The C-Terminal Half of SARS-CoV-2 Nucleocapsid Protein, Industrially Produced in Plants, Is Valid as Antigen in COVID-19 Serological Tests
BackgroundThe fight against the current coronavirus disease 2019 (COVID-19) pandemic has created a huge demand of biotechnological, pharmaceutical, research and sanitary materials at unprecedented scales. One of the most urgent demands affects the diagnostic tests. The growing need for rapid and acc...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2021-07-01
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| Series: | Frontiers in Plant Science |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2021.699665/full |
| _version_ | 1818651490579906560 |
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| author | Laura Williams Silvia Jurado Francisco Llorente Alberto Romualdo Sara González Ana Saconne Isabel Bronchalo Mercedes Martínez-Cortes Beatriz Pérez-Gómez Beatriz Pérez-Gómez Fernando Ponz Miguel Ángel Jiménez-Clavero Miguel Ángel Jiménez-Clavero Pablo Lunello |
| author_facet | Laura Williams Silvia Jurado Francisco Llorente Alberto Romualdo Sara González Ana Saconne Isabel Bronchalo Mercedes Martínez-Cortes Beatriz Pérez-Gómez Beatriz Pérez-Gómez Fernando Ponz Miguel Ángel Jiménez-Clavero Miguel Ángel Jiménez-Clavero Pablo Lunello |
| author_sort | Laura Williams |
| collection | DOAJ |
| description | BackgroundThe fight against the current coronavirus disease 2019 (COVID-19) pandemic has created a huge demand of biotechnological, pharmaceutical, research and sanitary materials at unprecedented scales. One of the most urgent demands affects the diagnostic tests. The growing need for rapid and accurate laboratory diagnostic tests requires the development of biotechnological processes aimed at producing reagents able to cope with this demand in a scalable, cost-effective manner, with rapid turnaround times. This is particularly applicable to the antigens employed in serological tests. Recombinant protein expression using plants as biofactories is particularly suitable for mass production of protein antigens useful in serological diagnosis, with a neat advantage in economic terms.MethodsWe expressed a large portion of the nucleoprotein (N) derived from SARS-CoV-2 in Nicotiana benthamiana plants. After purification, the recombinant N protein obtained was used to develop an indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to SARS-CoV-2 in human sera. To validate the ELISA, a panel of 416 sera from exposed personnel at essential services in Madrid City Council were tested, and the results compared to those obtained by another ELISA, already validated, used as reference. Furthermore, a subset of samples for which RT-PCR results were available were used to confirm sensitivity and specificity of the test.ResultsThe performance of the N protein expressed in plants as antigen in serologic test for SARS-CoV-2 antibody detection was shown to be highly satisfactory, with calculated diagnostic sensitivity of 96.41% (95% CI: 93.05–98.44) and diagnostic specificity of 96.37 (95% CI: 93.05–98.44) as compared to the reference ELISA, with a kappa (K) value of 0.928 (95% CI:0.892–0.964). Furthermore, the ELISA developed with plant-derived N antigen detected SARS-CoV-2 antibodies in 84 out of 93 sera from individuals showing RT-PCR positive results (86/93 for the reference ELISA).ConclusionThis study demonstrates that the N protein part derived from SARS-CoV-2 expressed in plants performs as a perfectly valid antigen for use in COVID-19 diagnosis. Furthermore, our results support the use of this plant platform for expression of recombinant proteins as reagents for COVID-19 diagnosis. This platform stands out as a convenient and advantageous production system, fit-for-purpose to cope with the current demand of this type of biologicals in a cost-effective manner, making diagnostic kits more affordable. |
| first_indexed | 2024-12-17T02:06:57Z |
| format | Article |
| id | doaj.art-c0acbdd268a64b82ba169d52c3676c25 |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-12-17T02:06:57Z |
| publishDate | 2021-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-c0acbdd268a64b82ba169d52c3676c252022-12-21T22:07:40ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2021-07-011210.3389/fpls.2021.699665699665The C-Terminal Half of SARS-CoV-2 Nucleocapsid Protein, Industrially Produced in Plants, Is Valid as Antigen in COVID-19 Serological TestsLaura Williams0Silvia Jurado1Francisco Llorente2Alberto Romualdo3Sara González4Ana Saconne5Isabel Bronchalo6Mercedes Martínez-Cortes7Beatriz Pérez-Gómez8Beatriz Pérez-Gómez9Fernando Ponz10Miguel Ángel Jiménez-Clavero11Miguel Ángel Jiménez-Clavero12Pablo Lunello13Agrenvec S.L., Madrid, SpainAgrenvec S.L., Madrid, SpainCentro de Investigación en Sanidad Animal (CISA), Centro Nacional Instituto de Investigación y Tecnología Agraria y Alimentaria (INIA-CSIC), Valdeolmos, SpainAgrenvec S.L., Madrid, SpainAgrenvec S.L., Madrid, SpainAgrenvec S.L., Madrid, SpainAgrenvec S.L., Madrid, SpainMadrid Salud: Public Health Services