Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Structure-based Design of Prefusion-stabilized SARS-CoV-2 Spikes

Ching-Lin Hsieh, Jory A. Goldsmith, Jeffrey M. Schaub, Andrea M. DiVenere, Hung-Che Kuo, Kamyab Javanmardi, Kevin C. Le, Daniel Wrapp, Alison Gene-Wei Lee, Yutong Liu, Chia-Wei Chou, Patrick O. Byrne, Christy K. Hjorth, Nicole V. Johnson, John Ludes-Meyers, Annalee W. Nguyen, Juyeon Park, Nianshuang Wang, Dzifa Amengor, Jennifer A. Maynard, Ilya J. Finkelstein, View ORCID ProfileJason S. McLellan
doi: https://doi.org/10.1101/2020.05.30.125484
Ching-Lin Hsieh
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jory A. Goldsmith
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jeffrey M. Schaub
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Andrea M. DiVenere
2Department of Chemical Engineering, University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hung-Che Kuo
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kamyab Javanmardi
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Kevin C. Le
2Department of Chemical Engineering, University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Daniel Wrapp
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alison Gene-Wei Lee
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Yutong Liu
2Department of Chemical Engineering, University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Chia-Wei Chou
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Patrick O. Byrne
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Christy K. Hjorth
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nicole V. Johnson
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
John Ludes-Meyers
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Annalee W. Nguyen
2Department of Chemical Engineering, University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Juyeon Park
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nianshuang Wang
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Dzifa Amengor
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jennifer A. Maynard
2Department of Chemical Engineering, University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: maynard@che.utexas.edu ilya@finkelsteinlab.org jmclellan@austin.utexas.edu
Ilya J. Finkelstein
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
3Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: maynard@che.utexas.edu ilya@finkelsteinlab.org jmclellan@austin.utexas.edu
Jason S. McLellan
1Department of Molecular Biosciences, The University of Texas at Austin, Austin, Texas 78712
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Jason S. McLellan
  • For correspondence: maynard@che.utexas.edu ilya@finkelsteinlab.org jmclellan@austin.utexas.edu
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

ABSTRACT

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).

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
Back to top
PreviousNext
Posted May 30, 2020.
Download PDF
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Structure-based Design of Prefusion-stabilized SARS-CoV-2 Spikes
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Structure-based Design of Prefusion-stabilized SARS-CoV-2 Spikes
Ching-Lin Hsieh, Jory A. Goldsmith, Jeffrey M. Schaub, Andrea M. DiVenere, Hung-Che Kuo, Kamyab Javanmardi, Kevin C. Le, Daniel Wrapp, Alison Gene-Wei Lee, Yutong Liu, Chia-Wei Chou, Patrick O. Byrne, Christy K. Hjorth, Nicole V. Johnson, John Ludes-Meyers, Annalee W. Nguyen, Juyeon Park, Nianshuang Wang, Dzifa Amengor, Jennifer A. Maynard, Ilya J. Finkelstein, Jason S. McLellan
bioRxiv 2020.05.30.125484; doi: https://doi.org/10.1101/2020.05.30.125484
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
Structure-based Design of Prefusion-stabilized SARS-CoV-2 Spikes
Ching-Lin Hsieh, Jory A. Goldsmith, Jeffrey M. Schaub, Andrea M. DiVenere, Hung-Che Kuo, Kamyab Javanmardi, Kevin C. Le, Daniel Wrapp, Alison Gene-Wei Lee, Yutong Liu, Chia-Wei Chou, Patrick O. Byrne, Christy K. Hjorth, Nicole V. Johnson, John Ludes-Meyers, Annalee W. Nguyen, Juyeon Park, Nianshuang Wang, Dzifa Amengor, Jennifer A. Maynard, Ilya J. Finkelstein, Jason S. McLellan
bioRxiv 2020.05.30.125484; doi: https://doi.org/10.1101/2020.05.30.125484

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Microbiology
Subject Areas
All Articles
  • Animal Behavior and Cognition (4115)
  • Biochemistry (8818)
  • Bioengineering (6522)
  • Bioinformatics (23466)
  • Biophysics (11792)
  • Cancer Biology (9212)
  • Cell Biology (13326)
  • Clinical Trials (138)
  • Developmental Biology (7439)
  • Ecology (11413)
  • Epidemiology (2066)
  • Evolutionary Biology (15155)
  • Genetics (10439)
  • Genomics (14045)
  • Immunology (9173)
  • Microbiology (22159)
  • Molecular Biology (8814)
  • Neuroscience (47581)
  • Paleontology (350)
  • Pathology (1429)
  • Pharmacology and Toxicology (2492)
  • Physiology (3731)
  • Plant Biology (8082)
  • Scientific Communication and Education (1437)
  • Synthetic Biology (2221)
  • Systems Biology (6039)
  • Zoology (1253)