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Stabilization of the SARS-CoV-2 Spike Receptor-Binding Domain Using Deep Mutational Scanning and Structure-Based Design

The unprecedented global demand for SARS-CoV-2 vaccines has demonstrated the need for highly effective vaccine candidates that are thermostable and amenable to large-scale manufacturing. Nanoparticle immunogens presenting the receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein (S) in repet...

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Autores principales: Ellis, Daniel, Brunette, Natalie, Crawford, Katharine H. D., Walls, Alexandra C., Pham, Minh N., Chen, Chengbo, Herpoldt, Karla-Luise, Fiala, Brooke, Murphy, Michael, Pettie, Deleah, Kraft, John C., Malone, Keara D., Navarro, Mary Jane, Ogohara, Cassandra, Kepl, Elizabeth, Ravichandran, Rashmi, Sydeman, Claire, Ahlrichs, Maggie, Johnson, Max, Blackstone, Alyssa, Carter, Lauren, Starr, Tyler N., Greaney, Allison J., Lee, Kelly K., Veesler, David, Bloom, Jesse D., King, Neil P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8276696/
https://www.ncbi.nlm.nih.gov/pubmed/34267764
http://dx.doi.org/10.3389/fimmu.2021.710263
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author Ellis, Daniel
Brunette, Natalie
Crawford, Katharine H. D.
Walls, Alexandra C.
Pham, Minh N.
Chen, Chengbo
Herpoldt, Karla-Luise
Fiala, Brooke
Murphy, Michael
Pettie, Deleah
Kraft, John C.
Malone, Keara D.
Navarro, Mary Jane
Ogohara, Cassandra
Kepl, Elizabeth
Ravichandran, Rashmi
Sydeman, Claire
Ahlrichs, Maggie
Johnson, Max
Blackstone, Alyssa
Carter, Lauren
Starr, Tyler N.
Greaney, Allison J.
Lee, Kelly K.
Veesler, David
Bloom, Jesse D.
King, Neil P.
author_facet Ellis, Daniel
Brunette, Natalie
Crawford, Katharine H. D.
Walls, Alexandra C.
Pham, Minh N.
Chen, Chengbo
Herpoldt, Karla-Luise
Fiala, Brooke
Murphy, Michael
Pettie, Deleah
Kraft, John C.
Malone, Keara D.
Navarro, Mary Jane
Ogohara, Cassandra
Kepl, Elizabeth
Ravichandran, Rashmi
Sydeman, Claire
Ahlrichs, Maggie
Johnson, Max
Blackstone, Alyssa
Carter, Lauren
Starr, Tyler N.
Greaney, Allison J.
Lee, Kelly K.
Veesler, David
Bloom, Jesse D.
King, Neil P.
author_sort Ellis, Daniel
collection PubMed
description The unprecedented global demand for SARS-CoV-2 vaccines has demonstrated the need for highly effective vaccine candidates that are thermostable and amenable to large-scale manufacturing. Nanoparticle immunogens presenting the receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein (S) in repetitive arrays are being advanced as second-generation vaccine candidates, as they feature robust manufacturing characteristics and have shown promising immunogenicity in preclinical models. Here, we used previously reported deep mutational scanning (DMS) data to guide the design of stabilized variants of the RBD. The selected mutations fill a cavity in the RBD that has been identified as a linoleic acid binding pocket. Screening of several designs led to the selection of two lead candidates that expressed at higher yields than the wild-type RBD. These stabilized RBDs possess enhanced thermal stability and resistance to aggregation, particularly when incorporated into an icosahedral nanoparticle immunogen that maintained its integrity and antigenicity for 28 days at 35-40°C, while corresponding immunogens displaying the wild-type RBD experienced aggregation and loss of antigenicity. The stabilized immunogens preserved the potent immunogenicity of the original nanoparticle immunogen, which is currently being evaluated in a Phase I/II clinical trial. Our findings may improve the scalability and stability of RBD-based coronavirus vaccines in any format and more generally highlight the utility of comprehensive DMS data in guiding vaccine design.
