3D8, a nucleic acid-hydrolyzing scFv, confers antiviral activity against SARS-CoV-2 and multiple coronaviruses in vitro

Lee, G.; Budhathoki, S.; Choi, H. S.; Oh, K.-j.; Lee, G.-Y.; Ham, Y. K.; Kim, Y. J.; Lim, Y. R.; Hoang, P. T.; Lee, Y.; Lim, S.-W.; Kim, J.-M.; Cho, S.; Song, J.-W.; Lee, S.; Kim, W.-K.

doi:10.1101/2020.11.25.398909

bioRxiv · 10.1101/2020.11.25.398909

3D8, a nucleic acid-hydrolyzing scFv, confers antiviral activity against SARS-CoV-2 and multiple coronaviruses in vitro

Lee, G.·Budhathoki, S.·Choi, H. S.·Oh, K.-j.·Lee, G.-Y.·Ham, Y. K.·Kim, Y. J.·Lim, Y. R.·Hoang, P. T.·Lee, Y.·Lim, S.-W.·Kim, J.-M.·Cho, S.·Song, J.-W.·Lee, S.·Kim, W.-K.

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Abstract

BackgroundThe current pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the etiology of Coronavirus-induced disease 19 (COVID-19) and poses a critical public health threat worldwide. Effective therapeutics and vaccines against multiple coronaviruses remain unavailable. Single chain variable fragment (scFv), a recombinant antibody exhibits broad-spectrum antiviral activity against DNA and RNA viruses owing to its nucleic acid-hydrolyzing property. ObjectiveThis study is aimed to investigate an antiviral activity of 3D8 scFv against SARS-CoV-2 and other coronaviruses. Methods3D8, a recombinant scFv antibody was evaluated for antiviral activity against SARS-CoV-2, HCoV-OC43 and PEDV in Vero E6 cell cultures. Viral growth was quantified with quantitative RT-qPCR and plaque assay. Nucleic acid hydrolyzing activity of 3D8 was assessed through abzyme assays of in vitro viral transcripts and cell viability was determined by MTT assay. Results3D8 inhibited the replication of SARS-CoV-2, human coronavirus OC43 (HCoV-OC43), and porcine epidemic diarrhea virus (PEDV). Our results revealed the prophylactic and therapeutic effects of 3D8 scFv against SARS-CoV-2 in Vero E6 cells. Immunoblot and plaque assays showed the reduction of coronavirus nucleoproteins and infectious particles respectively in 3D8 scFv-treated cells. ConclusionsThis data demonstrates the broad-spectrum antiviral activity of 3D8 against SARS-CoV-2 and other coronaviruses. Thus, it could be considered a potential antiviral countermeasure against SARS-CoV-2 and zoonotic coronaviruses.

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Lee, G., Budhathoki, S., Choi, H. S., Oh, K.-j., Lee, G.-Y., Ham, Y. K., Kim, Y. J., Lim, Y. R., Hoang, P. T., Lee, Y., Lim, S.-W., Kim, J.-M., Cho, S., Song, J.-W., Lee, S., Kim, W.-K.. 2020-11-25. 3D8, a nucleic acid-hydrolyzing scFv, confers antiviral activity against SARS-CoV-2 and multiple coronaviruses in vitro. https://doi.org/10.1101/2020.11.25.398909

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