Broad-spectrum antiviral activity of 3D8, a nucleic acid-hydrolyzing single chain variable fragment (scFv), targeting SARS-CoV-2 and multiple coronaviruses in vitro

• Published in: bioRxiv
• Main Authors: Lee, Gunsup, Budhathoki, Shailesh, Geum-Young, Lee, Kwang-Ji Oh, Ham, Yeon Kyoung, Young Jun Kim, Lim, Ye Rin, Phuong Thi Hoang, Lee, Yongjun, Seok-Won Lim, Jun-Mo, Kim, Cho, Seungchan, Tai-Hyun, Kim, Jin-Won, Song, Lee, Sukchan, Won-Keun, Kim
• Format: Paper
• Language: English
• Published: Cold Spring Harbor Cold Spring Harbor Laboratory Press 15.02.2021
• Subjects: Antiviral activity; Cell viability; Coronaviridae; Coronaviruses; COVID-19; Diarrhea; DNA viruses; Etiology; Nucleoproteins; Pandemics; Plaque assay; Public health; RNA viruses; Severe acute respiratory syndrome coronavirus 2; Transmissible gastroenteritis
• DOI: 10.1101/2020.11.25.398909

Abstract Background The 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. Objective This study is aimed to investigate an antiviral activity of 3D8 scFv against SARS-CoV-2 and other coronaviruses. Methods 3D8, 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. Results 3D8 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. Conclusions This 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. Competing Interest Statement The authors have declared no competing interest. Footnotes * Title changed; Main context significantly revised; Figure 1 added; All of figures updated; Supplemental tables updated.