Recombinant Antigens Based on Non-Glycosylated Regions from RBD SARS-CoV-2 as Potential Vaccine Candidates against COVID-19
The Receptor-Binding Domain (RBD) of the Spike (S) protein from Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has glycosylation sites which can limit the production of reliable antigens expressed in prokaryotic platforms, due to glycan-mediated evasion of the host immune response. How...
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MDPI AG
2021-08-01
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| Series: | Vaccines |
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| Online Access: | https://www.mdpi.com/2076-393X/9/8/928 |
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| author | Leandro Núñez-Muñoz Gabriel Marcelino-Pérez Berenice Calderón-Pérez Miriam Pérez-Saldívar Karla Acosta-Virgen Hugo González-Conchillos Brenda Vargas-Hernández Ana Olivares-Martínez Roberto Ruiz-Medrano Daniela Roa-Velázquez Edgar Morales-Ríos Jorge Ramos-Flores Gustavo Torres-Franco Diana Peláez-González Jorge Fernández-Hernández Martha Espinosa-Cantellano Diana Tapia-Sidas José Abrahan Ramírez-Pool América Padilla-Viveros Beatriz Xoconostle-Cázares |
| author_facet | Leandro Núñez-Muñoz Gabriel Marcelino-Pérez Berenice Calderón-Pérez Miriam Pérez-Saldívar Karla Acosta-Virgen Hugo González-Conchillos Brenda Vargas-Hernández Ana Olivares-Martínez Roberto Ruiz-Medrano Daniela Roa-Velázquez Edgar Morales-Ríos Jorge Ramos-Flores Gustavo Torres-Franco Diana Peláez-González Jorge Fernández-Hernández Martha Espinosa-Cantellano Diana Tapia-Sidas José Abrahan Ramírez-Pool América Padilla-Viveros Beatriz Xoconostle-Cázares |
| author_sort | Leandro Núñez-Muñoz |
| collection | DOAJ |
| description | The Receptor-Binding Domain (RBD) of the Spike (S) protein from Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has glycosylation sites which can limit the production of reliable antigens expressed in prokaryotic platforms, due to glycan-mediated evasion of the host immune response. However, protein regions without glycosylated residues capable of inducing neutralizing antibodies could be useful for antigen production in systems that do not carry the glycosylation machinery. To test this hypothesis, the potential antigens NG06 and NG19, located within the non-glycosylated S-RBD region, were selected and expressed in <i>Escherichia coli</i>, purified by FPLC and employed to determine their immunogenic potential through detection of antibodies in serum from immunized rabbits, mice, and COVID-19 patients. IgG antibodies from sera of COVID-19-recovered patients detected the recombinant antigens NG06 and NG19 (A<sub>450 nm</sub> = 0.80 ± 0.33; 1.13 ± 0.33; and 0.11 ± 0.08 for and negatives controls, respectively). Also, the purified antigens were able to raise polyclonal antibodies in animal models evoking a strong immune response with neutralizing activity in mice model. This research highlights the usefulness of antigens based on the non-N-glycosylated region of RBD from SARS-CoV-2 for candidate vaccine development. |
| first_indexed | 2024-03-10T08:18:46Z |
| format | Article |
| id | doaj.art-9334bd9b0c7642c7a44b08eefd119a42 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-10T08:18:46Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-9334bd9b0c7642c7a44b08eefd119a422023-11-22T10:08:10ZengMDPI AGVaccines2076-393X2021-08-019892810.3390/vaccines9080928Recombinant Antigens Based on Non-Glycosylated Regions from RBD SARS-CoV-2 as Potential Vaccine Candidates against COVID-19Leandro Núñez-Muñoz0Gabriel Marcelino-Pérez1Berenice Calderón-Pérez2Miriam Pérez-Saldívar3Karla Acosta-Virgen4Hugo González-Conchillos5Brenda Vargas-Hernández6Ana Olivares-Martínez7Roberto Ruiz-Medrano8Daniela Roa-Velázquez9Edgar Morales-Ríos10Jorge Ramos-Flores11Gustavo Torres-Franco12Diana Peláez-González13Jorge Fernández-Hernández14Martha Espinosa-Cantellano15Diana Tapia-Sidas16José Abrahan Ramírez-Pool17América Padilla-Viveros18Beatriz Xoconostle-Cázares19Department of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Infectomics and Molecular Pathogenesis, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Infectomics and Molecular Pathogenesis, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Infectomics and Molecular