Serological Test to Determine Exposure to SARS-CoV-2: ELISA Based on the Receptor-Binding Domain of the Spike Protein (S-RBD<sub>N318-V510</sub>) Expressed in <i>Escherichia coli</i>
Massive worldwide serological testing for SARS-CoV-2 is needed to determine the extent of virus exposure in a particular region, the ratio of symptomatic to asymptomatic infected persons, and the duration and extent of immunity after infection. To achieve this, the development and production of reli...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-02-01
|
| Series: | Diagnostics |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2075-4418/11/2/271 |
| _version_ | 1797411998494883840 |
|---|---|
| author | Alan Roberto Márquez-Ipiña Everardo González-González Iram Pablo Rodríguez-Sánchez Itzel Montserrat Lara-Mayorga Luis Alberto Mejía-Manzano Mónica Gabriela Sánchez-Salazar José Guillermo González-Valdez Rocio Ortiz-López Augusto Rojas-Martínez Grissel Trujillo-de Santiago Mario Moisés Alvarez |
| author_facet | Alan Roberto Márquez-Ipiña Everardo González-González Iram Pablo Rodríguez-Sánchez Itzel Montserrat Lara-Mayorga Luis Alberto Mejía-Manzano Mónica Gabriela Sánchez-Salazar José Guillermo González-Valdez Rocio Ortiz-López Augusto Rojas-Martínez Grissel Trujillo-de Santiago Mario Moisés Alvarez |
| author_sort | Alan Roberto Márquez-Ipiña |
| collection | DOAJ |
| description | Massive worldwide serological testing for SARS-CoV-2 is needed to determine the extent of virus exposure in a particular region, the ratio of symptomatic to asymptomatic infected persons, and the duration and extent of immunity after infection. To achieve this, the development and production of reliable and cost-effective SARS-CoV-2 antigens is critical. We report the bacterial production of the peptide S-RBD<sub>N318-V510</sub>, which contains the receptor-binding domain of the SARS-CoV-2 spike protein (region of 193 amino acid residues from asparagine-318 to valine-510) of the SARS-CoV-2 spike protein. We purified this peptide using a straightforward approach involving bacterial lysis, his-tag-mediated affinity chromatography, and imidazole-assisted refolding. The antigen performances of S-RBD<sub>N318-V510</sub> and a commercial full-length spike protein were compared in ELISAs. In direct ELISAs, where the antigen was directly bound to the ELISA surface, both antigens discriminated sera from non-exposed and exposed individuals. However, the discriminating resolution was better in ELISAs that used the full-spike antigen than the S-RBD<sub>N318-V510</sub>. Attachment of the antigens to the ELISA surface using a layer of anti-histidine antibodies gave equivalent resolution for both S-RBD<sub>N318-V510</sub> and the full-length spike protein. Results demonstrate that ELISA-functional SARS-CoV-2 antigens can be produced in bacterial cultures, and that S-RBD<sub>N318-V510</sub> may represent a cost-effective alternative to the use of structurally more complex antigens in serological COVID-19 testing. |
| first_indexed | 2024-03-09T04:54:40Z |
| format | Article |
| id | doaj.art-f0f993e213fd4183ba2bfbd55f71991b |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-09T04:54:40Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-f0f993e213fd4183ba2bfbd55f71991b2023-12-03T13:07:14ZengMDPI AGDiagnostics2075-44182021-02-0111227110.3390/diagnostics11020271Serological Test to Determine Exposure to SARS-CoV-2: ELISA Based on the Receptor-Binding Domain of the Spike Protein (S-RBD<sub>N318-V510</sub>) Expressed in <i>Escherichia coli</i>Alan Roberto Márquez-Ipiña0Everardo González-González1Iram Pablo Rodríguez-Sánchez2Itzel Montserrat Lara-Mayorga3Luis Alberto Mejía-Manzano4Mónica Gabriela Sánchez-Salazar5José Guillermo González-Valdez6Rocio Ortiz-López7Augusto Rojas-Martínez8Grissel Trujillo-de Santiago9Mario Moisés Alvarez10Centro de Biotecnología-FEMSA, Tecnologico de Monterrey, Monterrey CP 64849, NL, MexicoDepartamento de Bioingeniería, Tecnologico de Monterrey, Monterrey CP 64849, NL, MexicoLaboratorio de Fisiología Molecular y Estructural, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, San Nicolas de los Garza CP 66455, NL, MexicoCentro de Biotecnología-FEMSA, Tecnologico de Monterrey, Monterrey CP 64849, NL, MexicoCentro de Biotecnología-FEMSA, Tecnologico de Monterrey, Monterrey CP 64849, NL, MexicoDepartamento de Bioingeniería, Tecnologico de Monterrey, Monterrey CP 64849, NL, MexicoCentro de Biotecnología-FEMSA, Tecnologico de Monterrey, Monterrey CP 64849, NL, MexicoTecnologico de Monterrey, Escuela de Medicina y Ciencias de la Salud, Monterrey CP 64718, NL, MexicoTecnologico de Monterrey, Escuela de Medicina y Ciencias de la Salud, Monterrey CP 64718, NL, MexicoCentro de Biotecnología-FEMSA, Tecnologico de Monterrey, Monterrey