Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex
Identifieur interne : 000302 ( Pmc/Checkpoint ); précédent : 000301; suivant : 000303Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex
Auteurs : Byoung Kwon Park [Corée du Sud] ; Sony Maharjan [Corée du Sud] ; Su In Lee [Corée du Sud] ; Jinsoo Kim [Corée du Sud] ; Joon-Yong Bae [Corée du Sud] ; Man-Seong Park [Corée du Sud] ; Hyung-Joo Kwon [Corée du Sud]Source :
- BMB Reports [ 1976-6696 ] ; 2019.
Abstract
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.
Url:
DOI: 10.5483/BMBRep.2019.52.6.185
PubMed: 30355437
PubMed Central: 6605520
Affiliations:
Links toward previous steps (curation, corpus...)
Links to Exploration step
PMC:6605520Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex</title>
<author><name sortKey="Park, Byoung Kwon" sort="Park, Byoung Kwon" uniqKey="Park B" first="Byoung Kwon" last="Park">Byoung Kwon Park</name>
<affiliation wicri:level="1"><nlm:aff id="af1-bmb-52-397">Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Maharjan, Sony" sort="Maharjan, Sony" uniqKey="Maharjan S" first="Sony" last="Maharjan">Sony Maharjan</name>
<affiliation wicri:level="1"><nlm:aff id="af1-bmb-52-397">Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Lee, Su In" sort="Lee, Su In" uniqKey="Lee S" first="Su In" last="Lee">Su In Lee</name>
<affiliation wicri:level="1"><nlm:aff id="af1-bmb-52-397">Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Kim, Jinsoo" sort="Kim, Jinsoo" uniqKey="Kim J" first="Jinsoo" last="Kim">Jinsoo Kim</name>
<affiliation wicri:level="1"><nlm:aff id="af2-bmb-52-397">Department of Microbiology, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Bae, Joon Yong" sort="Bae, Joon Yong" uniqKey="Bae J" first="Joon-Yong" last="Bae">Joon-Yong Bae</name>
<affiliation wicri:level="1"><nlm:aff id="af3-bmb-52-397">Department of Microbiology, College of Medicine, and the Institute for Viral Diseases, Korea University, Seoul 02841,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Park, Man Seong" sort="Park, Man Seong" uniqKey="Park M" first="Man-Seong" last="Park">Man-Seong Park</name>
<affiliation wicri:level="1"><nlm:aff id="af3-bmb-52-397">Department of Microbiology, College of Medicine, and the Institute for Viral Diseases, Korea University, Seoul 02841,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Kwon, Hyung Joo" sort="Kwon, Hyung Joo" uniqKey="Kwon H" first="Hyung-Joo" last="Kwon">Hyung-Joo Kwon</name>
<affiliation wicri:level="1"><nlm:aff id="af1-bmb-52-397">Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="af2-bmb-52-397">Department of Microbiology, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">30355437</idno>
<idno type="pmc">6605520</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6605520</idno>
<idno type="RBID">PMC:6605520</idno>
<idno type="doi">10.5483/BMBRep.2019.52.6.185</idno>
<date when="2019">2019</date>
<idno type="wicri:Area/Pmc/Corpus">000A84</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000A84</idno>
<idno type="wicri:Area/Pmc/Curation">000A84</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000A84</idno>
<idno type="wicri:Area/Pmc/Checkpoint">000302</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000302</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex</title>
<author><name sortKey="Park, Byoung Kwon" sort="Park, Byoung Kwon" uniqKey="Park B" first="Byoung Kwon" last="Park">Byoung Kwon Park</name>
<affiliation wicri:level="1"><nlm:aff id="af1-bmb-52-397">Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Maharjan, Sony" sort="Maharjan, Sony" uniqKey="Maharjan S" first="Sony" last="Maharjan">Sony Maharjan</name>
