tetano
Editor, Senior Moderator
Vaccines (Basel)
. 2021 Sep 3;9(9):986.
doi: 10.3390/vaccines9090986.
Multiepitope Proteins for the Differential Detection of IgG Antibodies against RBD of the Spike Protein and Non-RBD Regions of SARS-CoV-2
Larissa R Gomes[SUP] 1 [/SUP], Andressa M Durans[SUP] 1 2 [/SUP], Paloma Napoleão-Pêgo[SUP] 1 [/SUP], Jessica A Waterman[SUP] 1 [/SUP], Mariana S Freitas[SUP] 1 [/SUP], Nathalia B R De Sá[SUP] 3 [/SUP], Lilian V Pereira[SUP] 4 [/SUP], Jéssica S Furtado[SUP] 4 [/SUP], Romário G Aquino[SUP] 4 [/SUP], Mario C R Machado[SUP] 5 [/SUP], Natalia Fintelman-Rodrigues[SUP] 1 6 [/SUP], Thiago M L Souza[SUP] 1 6 [/SUP], Carlos M Morel[SUP] 1 [/SUP], David W Provance[SUP] 1 2 [/SUP], Salvatore G De-Simone[SUP] 1 7 [/SUP]
Affiliations
Abstract
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.
Keywords: COVID-19; IgG; SARS-CoV-2; coronavirus; de novo design; linear B cell epitopes; serodiagnostic.
. 2021 Sep 3;9(9):986.
doi: 10.3390/vaccines9090986.
Multiepitope Proteins for the Differential Detection of IgG Antibodies against RBD of the Spike Protein and Non-RBD Regions of SARS-CoV-2
Larissa R Gomes[SUP] 1 [/SUP], Andressa M Durans[SUP] 1 2 [/SUP], Paloma Napoleão-Pêgo[SUP] 1 [/SUP], Jessica A Waterman[SUP] 1 [/SUP], Mariana S Freitas[SUP] 1 [/SUP], Nathalia B R De Sá[SUP] 3 [/SUP], Lilian V Pereira[SUP] 4 [/SUP], Jéssica S Furtado[SUP] 4 [/SUP], Romário G Aquino[SUP] 4 [/SUP], Mario C R Machado[SUP] 5 [/SUP], Natalia Fintelman-Rodrigues[SUP] 1 6 [/SUP], Thiago M L Souza[SUP] 1 6 [/SUP], Carlos M Morel[SUP] 1 [/SUP], David W Provance[SUP] 1 2 [/SUP], Salvatore G De-Simone[SUP] 1 7 [/SUP]
Affiliations
- PMID: 34579223
- DOI: 10.3390/vaccines9090986
Abstract
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.
Keywords: COVID-19; IgG; SARS-CoV-2; coronavirus; de novo design; linear B cell epitopes; serodiagnostic.