Elena Protein Expr Purif Article SARS-CoV-2 protein subunit vaccines are currently being evaluated by multiple manufacturers to address the global vaccine equity gap, and need for low-cost, easy to scale, safe, and effective COVID-19 vaccines. In this paper, we report on the generation of the receptor-binding domain RBD203-N1 yeast expression construct, which produces a recombinant protein capable of eliciting a robust immune response and protection in mice against SARS-CoV-2 challenge infections. The RBD203-N1 antigen was expressed in the yeast Pichia pastoris X33. After fermentation at the 5 L scale, the protein was purified by hydrophobic interaction chromatography followed by anion exchange chromatography. The purified protein was characterized biophysically and biochemically, and after its formulation, the immunogenicity was evaluated in mice. Sera were evaluated for their efficacy using a SARS-CoV-2 pseudovirus assay. The RBD203-N1 protein was expressed with a yield of 492.9 ± 3.0 mg/L of fermentation supernatant. A two-step purification process produced a >96% pure protein with a recovery rate of 55 ± 3% (total yield of purified protein: 270.5 ± 13.2 mg/L fermentation supernatant). The protein was characterized to be a homogeneous monomer that showed a well-defined secondary structure, was thermally stable, antigenic, and when adjuvanted on Alhydrogel in the presence of CpG it was immunogenic and induced high levels of neutralizing antibodies against SARS-CoV-2 pseudovirus. The characteristics of the RBD203-N1 protein-based vaccine show that this candidate is another well suited RBD-based construct for technology transfer to manufacturing entities and feasibility of transition into the clinic to evaluate its immunogenicity and safety in humans. The Authors. Published by Elsevier Inc. 2022-02 2021-10-21 /pmc/articles/PMC8529586/ /pubmed/34688919 http://dx.doi.org/10.1016/j.pep.2021.106003 Text en © 2021 The Authors Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Chen, Wen-Hsiang Pollet, Jeroen Strych, Ulrich Lee, Jungsoon Liu, Zhuyun Kundu, Rakhi Tyagi Versteeg, Leroy Villar, Maria Jose Adhikari, Rakesh Wei, Junfei Poveda, Cristina Keegan, Brian Bailey, Aaron Oakley Chen, Yi-Lin Gillespie, Portia M. Kimata, Jason T. Zhan, Bin Hotez, Peter J. Bottazzi, Maria Elena Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate |
title | Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate |
title_full | Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate |
title_fullStr | Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate |
title_full_unstemmed | Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate |
title_short | Yeast-expressed recombinant SARS-CoV-2 receptor binding domain RBD203-N1 as a COVID-19 protein vaccine candidate |
title_sort | yeast-expressed recombinant sars-cov-2 receptor binding domain rbd203-n1 as a covid-19 protein vaccine candidate |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8529586/ https://www.ncbi.nlm.nih.gov/pubmed/34688919 http://dx.doi.org/10.1016/j.pep.2021.106003 |
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