Inactivated and Immunogenic SARS-CoV-2 for Safe Use in Immunoassays and as an Immunization Control for Non-Clinical Trials
Successful SARS-CoV-2 inactivation allows its safe use in Biosafety Level 2 facilities, and the use of the whole viral particle helps in the development of analytical methods and a more reliable immune response, contributing to the development and improvement of in vitro and in vivo assays. In order...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-06-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/15/7/1486 |
| _version_ | 1797587227912437760 |
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| author | Mariana Pierre de Barros Gomes José Henrique Rezende Linhares Tiago Pereira dos Santos Renata Carvalho Pereira Renata Tourinho Santos Stephanie Almeida da Silva Marta Cristina de Oliveira Souza Juliana Fernandes Amorim da Silva Gisela Freitas Trindade Viviane Silva Gomes Débora Ferreira Barreto-Vieira Milena Mouta Verdan França Carvalho Ana Paula Dinis Ano Bom Noemi Rovaris Gardinali Rodrigo Müller Nathalia dos Santos Alves Luma da Cruz Moura Patrícia Cristina da Costa Neves Gabriela Santos Esteves Waleska Dias Schwarcz Sotiris Missailidis Ygara da Silva Mendes Sheila Maria Barbosa de Lima |
| author_facet | Mariana Pierre de Barros Gomes José Henrique Rezende Linhares Tiago Pereira dos Santos Renata Carvalho Pereira Renata Tourinho Santos Stephanie Almeida da Silva Marta Cristina de Oliveira Souza Juliana Fernandes Amorim da Silva Gisela Freitas Trindade Viviane Silva Gomes Débora Ferreira Barreto-Vieira Milena Mouta Verdan França Carvalho Ana Paula Dinis Ano Bom Noemi Rovaris Gardinali Rodrigo Müller Nathalia dos Santos Alves Luma da Cruz Moura Patrícia Cristina da Costa Neves Gabriela Santos Esteves Waleska Dias Schwarcz Sotiris Missailidis Ygara da Silva Mendes Sheila Maria Barbosa de Lima |
| author_sort | Mariana Pierre de Barros Gomes |
| collection | DOAJ |
| description | Successful SARS-CoV-2 inactivation allows its safe use in Biosafety Level 2 facilities, and the use of the whole viral particle helps in the development of analytical methods and a more reliable immune response, contributing to the development and improvement of in vitro and in vivo assays. In order to obtain a functional product, we evaluated several inactivation protocols and observed that 0.03% beta-propiolactone for 24 h was the best condition tested, as it promoted SARS-CoV-2 inactivation above 99.99% and no cytopathic effect was visualized after five serial passages. Moreover, RT-qPCR and transmission electron microscopy revealed that RNA quantification and viral structure integrity were preserved. The antigenicity of inactivated SARS-CoV-2 was confirmed by ELISA using different Spike-neutralizing monoclonal antibodies. K18-hACE2 mice immunized with inactivated SARS-CoV-2, formulated in AddaS0<sub>3</sub><sup>TM</sup>, presented high neutralizing antibody titers, no significant weight loss, and longer survival than controls from a lethal challenge, despite RNA detection in the oropharyngeal swab, lung, and brain. This work emphasizes the importance of using different techniques to confirm viral inactivation and avoid potentially disastrous contamination. We believe that an efficiently inactivated product can be used in several applications, including the development and improvement of molecular diagnostic kits, as an antigen for antibody production as well as a control for non-clinical trials. |
| first_indexed | 2024-03-11T00:34:08Z |
| format | Article |
| id | doaj.art-588292882b6f489cb29d6b8b26dc6815 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-11T00:34:08Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-588292882b6f489cb29d6b8b26dc68152023-11-18T21:44:22ZengMDPI AGViruses1999-49152023-06-01157148610.3390/v15071486Inactivated and Immunogenic SARS-CoV-2 for Safe Use in Immunoassays and as an Immunization Control for Non-Clinical TrialsMariana Pierre de Barros Gomes0José Henrique Rezende Linhares1Tiago Pereira dos Santos2Renata Carvalho Pereira3Renata Tourinho Santos4Stephanie Almeida da Silva5Marta Cristina de Oliveira Souza6Juliana Fernandes Amorim da Silva7Gisela Freitas Trindade8Viviane Silva Gomes9Débora Ferreira Barreto-Vieira10Milena Mouta Verdan França Carvalho11Ana Paula Dinis Ano Bom12Noemi Rovaris Gardinali13Rodrigo Müller14Nathalia dos Santos Alves15Luma da Cruz Moura16Patrícia Cristina da Costa