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.
【저자키워드】 SARS-CoV-2, ELISA, Spike protein, virus detection, N protein, bait and prey, anti-SARS-CoV-2 N antibody, 【초록키워드】 coronavirus, spike, antibody, SARS-COV-2 infection, monoclonal antibody, Proteins, MERS-CoV, Spread, sensitivity, clade, fusion protein, SARS-CoV-2 infections, diagnose, Interaction, C-terminal domain, culture media, this system, Vero cells, human-to-human transmission, infected cell, Vero Cell, produced, detect, reported, recognize, the spike protein, interact, the N protein, cell lysate, pfu, the SARS-CoV-2, 【제목키워드】 Production, detection, application, SARS-CoV-2 N, targeting, System, the Spike,