An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike

Michael Schoof; Bryan Faust; Reuben A. Saunders; Smriti Sangwan; V. Rezelj; Nick Hoppe; Morgane Boone; Christian B. Billesbølle; Cristina Puchades; Caleigh M. Azumaya; Huong T. Kratochvil; Marcell Zimanyi; Ishan Deshpande; Jiahao Liang; Sasha Dickinson; Henry C. Nguyen; Cynthia M. Chio; Gregory E. Merz; Michael C. Thompson; Devan Diwanji; Kaitlin Schaefer; Aditya A. Anand; Niv Dobzinski; Beth Shoshana Zha; Camille R. Simoneau; Kristoffer Leon; Kris M. White1; Un Seng Chio; Meghna Gupta; Mingliang Jin; Fei Li; Yanxin Liu; Kaihua Zhang; David Bulkley; Ming Sun; Amber M. Smith; Alexandrea N. Rizo; Frank Moss; Axel F. Brilot; Sergei Pourmal; Raphael Trenker; Thomas Pospiech; Sayan Gupta; Benjamin Barsi-Rhyne; Vladislav Belyy; Andrew W. Barile-Hill1; Silke Nock; Yuwei Liu; Nevan J. Krogan; Corie Y. Ralston; Danielle L. Swaney; Adolfo Garciá-Sastre; Melanie Ott; Marco Vignuzzi; Peter Walter; Aashish Manglik

doi:10.1126/science.abe3255

An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike

Michael Schoof^*, Bryan Faust, Reuben A. Saunders, Smriti Sangwan, V. Rezelj, Nick Hoppe, Morgane Boone, Christian B. Billesbølle, Cristina Puchades, Caleigh M. Azumaya, Huong T. Kratochvil, Marcell Zimanyi, Ishan Deshpande, Jiahao Liang, Sasha Dickinson, Henry C. Nguyen, Cynthia M. Chio, Gregory E. Merz, Michael C. Thompson, Devan DiwanjiKaitlin Schaefer, Aditya A. Anand, Niv Dobzinski, Beth Shoshana Zha, Camille R. Simoneau, Kristoffer Leon, Kris M. White1, Un Seng Chio, Meghna Gupta, Mingliang Jin, Fei Li, Yanxin Liu, Kaihua Zhang, David Bulkley, Ming Sun, Amber M. Smith, Alexandrea N. Rizo, Frank Moss, Axel F. Brilot, Sergei Pourmal, Raphael Trenker, Thomas Pospiech, Sayan Gupta, Benjamin Barsi-Rhyne, Vladislav Belyy, Andrew W. Barile-Hill1, Silke Nock, Yuwei Liu, Nevan J. Krogan, Corie Y. Ralston, Danielle L. Swaney, Adolfo Garciá-Sastre, Melanie Ott, Marco Vignuzzi, Peter Walter^*, Aashish Manglik^*

^*Corresponding author for this work

Pharmacology

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

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Abstract

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus enters host cells via an interaction between its Spike protein and the host cell receptor angiotensin-converting enzyme 2 (ACE2). By screening a yeast surface-displayed library of synthetic nanobody sequences, we developed nanobodies that disrupt the interaction between Spike and ACE2. Cryo-electron microscopy (cryo-EM) revealed that one nanobody, Nb6, binds Spike in a fully inactive conformation with its receptor binding domains locked into their inaccessible down state, incapable of binding ACE2. Affinity maturation and structure-guided design of multivalency yielded a trivalent nanobody, mNb6-tri, with femtomolar affinity for Spike and picomolar neutralization of SARS-CoV-2 infection. mNb6-tri retains function after aerosolization, lyophilization, and heat treatment, which enables aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia.