Pathogenesis, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDoctoral Program in Nanosciences and Nanotechnology, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Biochemistry, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoLaboratory Animal Production and Experimentation Unit, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoLaboratory Animal Production and Experimentation Unit, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoLaboratory Animal Production and Experimentation Unit, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoLaboratory Animal Production and Experimentation Unit, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Infectomics and Molecular Pathogenesis, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoTransdisciplinary Doctoral Program in Scientific and Technological Development for Society, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoDepartment of Biotechnology and Bioengineering, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Av. Instituto Politécnico Nacional 2508, México City 07360, MexicoThe Receptor-Binding Domain (RBD) of the Spike (S) protein from Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has glycosylation sites which can limit the production of reliable antigens expressed in prokaryotic platforms, due to glycan-mediated evasion of the host immune response. However, protein regions without glycosylated residues capable of inducing neutralizing antibodies could be useful for antigen production in systems that do not carry the glycosylation machinery. To test this hypothesis, the potential antigens NG06 and NG19, located within the non-glycosylated S-RBD region, were selected and expressed in <i>Escherichia coli</i>, purified by FPLC and employed to determine their immunogenic potential through detection of antibodies in serum from immunized rabbits, mice, and COVID-19 patients. IgG antibodies from sera of COVID-19-recovered patients detected the recombinant antigens NG06 and NG19 (A<sub>450 nm</sub> = 0.80 ± 0.33; 1.13 ± 0.33; and 0.11 ± 0.08 for and negatives controls, respectively). Also, the purified antigens were able to raise polyclonal antibodies in animal models evoking a strong immune response with neutralizing activity in mice model. This research highlights the usefulness of antigens based on the non-N-glycosylated region of RBD from SARS-CoV-2 for candidate vaccine development.https://www.mdpi.com/2076-393X/9/8/928SARS-CoV-2receptor binding domainSpike proteinvaccineviral glycosylationprokaryotic expression |
| spellingShingle | Leandro Núñez-Muñoz Gabriel Marcelino-Pérez Berenice Calderón-Pérez Miriam Pérez-Saldívar Karla Acosta-Virgen Hugo González-Conchillos Brenda Vargas-Hernández Ana Olivares-Martínez Roberto Ruiz-Medrano Daniela Roa-Velázquez Edgar Morales-Ríos Jorge Ramos-Flores Gustavo Torres-Franco Diana Peláez-González Jorge Fernández-Hernández Martha Espinosa-Cantellano Diana Tapia-Sidas José Abrahan Ramírez-Pool América Padilla-Viveros Beatriz Xoconostle-Cázares Recombinant Antigens Based on Non-Glycosylated Regions from RBD SARS-CoV-2 as Potential Vaccine Candidates against COVID-19 Vaccines SARS-CoV-2 receptor binding domain Spike protein vaccine viral glycosylation prokaryotic expression |
| title | Recombinant Antigens Based on Non-Glycosylated Regions from RBD SARS-CoV-2 as Potential Vaccine Candidates against COVID-19 |
| title_full | Recombinant Antigens Based on Non-Glycosylated Regions from RBD SARS-CoV-2 as Potential Vaccine Candidates against COVID-19 |
| title_fullStr | Recombinant Antigens Based on Non-Glycosylated Regions from RBD SARS-CoV-2 as Potential Vaccine Candidates against COVID-19 |
| title_full_unstemmed | Recombinant Antigens Based on Non-Glycosylated Regions from RBD SARS-CoV-2 as Potential Vaccine Candidates against COVID-19 |
| title_short | Recombinant Antigens Based on Non-Glycosylated Regions from RBD SARS-CoV-2 as Potential Vaccine Candidates against COVID-19 |
| title_sort | recombinant antigens based on non glycosylated regions from rbd sars cov 2 as potential vaccine candidates against covid 19 |
| topic | SARS-CoV-2 receptor binding domain Spike protein vaccine viral glycosylation prokaryotic expression |
| url | https://www.mdpi.com/2076-393X/9/8/928 |
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