<affiliation wicri:level="1"><nlm:aff id="af1-bmb-52-397">Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Lee, Su In" sort="Lee, Su In" uniqKey="Lee S" first="Su In" last="Lee">Su In Lee</name>
<affiliation wicri:level="1"><nlm:aff id="af1-bmb-52-397">Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Kim, Jinsoo" sort="Kim, Jinsoo" uniqKey="Kim J" first="Jinsoo" last="Kim">Jinsoo Kim</name>
<affiliation wicri:level="1"><nlm:aff id="af2-bmb-52-397">Department of Microbiology, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Bae, Joon Yong" sort="Bae, Joon Yong" uniqKey="Bae J" first="Joon-Yong" last="Bae">Joon-Yong Bae</name>
<affiliation wicri:level="1"><nlm:aff id="af3-bmb-52-397">Department of Microbiology, College of Medicine, and the Institute for Viral Diseases, Korea University, Seoul 02841,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Park, Man Seong" sort="Park, Man Seong" uniqKey="Park M" first="Man-Seong" last="Park">Man-Seong Park</name>
<affiliation wicri:level="1"><nlm:aff id="af3-bmb-52-397">Department of Microbiology, College of Medicine, and the Institute for Viral Diseases, Korea University, Seoul 02841,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Kwon, Hyung Joo" sort="Kwon, Hyung Joo" uniqKey="Kwon H" first="Hyung-Joo" last="Kwon">Hyung-Joo Kwon</name>
<affiliation wicri:level="1"><nlm:aff id="af1-bmb-52-397">Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="af2-bmb-52-397">Department of Microbiology, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</nlm:aff>
<country xml:lang="fr">Corée du Sud</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
</analytic>
<series><title level="j">BMB Reports</title>
<idno type="ISSN">1976-6696</idno>
<idno type="eISSN">1976-670X</idno>
<imprint><date when="2019">2019</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p>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.</p>
</div>
</front>
<back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Zumla, A" uniqKey="Zumla A">A Zumla</name>
</author>
<author><name sortKey="Chan, Jf" uniqKey="Chan J">JF Chan</name>
</author>
<author><name sortKey="Azhar, Ei" uniqKey="Azhar E">EI Azhar</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Du, L" uniqKey="Du L">L Du</name>
</author>
<author><name sortKey="Yang, Y" uniqKey="Yang Y">Y Yang</name>
</author>
<author><name sortKey="Zhou, Y" uniqKey="Zhou Y">Y Zhou</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Mackay, Im" uniqKey="Mackay I">IM Mackay</name>
</author>
<author><name sortKey="Arden, Ke" uniqKey="Arden K">KE Arden</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Zaki, Am" uniqKey="Zaki A">AM Zaki</name>
</author>
<author><name sortKey="Van Boheemen, S" uniqKey="Van Boheemen S">S van Boheemen</name>
</author>
<author><name sortKey="Bestebroer, Tm" uniqKey="Bestebroer T">TM Bestebroer</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Omrani, As" uniqKey="Omrani A">AS Omrani</name>
</author>
<author><name sortKey="Al Tawfiq, Ja" uniqKey="Al Tawfiq J">JA Al-Tawfiq</name>
</author>
<author><name sortKey="Memish, Za" uniqKey="Memish Z">ZA Memish</name>
</author>
</analytic>
</biblStruct>
<biblStruct></biblStruct>
<biblStruct><analytic><author><name sortKey="Ki, M" uniqKey="Ki M">M Ki</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Durai, P" uniqKey="Durai P">P Durai</name>
</author>
<author><name sortKey="Batool, M" uniqKey="Batool M">M Batool</name>
</author>
<author><name sortKey="Shah, M" uniqKey="Shah M">M Shah</name>
</author>
<author><name sortKey="Choi, S" uniqKey="Choi S">S Choi</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Lambour, J" uniqKey="Lambour J">J Lambour</name>
</author>
