Inactivated and Immunogenic SARS-CoV-2 for Safe Use in Immunoassays and as an Immunization Control for Non-Clinical Trials

Author:

Gomes Mariana Pierre de Barros1,Linhares José Henrique Rezende1ORCID,dos Santos Tiago Pereira1,Pereira Renata Carvalho1,Santos Renata Tourinho1,da Silva Stephanie Almeida1,Souza Marta Cristina de Oliveira1,da Silva Juliana Fernandes Amorim1,Trindade Gisela Freitas1,Gomes Viviane Silva1,Barreto-Vieira Débora Ferreira2ORCID,Carvalho Milena Mouta Verdan França3,Ano Bom Ana Paula Dinis3,Gardinali Noemi Rovaris1ORCID,Müller Rodrigo4,Alves Nathalia dos Santos1ORCID,Moura Luma da Cruz1ORCID,Neves Patrícia Cristina da Costa3ORCID,Esteves Gabriela Santos5,Schwarcz Waleska Dias1,Missailidis Sotiris6,Mendes Ygara da Silva1ORCID,de Lima Sheila Maria Barbosa1ORCID

Affiliation:

1. Virological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, Brazil

2. Viral Morphology and Morphogenesis Laboratory, Oswaldo Cruz Institute/FIOCRUZ, Rio de Janeiro 21040-900, RJ, Brazil

3. Immunological Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, Brazil

4. Pre-Clinical Trials Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, Brazil

5. Recombinant Technology Laboratory, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, Brazil

6. Institute of Technology in Immunobiologicals, Bio-Manguinhos/FIOCRUZ, Rio de Janeiro 21040-900, RJ, Brazil

Abstract

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 AddaS03TM, 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.

Funder

Bio-Manguinhos/FIOCRUZ

Publisher

MDPI AG

Subject

Virology,Infectious Diseases

Reference52 articles.

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4. OMS (2023, June 23). Declara Fim Da Emergência de Saúde Pública de Importância Internacional Referente à COVID-19—OPAS/OMS | Organização Pan-Americana Da Saúde. Available online: https://www.paho.org/pt/noticias/5-5-2023-oms-declara-fim-da-emergencia-saude-publica-importancia-internacional-referente.

5. Mathieu, E., Ritchie, H., Rodés-Guirao, L., Appel, C., Giattino, C., Hasell, J., Macdonald, B., Dattani, S., Beltekian, D., and Ortiz-Ospina, E. (2023, June 23). Coronavirus Pandemic (COVID-19). Available online: https://ourworldindata.org/covid-vaccinations.

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