tetano
Editor, Senior Moderator
Front Microbiol
. 2021 Dec 7;12:726231.
doi: 10.3389/fmicb.2021.726231. eCollection 2021.
Production of SARS-CoV-2 N Protein-Specific Monoclonal Antibody and Its Application in an ELISA-Based Detection System and Targeting the Interaction Between the Spike C-Terminal Domain and N Protein
Dongbum Kim[SUP] 1 [/SUP], Jinsoo Kim[SUP] 2 [/SUP], Sangkyu Park[SUP] 3 [/SUP], Minyoung Kim[SUP] 2 [/SUP], Kyeongbin Baek[SUP] 2 [/SUP], Mijeong Kang[SUP] 2 [/SUP], Jun-Kyu Choi[SUP] 3 [/SUP], Sony Maharjan[SUP] 1 [/SUP], Madhav Akauliya[SUP] 2 [/SUP], Younghee Lee[SUP] 3 [/SUP], Hyung-Joo Kwon[SUP] 1 2 [/SUP]
Affiliations
Abstract
SARS-CoV-2 infections continue to spread quickly by human-to-human transmission around the world. Therefore, developing methods to rapidly detect SARS-CoV-2 with high sensitivity are still urgently needed. We produced a monoclonal antibody that specifically detects the N protein of SARS-CoV-2 and recognizes N protein in cell lysates of SARS-CoV-2-infected Vero cells but not in cell lysates of MERS-CoV- or HCoV-OC43-infected Vero cells. This antibody recognized N protein in SARS-CoV-2 clades S, GR, and GH and recognized N protein in all the SARS-CoV-2 clades to ∼300 pfu. Previously, we reported that the coronavirus N protein interacts with the C-terminal domain of the spike protein (Spike CD). In this study, we developed an ELISA-based "bait and prey" system to confirm the interaction between SARS-CoV-2 Spike CD and N protein using recombinant fusion proteins. Furthermore, this system can be modified to quantitatively detect SARS-CoV-2 in culture media of infected cells by monitoring the interaction between the recombinant Spike CD fusion protein and the viral N protein, which is captured by the N protein-specific antibody. Therefore, we conclude that our N protein-specific monoclonal antibody and our ELISA-based bait and prey system could be used to diagnose SARS-CoV-2 infections.
Keywords: ELISA; N protein; SARS-CoV-2; anti-SARS-CoV-2 N antibody; bait and prey; spike protein; virus detection.
. 2021 Dec 7;12:726231.
doi: 10.3389/fmicb.2021.726231. eCollection 2021.
Production of SARS-CoV-2 N Protein-Specific Monoclonal Antibody and Its Application in an ELISA-Based Detection System and Targeting the Interaction Between the Spike C-Terminal Domain and N Protein
Dongbum Kim[SUP] 1 [/SUP], Jinsoo Kim[SUP] 2 [/SUP], Sangkyu Park[SUP] 3 [/SUP], Minyoung Kim[SUP] 2 [/SUP], Kyeongbin Baek[SUP] 2 [/SUP], Mijeong Kang[SUP] 2 [/SUP], Jun-Kyu Choi[SUP] 3 [/SUP], Sony Maharjan[SUP] 1 [/SUP], Madhav Akauliya[SUP] 2 [/SUP], Younghee Lee[SUP] 3 [/SUP], Hyung-Joo Kwon[SUP] 1 2 [/SUP]
Affiliations
- PMID: 34950112
- PMCID: PMC8688357
- DOI: 10.3389/fmicb.2021.726231
Abstract
SARS-CoV-2 infections continue to spread quickly by human-to-human transmission around the world. Therefore, developing methods to rapidly detect SARS-CoV-2 with high sensitivity are still urgently needed. We produced a monoclonal antibody that specifically detects the N protein of SARS-CoV-2 and recognizes N protein in cell lysates of SARS-CoV-2-infected Vero cells but not in cell lysates of MERS-CoV- or HCoV-OC43-infected Vero cells. This antibody recognized N protein in SARS-CoV-2 clades S, GR, and GH and recognized N protein in all the SARS-CoV-2 clades to ∼300 pfu. Previously, we reported that the coronavirus N protein interacts with the C-terminal domain of the spike protein (Spike CD). In this study, we developed an ELISA-based "bait and prey" system to confirm the interaction between SARS-CoV-2 Spike CD and N protein using recombinant fusion proteins. Furthermore, this system can be modified to quantitatively detect SARS-CoV-2 in culture media of infected cells by monitoring the interaction between the recombinant Spike CD fusion protein and the viral N protein, which is captured by the N protein-specific antibody. Therefore, we conclude that our N protein-specific monoclonal antibody and our ELISA-based bait and prey system could be used to diagnose SARS-CoV-2 infections.
Keywords: ELISA; N protein; SARS-CoV-2; anti-SARS-CoV-2 N antibody; bait and prey; spike protein; virus detection.