<author><name sortKey="Naranjo Gomez, M" uniqKey="Naranjo Gomez M">M Naranjo-Gomez</name>
</author>
<author><name sortKey="Piechaczyk, M" uniqKey="Piechaczyk M">M Piechaczyk</name>
</author>
<author><name sortKey="Pelegrin, M" uniqKey="Pelegrin M">M Pelegrin</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Scott, Am" uniqKey="Scott A">AM Scott</name>
</author>
<author><name sortKey="Allison, Jp" uniqKey="Allison J">JP Allison</name>
</author>
<author><name sortKey="Wolchok, Jd" uniqKey="Wolchok J">JD Wolchok</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Wang, Q" uniqKey="Wang Q">Q Wang</name>
</author>
<author><name sortKey="Wong, G" uniqKey="Wong G">G Wong</name>
</author>
<author><name sortKey="Lu, G" uniqKey="Lu G">G Lu</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Gao, J" uniqKey="Gao J">J Gao</name>
</author>
<author><name sortKey="Lu, G" uniqKey="Lu G">G Lu</name>
</author>
<author><name sortKey="Qi, J" uniqKey="Qi J">J Qi</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Wang, N" uniqKey="Wang N">N Wang</name>
</author>
<author><name sortKey="Shi, X" uniqKey="Shi X">X Shi</name>
</author>
<author><name sortKey="Jiang, L" uniqKey="Jiang L">L Jiang</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Raj, Vs" uniqKey="Raj V">VS Raj</name>
</author>
<author><name sortKey="Mou, H" uniqKey="Mou H">H Mou</name>
</author>
<author><name sortKey="Smits, Sl" uniqKey="Smits S">SL Smits</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Kim, D" uniqKey="Kim D">D Kim</name>
</author>
<author><name sortKey="Kwon, Hj" uniqKey="Kwon H">HJ Kwon</name>
</author>
<author><name sortKey="Lee, Y" uniqKey="Lee Y">Y Lee</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Rhee, Jw" uniqKey="Rhee J">JW Rhee</name>
</author>
<author><name sortKey="Kim, D" uniqKey="Kim D">D Kim</name>
</author>
<author><name sortKey="Park" uniqKey="Park">Park</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Park, Bk" uniqKey="Park B">BK Park</name>
</author>
<author><name sortKey="Choi, Sh" uniqKey="Choi S">SH Choi</name>
</author>
<author><name sortKey="Kim, Ye" uniqKey="Kim Y">YE Kim</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Park, Bk" uniqKey="Park B">BK Park</name>
</author>
<author><name sortKey="Lee, Si" uniqKey="Lee S">SI Lee</name>
</author>
<author><name sortKey="Bae, Jy" uniqKey="Bae J">JY Bae</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Salazar, G" uniqKey="Salazar G">G Salazar</name>
</author>
<author><name sortKey="Zhang, N" uniqKey="Zhang N">N Zhang</name>
</author>
<author><name sortKey="Fu, Tm" uniqKey="Fu T">TM Fu</name>
</author>
<author><name sortKey="An, Z" uniqKey="An Z">Z An</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Park, Bk" uniqKey="Park B">BK Park</name>
</author>
<author><name sortKey="Park, Jy" uniqKey="Park J">JY Park</name>
</author>
<author><name sortKey="Kim, Th" uniqKey="Kim T">TH Kim</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Wu, G" uniqKey="Wu G">G Wu</name>
</author>
<author><name sortKey="Maharjan, S" uniqKey="Maharjan S">S Maharjan</name>
</author>
<author><name sortKey="Kim, D" uniqKey="Kim D">D Kim</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Park, Bk" uniqKey="Park B">BK Park</name>
</author>
<author><name sortKey="Gautam, A" uniqKey="Gautam A">A Gautam</name>
</author>
<author><name sortKey="Maharjan, S" uniqKey="Maharjan S">S Maharjan</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Wu, G" uniqKey="Wu G">G Wu</name>
</author>
<author><name sortKey="Kim, D" uniqKey="Kim D">D Kim</name>
</author>
<author><name sortKey="Kim, Jn" uniqKey="Kim J">JN Kim</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Kim, D" uniqKey="Kim D">D Kim</name>
