Trimeric SARS-CoV-2 Spike proteins produced from CHO-cells in bioreactors are high-quality antigens
Andrey V Kajava,
Gerco den Hartog,
Jason S McLellan,
Maria J Wurm,
Florian M Wurm
Posted 16 Nov 2020
bioRxiv DOI: 10.1101/2020.11.15.382044
Posted 16 Nov 2020
The Spike protein of SARS-CoV-2 is essential for virus entry into human cells. In fact, most neutralizing antibodies against SARS-CoV-2 are directed against Spike, 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. Their poor manufacturability now threatens the availability of these proteins for vaccines and diagnostic tests. Here, we outline a scalable, GMP-compliant, chemically defined process for the production of a cell-secreted, stabilized form of the trimeric Spike protein. The process is chemically defined and based on clonal, suspension-CHO cell populations and on protein purification via a two-step, scalable down-stream 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 the detection of serum SARS-CoV-2 specific IgG1 was investigated and found to exceed that of Spike fragments (S1 and RBD). The process described here will enable the production of sufficient, high-quality trimeric Spike protein to meet the global demand for SARS-CoV-2 vaccines and diagnostic tests. ### Competing Interest Statement ExcellGene declares that material resulting from this work is being provided commercially. ExcellGene authors declare that the interpretation of the results is done objectively. Conclusions and interpretation of results of non-ExcellGene authors (EM analysis, inhibition of virus infection, reactivity against patient sera) were solely based on their judgment.
- Downloaded 292 times
- Download rankings, all-time:
- Site-wide: 86,335
- In cell biology: 4,000
- Year to date:
- Site-wide: 34,734
- Since beginning of last month:
- Site-wide: 29,666
Downloads over time
Distribution of downloads per paper, site-wide
- 27 Nov 2020: The website and API now include results pulled from medRxiv as well as bioRxiv.
- 18 Dec 2019: We're pleased to announce PanLingua, a new tool that enables you to search for machine-translated bioRxiv preprints using more than 100 different languages.
- 21 May 2019: PLOS Biology has published a community page about Rxivist.org and its design.
- 10 May 2019: The paper analyzing the Rxivist dataset has been published at eLife.
- 1 Mar 2019: We now have summary statistics about bioRxiv downloads and submissions.
- 8 Feb 2019: Data from Altmetric is now available on the Rxivist details page for every preprint. Look for the "donut" under the download metrics.
- 30 Jan 2019: preLights has featured the Rxivist preprint and written about our findings.
- 22 Jan 2019: Nature just published an article about Rxivist and our data.
- 13 Jan 2019: The Rxivist preprint is live!