LAPSE:2021.0541
Published Article
LAPSE:2021.0541
Trimeric SARS-CoV-2 Spike Proteins Produced from CHO Cells in Bioreactors Are High-Quality Antigens
Paco Pino, Joeri Kint, Divor Kiseljak, Valentina Agnolon, Giampietro Corradin, Andrey V. Kajava, Paolo Rovero, Ronald Dijkman, Gerco den Hartog, Jason S. McLellan, Patrick O. Byrne, Maria J. Wurm, Florian M. Wurm
June 21, 2021
The spike protein of the pandemic human corona virus is essential for its entry into human cells. In fact, most neutralizing antibodies against Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2) are directed against the Virus-surface exposed spike protein, making it the antigen of choice for use in vaccines and diagnostic tests. In the current pandemic context, global demand for spike proteins has rapidly increased and could exceed hundreds of grams to kilograms annually. Coronavirus spikes are large heavily glycosylated homo-trimeric complexes, with inherent instability. The poor manufacturability now threatens the availability of these proteins for vaccines and diagnostic tests. Here, we outline scalable, Good Manufacturing Practice (GMP) compliant, and chemically defined processes for the production of two cell-secreted stabilized forms of the trimeric spike proteins (Wuhan and D614G variant). The processes are chemically defined and based on clonal suspension-CHO cell populations and on protein purification via a two-step scalable downstream process. The trimeric conformation was confirmed using electron microscopy and HPLC analysis. Binding to susceptible cells was shown using a virus-inhibition assay. The diagnostic sensitivity and specificity for detection of serum SARS-CoV-2-specific-immunoglobulin molecules was found to exceed that of spike fragments (Spike subunit-1, S1 and Receptor Binding Domain, RBD). The process described here will enable production of sufficient high-quality trimeric spike protein to meet the global demand for SARS-CoV-2 diagnostic tests and potentially vaccines.
Keywords
CHO cells, diagnostics, manufacturability, SARS-CoV-2, trimeric spike, vaccines
Subject
Suggested Citation
Pino P, Kint J, Kiseljak D, Agnolon V, Corradin G, Kajava AV, Rovero P, Dijkman R, den Hartog G, McLellan JS, Byrne PO, Wurm MJ, Wurm FM. Trimeric SARS-CoV-2 Spike Proteins Produced from CHO Cells in Bioreactors Are High-Quality Antigens. (2021). LAPSE:2021.0541
Author Affiliations
Pino P: ExcellGene SA, CH1870 Monthey, Switzerland
Kint J: ExcellGene SA, CH1870 Monthey, Switzerland
Kiseljak D: ExcellGene SA, CH1870 Monthey, Switzerland
Agnolon V: Department of Immunology and Allergy, Centre Hospitalier Universitaire Vaudois, (CHUV), 1011 Lausanne, Switzerland
Corradin G: Department of Immunology and Allergy, Centre Hospitalier Universitaire Vaudois, (CHUV), 1011 Lausanne, Switzerland
Kajava AV: Centre de Recherche en Biologie cellulaire de Montpellier, Université Montpellier, 34293 Montpellier, France [ORCID]
Rovero P: Unit Peptide and Protein Chemistry and Biology, Department of NeuroFarBra, University of Florence, 50019 Sesto Florentino, Italy [ORCID]
Dijkman R: Institute for Infectious Diseases, University of Bern, 3001 Bern, Switzerland; Institute of Virology and Immunology (IVI), 3012 Bern, Switzerland; Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, 3012 Bern, Switze [ORCID]
den Hartog G: Department of Immuno-Surveillance, Centre for Immunology of Infectious Diseases and Vaccines, National Institute for Public Health and the Environment, 3741MA Bilthoven, The Netherlands [ORCID]
McLellan JS: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712, USA
Byrne PO: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712, USA [ORCID]
Wurm MJ: ExcellGene SA, CH1870 Monthey, Switzerland
Wurm FM: ExcellGene SA, CH1870 Monthey, Switzerland; Life Science Faculty, Swiss Federal Institute of Technology Lausanne (EPFL), 1015 Lausanne, Switzerland
Journal Name
Processes
Volume
8
Issue
12
Article Number
E1539
Year
2020
Publication Date
2020-11-25
Published Version
ISSN
2227-9717
Version Comments
Original Submission
Other Meta
PII: pr8121539, Publication Type: Journal Article
Record Map
Published Article

LAPSE:2021.0541
This Record
External Link

doi:10.3390/pr8121539
Publisher Version
Download
Files
[Download 1v1.pdf] (3.3 MB)
Jun 21, 2021
Main Article
License
CC BY 4.0
Meta
Record Statistics
Record Views
297
Version History
[v1] (Original Submission)
Jun 21, 2021
 
Verified by curator on
Jun 21, 2021
This Version Number
v1
Citations
Most Recent
This Version
URL Here
https://psecommunity.org/LAPSE:2021.0541
 
Original Submitter
Calvin Tsay
Links to Related Works
Directly Related to This Work
Publisher Version