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Front Microbiol. 2021 Dec 07;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.

Frontiers in microbiology

Dongbum Kim, Jinsoo Kim, Sangkyu Park, Minyoung Kim, Kyeongbin Baek, Mijeong Kang, Jun-Kyu Choi, Sony Maharjan, Madhav Akauliya, Younghee Lee, Hyung-Joo Kwon

Affiliations

  1. Institute of Medical Science, College of Medicine, Hallym University, Chuncheon, South Korea.
  2. Department of Microbiology, College of Medicine, Hallym University, Chuncheon, South Korea.
  3. Department of Biochemistry, College of Natural Sciences, Chungbuk National University, Cheongju, South Korea.

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.

Copyright © 2021 Kim, Kim, Park, Kim, Baek, Kang, Choi, Maharjan, Akauliya, Lee and Kwon.

Keywords: ELISA; N protein; SARS-CoV-2; anti-SARS-CoV-2 N antibody; bait and prey; spike protein; virus detection

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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