Multiepitope Proteins for the Differential Detection of IgG Antibodies against RBD of the Spike Protein and Non-RBD Regions of SARS-CoV-2
The COVID-19 pandemic has exposed the extent of global connectivity and collective vulnerability to emerging diseases. From its suspected origins in Wuhan, China, it spread to all corners of the world in a matter of months. The absence of high-performance, rapid diagnostic methods that could identify asymptomatic carriers contributed to its worldwide transmission. Serological tests offer numerous benefits compared to other assay platforms to screen large populations. First-generation assays contain targets that represent proteins from SARS-CoV-2. While they could be quickly produced, each actually has a mixture of specific and non-specific epitopes that vary in their reactivity for antibodies. To generate the next generation of the assay, epitopes were identified in three SARS-Cov-2 proteins (S, N, and Orf3a) by SPOT synthesis analysis. After their similarity to other pathogen sequences was analyzed, 11 epitopes outside of the receptor-binding domain (RBD) of the spike protein that showed high reactivity and uniqueness to the virus. These were incorporated into a ß-barrel protein core to create a highly chimeric protein. Another de novo protein was designed that contained only epitopes in the RBD. In-house ELISAs suggest that both multiepitope proteins can serve as targets for high-performance diagnostic tests. Our approach to bioengineer chimeric proteins is highly amenable to other pathogens and immunological uses.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:9 |
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| Enthalten in: |
Vaccines - 9(2021), 9 vom: 03. Sept. |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Gomes, Larissa R [VerfasserIn] |
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| Links: |
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| Themen: |
COVID-19 |
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| Anmerkungen: |
Date Revised 01.10.2021 published: Electronic Citation Status PubMed-not-MEDLINE |
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| doi: |
10.3390/vaccines9090986 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 520 | |a The COVID-19 pandemic has exposed the extent of global connectivity and collective vulnerability to emerging diseases. From its suspected origins in Wuhan, China, it spread to all corners of the world in a matter of months. The absence of high-performance, rapid diagnostic methods that could identify asymptomatic carriers contributed to its worldwide transmission. Serological tests offer numerous benefits compared to other assay platforms to screen large populations. First-generation assays contain targets that represent proteins from SARS-CoV-2. While they could be quickly produced, each actually has a mixture of specific and non-specific epitopes that vary in their reactivity for antibodies. To generate the next generation of the assay, epitopes were identified in three SARS-Cov-2 proteins (S, N, and Orf3a) by SPOT synthesis analysis. After their similarity to other pathogen sequences was analyzed, 11 epitopes outside of the receptor-binding domain (RBD) of the spike protein that showed high reactivity and uniqueness to the virus. These were incorporated into a ß-barrel protein core to create a highly chimeric protein. Another de novo protein was designed that contained only epitopes in the RBD. In-house ELISAs suggest that both multiepitope proteins can serve as targets for high-performance diagnostic tests. Our approach to bioengineer chimeric proteins is highly amenable to other pathogens and immunological uses | ||
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| 700 | 1 | |a Durans, Andressa M |e verfasserin |4 aut | |
| 700 | 1 | |a Napoleão-Pêgo, Paloma |e verfasserin |4 aut | |
| 700 | 1 | |a Waterman, Jessica A |e verfasserin |4 aut | |
| 700 | 1 | |a Freitas, Mariana S |e verfasserin |4 aut | |
| 700 | 1 | |a De Sá, Nathalia B R |e verfasserin |4 aut | |
| 700 | 1 | |a Pereira, Lilian V |e verfasserin |4 aut | |
| 700 | 1 | |a Furtado, Jéssica S |e verfasserin |4 aut | |
| 700 | 1 | |a Aquino, Romário G |e verfasserin |4 aut | |
| 700 | 1 | |a Machado, Mario C R |e verfasserin |4 aut | |
| 700 | 1 | |a Fintelman-Rodrigues, Natalia |e verfasserin |4 aut | |
| 700 | 1 | |a Souza, Thiago M L |e verfasserin |4 aut | |
| 700 | 1 | |a Morel, Carlos M |e verfasserin |4 aut | |
| 700 | 1 | |a Provance, David W |e verfasserin |4 aut | |
| 700 | 1 | |a De-Simone, Salvatore G |e verfasserin |4 aut | |
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