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 G. 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; Jason J. Lavinder; Gregory C. Ippolito; Jennifer A. Maynard; Ilya J. Finkelstein; Jason S. McLellan

doi:10.1126/SCIENCE.ABD0826

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 G. 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, Jason J. LavinderGregory C. Ippolito, Jennifer A. Maynard, Ilya J. Finkelstein, Jason S. McLellan

Research output: Contribution to journal › Article › peer-review

716 Scopus citations

Abstract

The coronavirus disease 2019 (COVID-19) pandemic has led to accelerated efforts to develop therapeutics and vaccines. A key target of these efforts is the spike (S) protein, which is metastable and difficult to produce recombinantly. We characterized 100 structure-guided spike designs and identified 26 individual substitutions that increased protein yields and stability. Testing combinations of beneficial substitutions resulted in the identification of HexaPro, a variant with six beneficial proline substitutions exhibiting higher expression than its parental construct (by a factor of 10) as well as the ability to withstand heat stress, storage at room temperature, and three freeze-thaw cycles. A cryo-electron microscopy structure of HexaPro at a resolution of 3.2 angstroms 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 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

Original language	English (US)
Pages (from-to)	1501-1505
Number of pages	5
Journal	Science
Volume	369
Issue number	6510
DOIs	https://doi.org/10.1126/SCIENCE.ABD0826
State	Published - Sep 18 2020

Keywords

Amino Acid Substitution
Betacoronavirus/chemistry
COVID-19 Vaccines
Coronavirus Infections/prevention & control
Cryoelectron Microscopy
Humans
Proline/chemistry
Protein Domains
Protein Stability
SARS-CoV-2
Spike Glycoprotein, Coronavirus/chemistry
Viral Vaccines/chemistry

ASJC Scopus subject areas

General

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10.1126/SCIENCE.ABD0826

Cite this

Hsieh, C. L., Goldsmith, J. A., Schaub, J. M., DiVenere, A. M., Kuo, H. C., Javanmardi, K., Le, K. C., Wrapp, D., Lee, A. G., Liu, Y., Chou, C. W., Byrne, P. O., Hjorth, C. K., Johnson, N. V., Ludes-Meyers, J., Nguyen, A. W., Park, J., Wang, N., Amengor, D., ... McLellan, J. S. (2020). Structure-based design of prefusion-stabilized SARS-CoV-2 spikes. Science, 369(6510), 1501-1505. https://doi.org/10.1126/SCIENCE.ABD0826

Hsieh, CL, Goldsmith, JA, Schaub, JM, DiVenere, AM, Kuo, HC, Javanmardi, K, Le, KC, Wrapp, D, Lee, AG, Liu, Y, Chou, CW, Byrne, PO, Hjorth, CK, Johnson, NV, Ludes-Meyers, J, Nguyen, AW, Park, J, Wang, N, Amengor, D, Lavinder, JJ, Ippolito, GC, Maynard, JA, Finkelstein, IJ & McLellan, JS 2020, 'Structure-based design of prefusion-stabilized SARS-CoV-2 spikes', Science, vol. 369, no. 6510, pp. 1501-1505. https://doi.org/10.1126/SCIENCE.ABD0826

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title = "Structure-based design of prefusion-stabilized SARS-CoV-2 spikes",

abstract = "The coronavirus disease 2019 (COVID-19) pandemic has led to accelerated efforts to develop therapeutics and vaccines. A key target of these efforts is the spike (S) protein, which is metastable and difficult to produce recombinantly. We characterized 100 structure-guided spike designs and identified 26 individual substitutions that increased protein yields and stability. Testing combinations of beneficial substitutions resulted in the identification of HexaPro, a variant with six beneficial proline substitutions exhibiting higher expression than its parental construct (by a factor of 10) as well as the ability to withstand heat stress, storage at room temperature, and three freeze-thaw cycles. A cryo-electron microscopy structure of HexaPro at a resolution of 3.2 angstroms 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 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).",

keywords = "Amino Acid Substitution, Betacoronavirus/chemistry, COVID-19 Vaccines, Coronavirus Infections/prevention & control, Cryoelectron Microscopy, Humans, Proline/chemistry, Protein Domains, Protein Stability, SARS-CoV-2, Spike Glycoprotein, Coronavirus/chemistry, Viral Vaccines/chemistry",