Original language	English (US)
Pages (from-to)	1473-1479
Number of pages	7
Journal	Science
Volume	370
Issue number	6523
DOIs	https://doi.org/10.1126/science.abe3255
State	Published - Dec 18 2020

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Schoof, M., Faust, B., Saunders, R. A., Sangwan, S., Rezelj, V., Hoppe, N., Boone, M., Billesbølle, C. B., Puchades, C., Azumaya, C. M., Kratochvil, H. T., Zimanyi, M., Deshpande, I., Liang, J., Dickinson, S., Nguyen, H. C., Chio, C. M., Merz, G. E., Thompson, M. C., ... Manglik, A. (2020). An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike. Science, 370(6523), 1473-1479. https://doi.org/10.1126/science.abe3255

@article{5138ef4beba443ac9865b21b8e8da4a8,

title = "An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike",

abstract = "The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus enters host cells via an interaction between its Spike protein and the host cell receptor angiotensin-converting enzyme 2 (ACE2). By screening a yeast surface-displayed library of synthetic nanobody sequences, we developed nanobodies that disrupt the interaction between Spike and ACE2. Cryo-electron microscopy (cryo-EM) revealed that one nanobody, Nb6, binds Spike in a fully inactive conformation with its receptor binding domains locked into their inaccessible down state, incapable of binding ACE2. Affinity maturation and structure-guided design of multivalency yielded a trivalent nanobody, mNb6-tri, with femtomolar affinity for Spike and picomolar neutralization of SARS-CoV-2 infection. mNb6-tri retains function after aerosolization, lyophilization, and heat treatment, which enables aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia.",

author = "Michael Schoof and Bryan Faust and Saunders, {Reuben A.} and Smriti Sangwan and V. Rezelj and Nick Hoppe and Morgane Boone and Billesb{\o}lle, {Christian B.} and Cristina Puchades and Azumaya, {Caleigh M.} and Kratochvil, {Huong T.} and Marcell Zimanyi and Ishan Deshpande and Jiahao Liang and Sasha Dickinson and Nguyen, {Henry C.} and Chio, {Cynthia M.} and Merz, {Gregory E.} and Thompson, {Michael C.} and Devan Diwanji and Kaitlin Schaefer and Anand, {Aditya A.} and Niv Dobzinski and Zha, {Beth Shoshana} and Simoneau, {Camille R.} and Kristoffer Leon and White1, {Kris M.} and Chio, {Un Seng} and Meghna Gupta and Mingliang Jin and Fei Li and Yanxin Liu and Kaihua Zhang and David Bulkley and Ming Sun and Smith, {Amber M.} and Rizo, {Alexandrea N.} and Frank Moss and Brilot, {Axel F.} and Sergei Pourmal and Raphael Trenker and Thomas Pospiech and Sayan Gupta and Benjamin Barsi-Rhyne and Vladislav Belyy and Barile-Hill1, {Andrew W.} and Silke Nock and Yuwei Liu and Krogan, {Nevan J.} and Ralston, {Corie Y.} and Swaney, {Danielle L.} and Adolfo Garci{\'a}-Sastre and Melanie Ott and Marco Vignuzzi and Peter Walter and Aashish Manglik",

year = "2020",

month = dec,

day = "18",

doi = "10.1126/science.abe3255",

language = "English (US)",

volume = "370",

pages = "1473--1479",

journal = "Science",

issn = "0036-8075",

publisher = "American Association for the Advancement of Science",

number = "6523",

}

Schoof, M, Faust, B, Saunders, RA, Sangwan, S, Rezelj, V, Hoppe, N, Boone, M, Billesbølle, CB, Puchades, C, Azumaya, CM, Kratochvil, HT, Zimanyi, M, Deshpande, I, Liang, J, Dickinson, S, Nguyen, HC, Chio, CM, Merz, GE, Thompson, MC, Diwanji, D, Schaefer, K, Anand, AA, Dobzinski, N, Zha, BS, Simoneau, CR, Leon, K, White1, KM, Chio, US, Gupta, M, Jin, M, Li, F, Liu, Y, Zhang, K, Bulkley, D, Sun, M, Smith, AM, Rizo, AN, Moss, F, Brilot, AF, Pourmal, S, Trenker, R, Pospiech, T, Gupta, S, Barsi-Rhyne, B, Belyy, V, Barile-Hill1, AW, Nock, S, Liu, Y, Krogan, NJ, Ralston, CY, Swaney, DL, Garciá-Sastre, A, Ott, M, Vignuzzi, M, Walter, P & Manglik, A 2020, 'An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike', Science, vol. 370, no. 6523, pp. 1473-1479. https://doi.org/10.1126/science.abe3255

TY - JOUR

T1 - An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike

AU - Schoof, Michael

AU - Faust, Bryan

AU - Saunders, Reuben A.