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spelling pubmed-82766962021-07-14 Stabilization of the SARS-CoV-2 Spike Receptor-Binding Domain Using Deep Mutational Scanning and Structure-Based Design Ellis, Daniel Brunette, Natalie Crawford, Katharine H. D. Walls, Alexandra C. Pham, Minh N. Chen, Chengbo Herpoldt, Karla-Luise Fiala, Brooke Murphy, Michael Pettie, Deleah Kraft, John C. Malone, Keara D. Navarro, Mary Jane Ogohara, Cassandra Kepl, Elizabeth Ravichandran, Rashmi Sydeman, Claire Ahlrichs, Maggie Johnson, Max Blackstone, Alyssa Carter, Lauren Starr, Tyler N. Greaney, Allison J. Lee, Kelly K. Veesler, David Bloom, Jesse D. King, Neil P. Front Immunol Immunology The unprecedented global demand for SARS-CoV-2 vaccines has demonstrated the need for highly effective vaccine candidates that are thermostable and amenable to large-scale manufacturing. Nanoparticle immunogens presenting the receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein (S) in repetitive arrays are being advanced as second-generation vaccine candidates, as they feature robust manufacturing characteristics and have shown promising immunogenicity in preclinical models. Here, we used previously reported deep mutational scanning (DMS) data to guide the design of stabilized variants of the RBD. The selected mutations fill a cavity in the RBD that has been identified as a linoleic acid binding pocket. Screening of several designs led to the selection of two lead candidates that expressed at higher yields than the wild-type RBD. These stabilized RBDs possess enhanced thermal stability and resistance to aggregation, particularly when incorporated into an icosahedral nanoparticle immunogen that maintained its integrity and antigenicity for 28 days at 35-40°C, while corresponding immunogens displaying the wild-type RBD experienced aggregation and loss of antigenicity. The stabilized immunogens preserved the potent immunogenicity of the original nanoparticle immunogen, which is currently being evaluated in a Phase I/II clinical trial. Our findings may improve the scalability and stability of RBD-based coronavirus vaccines in any format and more generally highlight the utility of comprehensive DMS data in guiding vaccine design. Frontiers Media S.A. 2021-06-29 /pmc/articles/PMC8276696/ /pubmed/34267764 http://dx.doi.org/10.3389/fimmu.2021.710263 Text en Copyright © 2021 Ellis, Brunette, Crawford, Walls, Pham, Chen, Herpoldt, Fiala, Murphy, Pettie, Kraft, Malone, Navarro, Ogohara, Kepl, Ravichandran, Sydeman, Ahlrichs, Johnson, Blackstone, Carter, Starr, Greaney, Lee, Veesler, Bloom and King https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Ellis, Daniel
Brunette, Natalie
Crawford, Katharine H. D.
Walls, Alexandra C.
Pham, Minh N.
Chen, Chengbo
Herpoldt, Karla-Luise
Fiala, Brooke
Murphy, Michael
Pettie, Deleah
Kraft, John C.
Malone, Keara D.
Navarro, Mary Jane
Ogohara, Cassandra
Kepl, Elizabeth
Ravichandran, Rashmi
Sydeman, Claire
Ahlrichs, Maggie
Johnson, Max
Blackstone, Alyssa
Carter, Lauren
Starr, Tyler N.
Greaney, Allison J.
Lee, Kelly K.
Veesler, David
Bloom, Jesse D.
King, Neil P.
Stabilization of the SARS-CoV-2 Spike Receptor-Binding Domain Using Deep Mutational Scanning and Structure-Based Design
title Stabilization of the SARS-CoV-2 Spike Receptor-Binding Domain Using Deep Mutational Scanning and Structure-Based Design
title_full Stabilization of the SARS-CoV-2 Spike Receptor-Binding Domain Using Deep Mutational Scanning and Structure-Based Design
title_fullStr Stabilization of the SARS-CoV-2 Spike Receptor-Binding Domain Using Deep Mutational Scanning and Structure-Based Design
title_full_unstemmed Stabilization of the SARS-CoV-2 Spike Receptor-Binding Domain Using Deep Mutational Scanning and Structure-Based Design
title_short Stabilization of the SARS-CoV-2 Spike Receptor-Binding Domain Using Deep Mutational Scanning and Structure-Based Design
title_sort stabilization of the sars-cov-2 spike receptor-binding domain using deep mutational scanning and structure-based design
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8276696/
https://www.ncbi.nlm.nih.gov/pubmed/34267764
http://dx.doi.org/10.3389/fimmu.2021.710263
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