Structure-based Design of Prefusion-stabilized SARS-CoV-2 Spikes
Jory A Goldsmith,
Jeffrey M. Schaub,
Andrea M DiVenere,
Kevin C Le,
Alison Gene-Wei Lee,
Patrick O. Byrne,
Christy K Hjorth,
Nicole V Johnson,
Annalee W Nguyen,
Jennifer A Maynard,
Ilya J Finkelstein,
Jason S McLellan
Posted 30 May 2020
bioRxiv DOI: 10.1101/2020.05.30.125484 (published DOI: 10.1126/science.abd0826)
Posted 30 May 2020
The COVID-19 pandemic caused by the novel coronavirus SARS-CoV-2 has led to accelerated efforts to develop therapeutics, diagnostics, and vaccines to mitigate this public health emergency. A key target of these efforts is the spike (S) protein, a large trimeric class I fusion protein that is metastable and difficult to produce recombinantly in large quantities. Here, we designed and expressed over 100 structure-guided spike variants based upon a previously determined cryo-EM structure of the prefusion SARS-CoV-2 spike. Biochemical, biophysical and structural characterization of these variants identified numerous individual substitutions that increased protein yields and stability. The best variant, HexaPro, has six beneficial proline substitutions leading to ~10-fold higher expression than its parental construct and is able to withstand heat stress, storage at room temperature, and multiple freeze-thaws. A 3.2 Å-resolution cryo-EM structure of HexaPro confirmed that it retains the prefusion spike conformation. High-yield production of a stabilized prefusion spike protein will accelerate the development of vaccines and serological diagnostics for SARS-CoV-2. ### Competing Interest Statement N.W. and J.S.M. are inventors on U.S. patent application no. 62/412,703 (Prefusion Coronavirus Spike Proteins and Their Use). D.W., N.W. and J.S.M. are inventors on U.S. patent application no. 62/972,886 (2019-nCoV Vaccine). C.-L.H., J.A.G., J.M.S., C.-W.C., A.M.D., K.J., H.-C.K., D.W., P.O.B., C.K.H., N.V.J., N.W., J.A.M., I.J.F., and J.S.M. are inventors on U.S. patent application no. 63/032,502 (Engineered Coronavirus Spike (S) Protein and Methods of Use Thereof).
- Downloaded 4,243 times
- Download rankings, all-time:
- Site-wide: 2,488
- In microbiology: 181
- Year to date:
- Site-wide: 2,941
- Since beginning of last month:
- Site-wide: 1,944
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!