tetano
Editor, Senior Moderator
Virol J
. 2022 Dec 10;19(1):213.
doi: 10.1186/s12985-022-01948-2.
Development of novel monoclonal antibodies against nsp12 of SARS-CoV-2
Mitsuhiro Machitani[SUP] 1 [/SUP], Junko Takei[SUP] 2 [/SUP], Mika K Kaneko[SUP] 2 [/SUP], Saori Ueki[SUP] 1 [/SUP], Hirofumi Ohashi[SUP] 3 4 [/SUP], Koichi Watashi[SUP] 3 4 [/SUP], Yukinari Kato[SUP] 2 5 [/SUP], Kenkichi Masutomi[SUP] 6 [/SUP]
Affiliations
Abstract
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.
Keywords: Monoclonal antibody; RNA-dependent RNA polymerase; SARS-CoV-2; nsp12.
. 2022 Dec 10;19(1):213.
doi: 10.1186/s12985-022-01948-2.
Development of novel monoclonal antibodies against nsp12 of SARS-CoV-2
Mitsuhiro Machitani[SUP] 1 [/SUP], Junko Takei[SUP] 2 [/SUP], Mika K Kaneko[SUP] 2 [/SUP], Saori Ueki[SUP] 1 [/SUP], Hirofumi Ohashi[SUP] 3 4 [/SUP], Koichi Watashi[SUP] 3 4 [/SUP], Yukinari Kato[SUP] 2 5 [/SUP], Kenkichi Masutomi[SUP] 6 [/SUP]
Affiliations
- PMID: 36496472
- DOI: 10.1186/s12985-022-01948-2
Abstract
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.
Keywords: Monoclonal antibody; RNA-dependent RNA polymerase; SARS-CoV-2; nsp12.