Trimeric SARS-CoV-2 Spike Proteins Produced from CHO Cells in Bioreactors Are High-Quality Antigens

Details

Ressource 1Download: Trimeric SARS-CoV-2 Spike Proteins Produced from CHO Cells in Bioreactors Are High-Quality Antigens.pdf (3329.10 [Ko])
State: Public
Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_E313FE209395
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Trimeric SARS-CoV-2 Spike Proteins Produced from CHO Cells in Bioreactors Are High-Quality Antigens
Journal
Processes
Author(s)
Pino Paco, Kint Joeri, Kiseljak Divor, Agnolon Valentina, Corradin Giampietro, Kajava Andrey V., Rovero Paolo, Dijkman Ronald, den Hartog Gerco, McLellan Jason S., Byrne Patrick O., Wurm Maria J., Wurm Florian M.
ISSN
2227-9717
Publication state
Published
Issued date
25/11/2020
Volume
8
Number
12
Pages
1539
Language
english
Abstract
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
Process Chemistry and Technology, Chemical Engineering (miscellaneous), Bioengineering
Web of science
Open Access
Yes
Create date
13/01/2021 9:26
Last modification date
16/04/2024 6:24
Usage data