Neves17Gabriela Santos Esteves18Waleska Dias Schwarcz19Sotiris Missailidis20Ygara da Silva Mendes21Sheila Maria Barbosa de Lima22Virological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilViral Morphology and Morphogenesis Laboratory, Oswaldo Cruz Institute/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilImmunological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilImmunological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilPre-Clinical Trials Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilImmunological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilRecombinant Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilInstitute of Technology in Immunobiologicals, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilVirological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, BrazilSuccessful SARS-CoV-2 inactivation allows its safe use in Biosafety Level 2 facilities, and the use of the whole viral particle helps in the development of analytical methods and a more reliable immune response, contributing to the development and improvement of in vitro and in vivo assays. In order to obtain a functional product, we evaluated several inactivation protocols and observed that 0.03% beta-propiolactone for 24 h was the best condition tested, as it promoted SARS-CoV-2 inactivation above 99.99% and no cytopathic effect was visualized after five serial passages. Moreover, RT-qPCR and transmission electron microscopy revealed that RNA quantification and viral structure integrity were preserved. The antigenicity of inactivated SARS-CoV-2 was confirmed by ELISA using different Spike-neutralizing monoclonal antibodies. K18-hACE2 mice immunized with inactivated SARS-CoV-2, formulated in AddaS0<sub>3</sub><sup>TM</sup>, presented high neutralizing antibody titers, no significant weight loss, and longer survival than controls from a lethal challenge, despite RNA detection in the oropharyngeal swab, lung, and brain. This work emphasizes the importance of using different techniques to confirm viral inactivation and avoid potentially disastrous contamination. We believe that an efficiently inactivated product can be used in several applications, including the development and improvement of molecular diagnostic kits, as an antigen for antibody production as well as a control for non-clinical trials.https://www.mdpi.com/1999-4915/15/7/1486SARS-CoV-2 inactivationbeta-propiolactoneserial passagesTCID<sub>50</sub>RT-qPCRnon-clinical trials |
| spellingShingle | Mariana Pierre de Barros Gomes José Henrique Rezende Linhares Tiago Pereira dos Santos Renata Carvalho Pereira Renata Tourinho Santos Stephanie Almeida da Silva Marta Cristina de Oliveira Souza Juliana Fernandes Amorim da Silva Gisela Freitas Trindade Viviane Silva Gomes Débora Ferreira Barreto-Vieira Milena Mouta Verdan França Carvalho Ana Paula Dinis Ano Bom Noemi Rovaris Gardinali Rodrigo Müller Nathalia dos Santos Alves Luma da Cruz Moura Patrícia Cristina da Costa Neves Gabriela Santos Esteves Waleska Dias Schwarcz Sotiris Missailidis Ygara da Silva Mendes Sheila Maria Barbosa de Lima Inactivated and Immunogenic SARS-CoV-2 for Safe Use in Immunoassays and as an Immunization Control for Non-Clinical Trials Viruses SARS-CoV-2 inactivation beta-propiolactone serial passages TCID<sub>50</sub> RT-qPCR non-clinical trials |
| title | Inactivated and Immunogenic SARS-CoV-2 for Safe Use in Immunoassays and as an Immunization Control for Non-Clinical Trials |
| title_full | Inactivated and Immunogenic SARS-CoV-2 for Safe Use in Immunoassays and as an Immunization Control for Non-Clinical Trials |
| title_fullStr | Inactivated and Immunogenic SARS-CoV-2 for Safe Use in Immunoassays and as an Immunization Control for Non-Clinical Trials |
| title_full_unstemmed | Inactivated and Immunogenic SARS-CoV-2 for Safe Use in Immunoassays and as an Immunization Control for Non-Clinical Trials |
| title_short | Inactivated and Immunogenic SARS-CoV-2 for Safe Use in Immunoassays and as an Immunization Control for Non-Clinical Trials |
| title_sort | inactivated and immunogenic sars cov 2 for safe use in immunoassays and as an immunization control for non clinical trials |
| topic | SARS-CoV-2 inactivation beta-propiolactone serial passages TCID<sub>50</sub> RT-qPCR non-clinical trials |
| url | https://www.mdpi.com/1999-4915/15/7/1486 |
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