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Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex

Middle East respiratory syndrome coronavirus (MERS-CoV) uses the spike (S) glycoprotein to recognize and enter target cells. In this study, we selected two epitope peptide sequences within the receptor binding domain (RBD) of the MERS-CoV S protein. We used a complex consisting of the epitope peptid...

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Autores principales: Park, Byoung Kwon, Maharjan, Sony, Lee, Su In, Kim, Jinsoo, Bae, Joon-Yong, Park, Man-Seong, Kwon, Hyung-Joo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society for Biochemistry and Molecular Biology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6605520/
https://www.ncbi.nlm.nih.gov/pubmed/30355437
http://dx.doi.org/10.5483/BMBRep.2019.52.6.185
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author Park, Byoung Kwon
Maharjan, Sony
Lee, Su In
Kim, Jinsoo
Bae, Joon-Yong
Park, Man-Seong
Kwon, Hyung-Joo
author_facet Park, Byoung Kwon
Maharjan, Sony
Lee, Su In
Kim, Jinsoo
Bae, Joon-Yong
Park, Man-Seong
Kwon, Hyung-Joo
author_sort Park, Byoung Kwon
collection PubMed
description Middle East respiratory syndrome coronavirus (MERS-CoV) uses the spike (S) glycoprotein to recognize and enter target cells. In this study, we selected two epitope peptide sequences within the receptor binding domain (RBD) of the MERS-CoV S protein. We used a complex consisting of the epitope peptide of the MERS-CoV S protein and CpG-DNA encapsulated in liposome complex to immunize mice, and produced the monoclonal antibodies 506-2G10G5 and 492-1G10E4E2. The western blotting data showed that both monoclonal antibodies detected the S protein and immunoprecipitated the native form of the S protein. Indirect immunofluorescence and confocal analysis suggested strong reactivity of the antibodies towards the S protein of MERS-CoV virus infected Vero cells. Furthermore, the 506-2G10G5 monoclonal antibody significantly reduced plaque formation in MERS-CoV infected Vero cells compared to normal mouse IgG and 492-1G10E4E2. Thus, we successfully produced a monoclonal antibody directed against the RBD domain of the S protein which could be used in the development of diagnostics and therapeutic applications in the future.
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spelling pubmed-66055202019-07-10 Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex Park, Byoung Kwon Maharjan, Sony Lee, Su In Kim, Jinsoo Bae, Joon-Yong Park, Man-Seong Kwon, Hyung-Joo BMB Rep Articles Middle East respiratory syndrome coronavirus (MERS-CoV) uses the spike (S) glycoprotein to recognize and enter target cells. In this study, we selected two epitope peptide sequences within the receptor binding domain (RBD) of the MERS-CoV S protein. We used a complex consisting of the epitope peptide of the MERS-CoV S protein and CpG-DNA encapsulated in liposome complex to immunize mice, and produced the monoclonal antibodies 506-2G10G5 and 492-1G10E4E2. The western blotting data showed that both monoclonal antibodies detected the S protein and immunoprecipitated the native form of the S protein. Indirect immunofluorescence and confocal analysis suggested strong reactivity of the antibodies towards the S protein of MERS-CoV virus infected Vero cells. Furthermore, the 506-2G10G5 monoclonal antibody significantly reduced plaque formation in MERS-CoV infected Vero cells compared to normal mouse IgG and 492-1G10E4E2. Thus, we successfully produced a monoclonal antibody directed against the RBD domain of the S protein which could be used in the development of diagnostics and therapeutic applications in the future. Korean Society for Biochemistry and Molecular Biology 2019-06 2019-06-30 /pmc/articles/PMC6605520/ /pubmed/30355437 http://dx.doi.org/10.5483/BMBRep.2019.52.6.185 Text en Copyright © 2019 by the The Korean Society for Biochemistry and Molecular Biology This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Articles
Park, Byoung Kwon
Maharjan, Sony
Lee, Su In
Kim, Jinsoo
Bae, Joon-Yong
Park, Man-Seong
Kwon, Hyung-Joo
Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex
title Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex
title_full Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex
title_fullStr Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex
title_full_unstemmed Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex
title_short Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex
title_sort generation and characterization of a monoclonal antibody against mers-cov targeting the spike protein using a synthetic peptide epitope-cpg-dna-liposome complex
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6605520/
https://www.ncbi.nlm.nih.gov/pubmed/30355437
http://dx.doi.org/10.5483/BMBRep.2019.52.6.185
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