CP 64849, NL, MexicoCentro de Biotecnología-FEMSA, Tecnologico de Monterrey, Monterrey CP 64849, NL, MexicoMassive worldwide serological testing for SARS-CoV-2 is needed to determine the extent of virus exposure in a particular region, the ratio of symptomatic to asymptomatic infected persons, and the duration and extent of immunity after infection. To achieve this, the development and production of reliable and cost-effective SARS-CoV-2 antigens is critical. We report the bacterial production of the peptide S-RBD<sub>N318-V510</sub>, which contains the receptor-binding domain of the SARS-CoV-2 spike protein (region of 193 amino acid residues from asparagine-318 to valine-510) of the SARS-CoV-2 spike protein. We purified this peptide using a straightforward approach involving bacterial lysis, his-tag-mediated affinity chromatography, and imidazole-assisted refolding. The antigen performances of S-RBD<sub>N318-V510</sub> and a commercial full-length spike protein were compared in ELISAs. In direct ELISAs, where the antigen was directly bound to the ELISA surface, both antigens discriminated sera from non-exposed and exposed individuals. However, the discriminating resolution was better in ELISAs that used the full-spike antigen than the S-RBD<sub>N318-V510</sub>. Attachment of the antigens to the ELISA surface using a layer of anti-histidine antibodies gave equivalent resolution for both S-RBD<sub>N318-V510</sub> and the full-length spike protein. Results demonstrate that ELISA-functional SARS-CoV-2 antigens can be produced in bacterial cultures, and that S-RBD<sub>N318-V510</sub> may represent a cost-effective alternative to the use of structurally more complex antigens in serological COVID-19 testing.https://www.mdpi.com/2075-4418/11/2/271SARS-CoV-2COVID-19ELISAserological testingspikereceptor binding domain |
| spellingShingle | Alan Roberto Márquez-Ipiña Everardo González-González Iram Pablo Rodríguez-Sánchez Itzel Montserrat Lara-Mayorga Luis Alberto Mejía-Manzano Mónica Gabriela Sánchez-Salazar José Guillermo González-Valdez Rocio Ortiz-López Augusto Rojas-Martínez Grissel Trujillo-de Santiago Mario Moisés Alvarez Serological Test to Determine Exposure to SARS-CoV-2: ELISA Based on the Receptor-Binding Domain of the Spike Protein (S-RBD<sub>N318-V510</sub>) Expressed in <i>Escherichia coli</i> Diagnostics SARS-CoV-2 COVID-19 ELISA serological testing spike receptor binding domain |
| title | Serological Test to Determine Exposure to SARS-CoV-2: ELISA Based on the Receptor-Binding Domain of the Spike Protein (S-RBD<sub>N318-V510</sub>) Expressed in <i>Escherichia coli</i> |
| title_full | Serological Test to Determine Exposure to SARS-CoV-2: ELISA Based on the Receptor-Binding Domain of the Spike Protein (S-RBD<sub>N318-V510</sub>) Expressed in <i>Escherichia coli</i> |
| title_fullStr | Serological Test to Determine Exposure to SARS-CoV-2: ELISA Based on the Receptor-Binding Domain of the Spike Protein (S-RBD<sub>N318-V510</sub>) Expressed in <i>Escherichia coli</i> |
| title_full_unstemmed | Serological Test to Determine Exposure to SARS-CoV-2: ELISA Based on the Receptor-Binding Domain of the Spike Protein (S-RBD<sub>N318-V510</sub>) Expressed in <i>Escherichia coli</i> |
| title_short | Serological Test to Determine Exposure to SARS-CoV-2: ELISA Based on the Receptor-Binding Domain of the Spike Protein (S-RBD<sub>N318-V510</sub>) Expressed in <i>Escherichia coli</i> |
| title_sort | serological test to determine exposure to sars cov 2 elisa based on the receptor binding domain of the spike protein s rbd sub n318 v510 sub expressed in i escherichia coli i |
| topic | SARS-CoV-2 COVID-19 ELISA serological testing spike receptor binding domain |
| url | https://www.mdpi.com/2075-4418/11/2/271 |
| work_keys_str_mv | AT alanrobertomarquezipina serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT everardogonzalezgonzalez serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT irampablorodriguezsanchez serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT itzelmontserratlaramayorga serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT luisalbertomejiamanzano serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT monicagabrielasanchezsalazar serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT joseguillermogonzalezvaldez serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT rocioortizlopez serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT augustorojasmartinez serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT grisseltrujillodesantiago serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii AT mariomoisesalvarez serologicaltesttodetermineexposuretosarscov2elisabasedonthereceptorbindingdomainofthespikeproteinsrbdsubn318v510subexpressediniescherichiacolii |