An Engineered Receptor-Binding Domain Improves the Immunogenicity of Multivalent SARS-CoV-2 Vaccines
Copyright © 2021 Guo et al..
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein mediates viral entry into cells expressing angiotensin-converting enzyme 2 (ACE2). The S protein engages ACE2 through its receptor-binding domain (RBD), an independently folded 197-amino-acid fragment of the 1,273-amino-acid S-protein protomer. The RBD is the primary SARS-CoV-2 neutralizing epitope and a critical target of any SARS-CoV-2 vaccine. Here, we show that this RBD conjugated to each of two carrier proteins elicited more potent neutralizing responses in immunized rodents than did a similarly conjugated proline-stabilized S-protein ectodomain. Nonetheless, the native RBD is expressed inefficiently, limiting its usefulness as a vaccine antigen. However, we show that an RBD engineered with four novel glycosylation sites (gRBD) is expressed markedly more efficiently and generates a more potent neutralizing responses as a DNA vaccine antigen than the wild-type RBD or the full-length S protein, especially when fused to multivalent carriers, such as a Helicobacter pylori ferritin 24-mer. Further, gRBD is more immunogenic than the wild-type RBD when administered as a subunit protein vaccine. Our data suggest that multivalent gRBD antigens can reduce costs and doses, and improve the immunogenicity, of all major classes of SARS-CoV-2 vaccines.IMPORTANCE All available vaccines for coronavirus disease 2019 (COVID-19) express or deliver the full-length SARS-CoV-2 spike (S) protein. We show that this antigen is not optimal, consistent with observations that the vast majority of the neutralizing response to the virus is focused on the S-protein receptor-binding domain (RBD). However, this RBD is not expressed well as an independent domain, especially when expressed as a fusion protein with a multivalent scaffold. We therefore engineered a more highly expressed form of the SARS-CoV-2 RBD by introducing four glycosylation sites into a face of the RBD normally occluded in the full S protein. We show that this engineered protein, gRBD, is more immunogenic than the wild-type RBD or the full-length S protein in both genetic and protein-delivered vaccines.
| Errataetall: | |
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| Medienart: |
E-Artikel |
| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:12 |
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| Enthalten in: |
mBio - 12(2021), 3 vom: 11. Mai |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Guo, Yan [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 24.05.2021 Date Revised 25.07.2021 published: Electronic UpdateOf: bioRxiv. 2020 Nov 18;:. - PMID 33236008 Citation Status MEDLINE |
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| doi: |
10.1128/mBio.00930-21 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
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| 100 | 1 | |a Guo, Yan |e verfasserin |4 aut | |
| 245 | 1 | 3 | |a An Engineered Receptor-Binding Domain Improves the Immunogenicity of Multivalent SARS-CoV-2 Vaccines |
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| 500 | |a Date Revised 25.07.2021 | ||
| 500 | |a published: Electronic | ||
| 500 | |a UpdateOf: bioRxiv. 2020 Nov 18;:. - PMID 33236008 | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2021 Guo et al. | ||
| 520 | |a The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein mediates viral entry into cells expressing angiotensin-converting enzyme 2 (ACE2). The S protein engages ACE2 through its receptor-binding domain (RBD), an independently folded 197-amino-acid fragment of the 1,273-amino-acid S-protein protomer. The RBD is the primary SARS-CoV-2 neutralizing epitope and a critical target of any SARS-CoV-2 vaccine. Here, we show that this RBD conjugated to each of two carrier proteins elicited more potent neutralizing responses in immunized rodents than did a similarly conjugated proline-stabilized S-protein ectodomain. Nonetheless, the native RBD is expressed inefficiently, limiting its usefulness as a vaccine antigen. However, we show that an RBD engineered with four novel glycosylation sites (gRBD) is expressed markedly more efficiently and generates a more potent neutralizing responses as a DNA vaccine antigen than the wild-type RBD or the full-length S protein, especially when fused to multivalent carriers, such as a Helicobacter pylori ferritin 24-mer. Further, gRBD is more immunogenic than the wild-type RBD when administered as a subunit protein vaccine. Our data suggest that multivalent gRBD antigens can reduce costs and doses, and improve the immunogenicity, of all major classes of SARS-CoV-2 vaccines.IMPORTANCE All available vaccines for coronavirus disease 2019 (COVID-19) express or deliver the full-length SARS-CoV-2 spike (S) protein. We show that this antigen is not optimal, consistent with observations that the vast majority of the neutralizing response to the virus is focused on the S-protein receptor-binding domain (RBD). However, this RBD is not expressed well as an independent domain, especially when expressed as a fusion protein with a multivalent scaffold. We therefore engineered a more highly expressed form of the SARS-CoV-2 RBD by introducing four glycosylation sites into a face of the RBD normally occluded in the full S protein. We show that this engineered protein, gRBD, is more immunogenic than the wild-type RBD or the full-length S protein in both genetic and protein-delivered vaccines | ||
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| 650 | 7 | |a Receptors, Coronavirus |2 NLM | |
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| 650 | 7 | |a Vaccines, Conjugate |2 NLM | |
| 650 | 7 | |a Vaccines, Synthetic |2 NLM | |
| 650 | 7 | |a spike protein, SARS-CoV-2 |2 NLM | |
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| 700 | 1 | |a He, Wenhui |e verfasserin |4 aut | |
| 700 | 1 | |a Mou, Huihui |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Lizhou |e verfasserin |4 aut | |
| 700 | 1 | |a Chang, Jing |e verfasserin |4 aut | |
| 700 | 1 | |a Peng, Shoujiao |e verfasserin |4 aut | |
| 700 | 1 | |a Ojha, Amrita |e verfasserin |4 aut | |
| 700 | 1 | |a Tavora, Rubens |e verfasserin |4 aut | |
| 700 | 1 | |a Parcells, Mark S |e verfasserin |4 aut | |
| 700 | 1 | |a Luo, Guangxiang |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Wenhui |e verfasserin |4 aut | |
| 700 | 1 | |a Zhong, Guocai |e verfasserin |4 aut | |
| 700 | 1 | |a Choe, Hyeryun |e verfasserin |4 aut | |
| 700 | 1 | |a Farzan, Michael |e verfasserin |4 aut | |
| 700 | 1 | |a Quinlan, Brian D |e verfasserin |4 aut | |
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