of Madrid City Council, Madrid, SpainNational Centre for Epidemiology, Instituto de Salud Carlos III, Madrid, SpainConsortium for Biomedical Research in Epidemiology and Public Health (CIBERESP), Madrid, SpainCentro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid-Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (CBGP, UPM-INIA, CSIC), Madrid, SpainCentro de Investigación en Sanidad Animal (CISA), Centro Nacional Instituto de Investigación y Tecnología Agraria y Alimentaria (INIA-CSIC), Valdeolmos, SpainConsortium for Biomedical Research in Epidemiology and Public Health (CIBERESP), Madrid, SpainAgrenvec S.L., Madrid, SpainBackgroundThe fight against the current coronavirus disease 2019 (COVID-19) pandemic has created a huge demand of biotechnological, pharmaceutical, research and sanitary materials at unprecedented scales. One of the most urgent demands affects the diagnostic tests. The growing need for rapid and accurate laboratory diagnostic tests requires the development of biotechnological processes aimed at producing reagents able to cope with this demand in a scalable, cost-effective manner, with rapid turnaround times. This is particularly applicable to the antigens employed in serological tests. Recombinant protein expression using plants as biofactories is particularly suitable for mass production of protein antigens useful in serological diagnosis, with a neat advantage in economic terms.MethodsWe expressed a large portion of the nucleoprotein (N) derived from SARS-CoV-2 in Nicotiana benthamiana plants. After purification, the recombinant N protein obtained was used to develop an indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to SARS-CoV-2 in human sera. To validate the ELISA, a panel of 416 sera from exposed personnel at essential services in Madrid City Council were tested, and the results compared to those obtained by another ELISA, already validated, used as reference. Furthermore, a subset of samples for which RT-PCR results were available were used to confirm sensitivity and specificity of the test.ResultsThe performance of the N protein expressed in plants as antigen in serologic test for SARS-CoV-2 antibody detection was shown to be highly satisfactory, with calculated diagnostic sensitivity of 96.41% (95% CI: 93.05–98.44) and diagnostic specificity of 96.37 (95% CI: 93.05–98.44) as compared to the reference ELISA, with a kappa (K) value of 0.928 (95% CI:0.892–0.964). Furthermore, the ELISA developed with plant-derived N antigen detected SARS-CoV-2 antibodies in 84 out of 93 sera from individuals showing RT-PCR positive results (86/93 for the reference ELISA).ConclusionThis study demonstrates that the N protein part derived from SARS-CoV-2 expressed in plants performs as a perfectly valid antigen for use in COVID-19 diagnosis. Furthermore, our results support the use of this plant platform for expression of recombinant proteins as reagents for COVID-19 diagnosis. This platform stands out as a convenient and advantageous production system, fit-for-purpose to cope with the current demand of this type of biologicals in a cost-effective manner, making diagnostic kits more affordable.https://www.frontiersin.org/articles/10.3389/fpls.2021.699665/fullCOVID-19SARS CoV-2 nucleocapsid proteinbiofactoriesplant expressionmolecular farmingELISA coronavirus |
| spellingShingle | Laura Williams Silvia Jurado Francisco Llorente Alberto Romualdo Sara González Ana Saconne Isabel Bronchalo Mercedes Martínez-Cortes Beatriz Pérez-Gómez Beatriz Pérez-Gómez Fernando Ponz Miguel Ángel Jiménez-Clavero Miguel Ángel Jiménez-Clavero Pablo Lunello The C-Terminal Half of SARS-CoV-2 Nucleocapsid Protein, Industrially Produced in Plants, Is Valid as Antigen in COVID-19 Serological Tests Frontiers in Plant Science COVID-19 SARS CoV-2 nucleocapsid protein biofactories plant expression molecular farming ELISA coronavirus |
| title | The C-Terminal Half of SARS-CoV-2 Nucleocapsid Protein, Industrially Produced in Plants, Is Valid as Antigen in COVID-19 Serological Tests |
| title_full | The C-Terminal Half of SARS-CoV-2 Nucleocapsid Protein, Industrially Produced in Plants, Is Valid as Antigen in COVID-19 Serological Tests |
| title_fullStr | The C-Terminal Half of SARS-CoV-2 Nucleocapsid Protein, Industrially Produced in Plants, Is Valid as Antigen in COVID-19 Serological Tests |
| title_full_unstemmed | The C-Terminal Half of SARS-CoV-2 Nucleocapsid Protein, Industrially Produced in Plants, Is Valid as Antigen in COVID-19 Serological Tests |
| title_short | The C-Terminal Half of SARS-CoV-2 Nucleocapsid Protein, Industrially Produced in Plants, Is Valid as Antigen in COVID-19 Serological Tests |
| title_sort | c terminal half of sars cov 2 nucleocapsid protein industrially produced in plants is valid as antigen in covid 19 serological tests |
| topic | COVID-19 SARS CoV-2 nucleocapsid protein biofactories plant expression molecular farming ELISA coronavirus |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2021.699665/full |
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