AU - Sangwan, Smriti

AU - Rezelj, V.

AU - Hoppe, Nick

AU - Boone, Morgane

AU - Billesbølle, Christian B.

AU - Puchades, Cristina

AU - Azumaya, Caleigh M.

AU - Kratochvil, Huong T.

AU - Zimanyi, Marcell

AU - Deshpande, Ishan

AU - Liang, Jiahao

AU - Dickinson, Sasha

AU - Nguyen, Henry C.

AU - Chio, Cynthia M.

AU - Merz, Gregory E.

AU - Thompson, Michael C.

AU - Diwanji, Devan

AU - Schaefer, Kaitlin

AU - Anand, Aditya A.

AU - Dobzinski, Niv

AU - Zha, Beth Shoshana

AU - Simoneau, Camille R.

AU - Leon, Kristoffer

AU - White1, Kris M.

AU - Chio, Un Seng

AU - Gupta, Meghna

AU - Jin, Mingliang

AU - Li, Fei

AU - Liu, Yanxin

AU - Zhang, Kaihua

AU - Bulkley, David

AU - Sun, Ming

AU - Smith, Amber M.

AU - Rizo, Alexandrea N.

AU - Moss, Frank

AU - Brilot, Axel F.

AU - Pourmal, Sergei

AU - Trenker, Raphael

AU - Pospiech, Thomas

AU - Gupta, Sayan

AU - Barsi-Rhyne, Benjamin

AU - Belyy, Vladislav

AU - Barile-Hill1, Andrew W.

AU - Nock, Silke

AU - Liu, Yuwei

AU - Krogan, Nevan J.

AU - Ralston, Corie Y.

AU - Swaney, Danielle L.

AU - Garciá-Sastre, Adolfo

AU - Ott, Melanie

AU - Vignuzzi, Marco

AU - Walter, Peter

AU - Manglik, Aashish

PY - 2020/12/18

Y1 - 2020/12/18

N2 - The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus enters host cells via an interaction between its Spike protein and the host cell receptor angiotensin-converting enzyme 2 (ACE2). By screening a yeast surface-displayed library of synthetic nanobody sequences, we developed nanobodies that disrupt the interaction between Spike and ACE2. Cryo-electron microscopy (cryo-EM) revealed that one nanobody, Nb6, binds Spike in a fully inactive conformation with its receptor binding domains locked into their inaccessible down state, incapable of binding ACE2. Affinity maturation and structure-guided design of multivalency yielded a trivalent nanobody, mNb6-tri, with femtomolar affinity for Spike and picomolar neutralization of SARS-CoV-2 infection. mNb6-tri retains function after aerosolization, lyophilization, and heat treatment, which enables aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia.

AB - The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus enters host cells via an interaction between its Spike protein and the host cell receptor angiotensin-converting enzyme 2 (ACE2). By screening a yeast surface-displayed library of synthetic nanobody sequences, we developed nanobodies that disrupt the interaction between Spike and ACE2. Cryo-electron microscopy (cryo-EM) revealed that one nanobody, Nb6, binds Spike in a fully inactive conformation with its receptor binding domains locked into their inaccessible down state, incapable of binding ACE2. Affinity maturation and structure-guided design of multivalency yielded a trivalent nanobody, mNb6-tri, with femtomolar affinity for Spike and picomolar neutralization of SARS-CoV-2 infection. mNb6-tri retains function after aerosolization, lyophilization, and heat treatment, which enables aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia.

UR - http://www.scopus.com/inward/record.url?scp=85097074102&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85097074102&partnerID=8YFLogxK

U2 - 10.1126/science.abe3255

DO - 10.1126/science.abe3255

M3 - Article

C2 - 33154106

AN - SCOPUS:85097074102

SN - 0036-8075

VL - 370

SP - 1473

EP - 1479

JO - Science

JF - Science

IS - 6523

ER -

An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike

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