author = "Hsieh, {Ching Lin} and Goldsmith, {Jory A.} and Schaub, {Jeffrey M.} and DiVenere, {Andrea M.} and Kuo, {Hung Che} and Kamyab Javanmardi and Le, {Kevin C.} and Daniel Wrapp and Lee, {Alison G.} and Yutong Liu and Chou, {Chia Wei} and Byrne, {Patrick O.} and Hjorth, {Christy K.} and Johnson, {Nicole V.} and John Ludes-Meyers and Nguyen, {Annalee W.} and Juyeon Park and Nianshuang Wang and Dzifa Amengor and Lavinder, {Jason J.} and Ippolito, {Gregory C.} and Maynard, {Jennifer A.} and Finkelstein, {Ilya J.} and McLellan, {Jason S.}",

note = "Copyright {\textcopyright} 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.",

year = "2020",

month = sep,

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doi = "10.1126/SCIENCE.ABD0826",

language = "English (US)",

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AU - Hsieh, Ching Lin

AU - Goldsmith, Jory A.

AU - Schaub, Jeffrey M.

AU - DiVenere, Andrea M.

AU - Kuo, Hung Che

AU - Javanmardi, Kamyab

AU - Le, Kevin C.

AU - Wrapp, Daniel

AU - Lee, Alison G.

AU - Liu, Yutong

AU - Chou, Chia Wei

AU - Byrne, Patrick O.

AU - Hjorth, Christy K.

AU - Johnson, Nicole V.

AU - Ludes-Meyers, John

AU - Nguyen, Annalee W.

AU - Park, Juyeon

AU - Wang, Nianshuang

AU - Amengor, Dzifa

AU - Lavinder, Jason J.

AU - Ippolito, Gregory C.

AU - Maynard, Jennifer A.

AU - Finkelstein, Ilya J.

AU - McLellan, Jason S.

PY - 2020/9/18

Y1 - 2020/9/18

N2 - The coronavirus disease 2019 (COVID-19) pandemic has led to accelerated efforts to develop therapeutics and vaccines. A key target of these efforts is the spike (S) protein, which is metastable and difficult to produce recombinantly. We characterized 100 structure-guided spike designs and identified 26 individual substitutions that increased protein yields and stability. Testing combinations of beneficial substitutions resulted in the identification of HexaPro, a variant with six beneficial proline substitutions exhibiting higher expression than its parental construct (by a factor of 10) as well as the ability to withstand heat stress, storage at room temperature, and three freeze-thaw cycles. A cryo-electron microscopy structure of HexaPro at a resolution of 3.2 angstroms 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 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

AB - The coronavirus disease 2019 (COVID-19) pandemic has led to accelerated efforts to develop therapeutics and vaccines. A key target of these efforts is the spike (S) protein, which is metastable and difficult to produce recombinantly. We characterized 100 structure-guided spike designs and identified 26 individual substitutions that increased protein yields and stability. Testing combinations of beneficial substitutions resulted in the identification of HexaPro, a variant with six beneficial proline substitutions exhibiting higher expression than its parental construct (by a factor of 10) as well as the ability to withstand heat stress, storage at room temperature, and three freeze-thaw cycles. A cryo-electron microscopy structure of HexaPro at a resolution of 3.2 angstroms 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 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

KW - Amino Acid Substitution

KW - Betacoronavirus/chemistry

KW - COVID-19 Vaccines

KW - Coronavirus Infections/prevention & control

KW - Cryoelectron Microscopy

KW - Humans

KW - Proline/chemistry

KW - Protein Domains

KW - Protein Stability

KW - SARS-CoV-2

KW - Spike Glycoprotein, Coronavirus/chemistry

KW - Viral Vaccines/chemistry

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DO - 10.1126/SCIENCE.ABD0826

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SN - 0036-8075

VL - 369

SP - 1501

EP - 1505

JO - Science

JF - Science

IS - 6510

ER -

Structure-based design of prefusion-stabilized SARS-CoV-2 spikes

Abstract

Keywords

ASJC Scopus subject areas

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