Development of novel monoclonal antibodies against nsp12 of SARS-CoV-2

Virology Journal

Brief Report

Mika K. Kaneko¹  Junko Takei¹  Yukinari Kato²  Kenkichi Masutomi³  Saori Ueki³  Mitsuhiro Machitani³  Koichi Watashi⁴  Hirofumi Ohashi⁴ 

[1] Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-Machi, Aoba-Ku, 980-8575, Sendai, Miyagi, Japan;Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-Machi, Aoba-Ku, 980-8575, Sendai, Miyagi, Japan;Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-Machi, Aoba-Ku, 980-8575, Sendai, Miyagi, Japan;Division of Cancer Stem Cell, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-Ku, 104-0045, Tokyo, Japan;Research Center for Drug and Vaccine Development, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku, 162-8640, Tokyo, Japan;Department of Virology II, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku, 162-8640, Tokyo, Japan;

关键词: SARS-CoV-2;    RNA-dependent RNA polymerase;    nsp12;    Monoclonal antibody;   

DOI  :  10.1186/s12985-022-01948-2

 received in 2022-03-25, accepted in 2022-12-06,  发布年份 2022

来源: Springer

【 摘 要 】

A novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has caused a global pandemic of coronavirus disease 19. Coronaviruses, including SARS-CoV-2, use RNA-dependent RNA polymerase (RdRP) for viral replication and transcription. Since RdRP is a promising therapeutic target for infection of SARS-CoV-2, it would be beneficial to develop new experimental tools for analysis of the RdRP reaction of SARS-CoV-2. Here, we succeeded to develop novel mouse monoclonal antibodies (mAbs) that recognize SARS-CoV-2 nsp12, catalytic subunit of the RdRP. These anti-nsp12 mAbs, RdMab-2, -13, and -20, specifically recognize SARS-CoV-2 nsp12 by western blotting analysis, while they exhibit less or no cross-reactivity to SARS-CoV nsp12. In addition, SARS-CoV-2 nsp12 was successfully immunoprecipitated using RdMab-2 from lysates of cells overexpressing SARS-CoV-2 nsp12. RdMab-2 was able to detect SARS-CoV-2 nsp12 transiently expressed in established culture cells such as HEK293T cells by indirect immunofluorescence technique. These novel mAbs against SARS-CoV-2 nsp12 are useful to elucidate the RdRP reaction of SARS-CoV-2 and biological cell response against it.

【 授权许可】

CC BY   
© The Author(s) 2022

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