Cargando…
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
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 re...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8688357/ https://www.ncbi.nlm.nih.gov/pubmed/34950112 http://dx.doi.org/10.3389/fmicb.2021.726231 |
_version_ | 1784618332423454720 |
---|---|
author | Kim, Dongbum Kim, Jinsoo Park, Sangkyu Kim, Minyoung Baek, Kyeongbin Kang, Mijeong Choi, Jun-Kyu Maharjan, Sony Akauliya, Madhav Lee, Younghee Kwon, Hyung-Joo |
author_facet | Kim, Dongbum Kim, Jinsoo Park, Sangkyu Kim, Minyoung Baek, Kyeongbin Kang, Mijeong Choi, Jun-Kyu Maharjan, Sony Akauliya, Madhav Lee, Younghee Kwon, Hyung-Joo |
author_sort | Kim, Dongbum |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8688357 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-86883572021-12-22 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 Kim, Dongbum Kim, Jinsoo Park, Sangkyu Kim, Minyoung Baek, Kyeongbin Kang, Mijeong Choi, Jun-Kyu Maharjan, Sony Akauliya, Madhav Lee, Younghee Kwon, Hyung-Joo Front Microbiol Microbiology 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. Frontiers Media S.A. 2021-12-07 /pmc/articles/PMC8688357/ /pubmed/34950112 http://dx.doi.org/10.3389/fmicb.2021.726231 Text en Copyright © 2021 Kim, Kim, Park, Kim, Baek, Kang, Choi, Maharjan, Akauliya, Lee and Kwon. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Microbiology Kim, Dongbum Kim, Jinsoo Park, Sangkyu Kim, Minyoung Baek, Kyeongbin Kang, Mijeong Choi, Jun-Kyu Maharjan, Sony Akauliya, Madhav Lee, Younghee Kwon, Hyung-Joo 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 |
title | 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 |
title_full | 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 |
title_fullStr | 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 |
title_full_unstemmed | 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 |
title_short | 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 |
title_sort | 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 |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8688357/ https://www.ncbi.nlm.nih.gov/pubmed/34950112 http://dx.doi.org/10.3389/fmicb.2021.726231 |
work_keys_str_mv | AT kimdongbum productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT kimjinsoo productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT parksangkyu productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT kimminyoung productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT baekkyeongbin productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT kangmijeong productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT choijunkyu productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT maharjansony productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT akauliyamadhav productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT leeyounghee productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein AT kwonhyungjoo productionofsarscov2nproteinspecificmonoclonalantibodyanditsapplicationinanelisabaseddetectionsystemandtargetingtheinteractionbetweenthespikecterminaldomainandnprotein |