Testing the Efficacy of COVID-19 Vaccine and Therapeutic Candidates Using Vero 76 Cell Line = Testing the Efficacy of COVID-19 Vaccine and Therapeutic Candidates Using Vero 76 Cell Line

다국어 초록 (Multilingual Abstract)

Background
COVID-19 is a respiratory infectious disease that first emerged in Wuhan, China in December 2019 and has spread worldwide. It is necessary to develop preventive vaccines and therapeutics for overcoming COVID-19. For this purpose, , in vitro efficacy testing of the applicants is essential in the early stages of development. We want to share our experience of in vitro efficacy testing Methods using SARS-CoV-2 through this presentation.
Methods
All tests using SARS-CoV-2 were performed within the Biosafely Level 3 (BL3) facility of Masan National Tuberculosis Hospital (MNTH). And the protocols were approved by the Institutional Biosafety Committee (IBC) of MNTH prior to the study. Vaccine candidate neutralizing Serums were obtained by collecting blood from animals immunized with vaccine candidates. In 6 well plates, the Vero 76 cells were pre-culture to sub-confluence. The serum dilution and the virus was mixed in a 1:1 volume and neutralized. The serumvirus mixture was added to the cells for infection and the uninfected virus was removed. Add media containing 1% agarose, and solidify completely. After culturing for 3 days, fix with formaldehyde, remove agarose, and stain with crystal violet. The neutralizing effect was calculated by counting the plaques and comparing them with the control. Therapeutic candidates Infect SARS-CoV-2 to Vero 76 cells with an MOI 0.01. Add the test substance prepared for each concentration and Remdesivir, a positive control, and incubate. The culture medium and cell lysate are collected, the therapeutic effect is confirmed by comparing Results with the control group by Methods such as plaque assay, qPCR and WB.
Results
We established our optimized in vitro protocol for testing the efficacy of vaccines and therapeutic candidates against COVID-19 quantitatively.
Conclusion
The established Methods would be useful in confirming the efficacy of COVID-19 vaccine candidates and therapeutic candidates in the preclinical stage.