</author>
<author><name sortKey="Kwon, S" uniqKey="Kwon S">S Kwon</name>
</author>
<author><name sortKey="Rhee, Jw" uniqKey="Rhee J">JW Rhee</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Yokoyama, Wm" uniqKey="Yokoyama W">WM Yokoyama</name>
</author>
<author><name sortKey="Christensen, M" uniqKey="Christensen M">M Christensen</name>
</author>
<author><name sortKey="Santos, Gd" uniqKey="Santos G">GD Santos</name>
</author>
<author><name sortKey="Miller, D" uniqKey="Miller D">D Miller</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Maharjan, S" uniqKey="Maharjan S">S Maharjan</name>
</author>
<author><name sortKey="Park, Bk" uniqKey="Park B">BK Park</name>
</author>
<author><name sortKey="Lee, Si" uniqKey="Lee S">SI Lee</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Maharjan, S" uniqKey="Maharjan S">S Maharjan</name>
</author>
<author><name sortKey="Park, Bk" uniqKey="Park B">BK Park</name>
</author>
<author><name sortKey="Lee, Si" uniqKey="Lee S">SI Lee</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Jo, Hs" uniqKey="Jo H">HS Jo</name>
</author>
<author><name sortKey="Eum, Ws" uniqKey="Eum W">WS Eum</name>
</author>
<author><name sortKey="Park, Ey" uniqKey="Park E">EY Park</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Lee, Ew" uniqKey="Lee E">EW Lee</name>
</author>
<author><name sortKey="Oh, W" uniqKey="Oh W">W Oh</name>
</author>
<author><name sortKey="Song, Hp" uniqKey="Song H">HP Song</name>
</author>
<author><name sortKey="Kim, Wk" uniqKey="Kim W">WK Kim</name>
</author>
</analytic>
</biblStruct>
</listBibl>
</div1>
</back>
</TEI>
<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">BMB Rep</journal-id>
<journal-id journal-id-type="iso-abbrev">BMB Rep</journal-id>
<journal-title-group><journal-title>BMB Reports</journal-title>
</journal-title-group>
<issn pub-type="ppub">1976-6696</issn>
<issn pub-type="epub">1976-670X</issn>
<publisher><publisher-name>Korean Society for Biochemistry and Molecular Biology</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">30355437</article-id>
<article-id pub-id-type="pmc">6605520</article-id>
<article-id pub-id-type="doi">10.5483/BMBRep.2019.52.6.185</article-id>
<article-id pub-id-type="publisher-id">bmb-52-397</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Articles</subject>
</subj-group>
</article-categories>
<title-group><article-title>Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Park</surname>
<given-names>Byoung Kwon</given-names>
</name>
<xref ref-type="aff" rid="af1-bmb-52-397">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Maharjan</surname>
<given-names>Sony</given-names>
</name>
<xref ref-type="aff" rid="af1-bmb-52-397">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Lee</surname>
<given-names>Su In</given-names>
</name>
<xref ref-type="aff" rid="af1-bmb-52-397">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Kim</surname>
<given-names>Jinsoo</given-names>
</name>
<xref ref-type="aff" rid="af2-bmb-52-397">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Bae</surname>
<given-names>Joon-Yong</given-names>
</name>
<xref ref-type="aff" rid="af3-bmb-52-397">3</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Park</surname>
<given-names>Man-Seong</given-names>
</name>
<xref ref-type="aff" rid="af3-bmb-52-397">3</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Kwon</surname>
<given-names>Hyung-Joo</given-names>
</name>
<xref ref-type="aff" rid="af1-bmb-52-397">1</xref>
<xref ref-type="aff" rid="af2-bmb-52-397">2</xref>
<xref rid="c1-bmb-52-397" ref-type="corresp">*</xref>
</contrib>
</contrib-group>
<aff id="af1-bmb-52-397"><label>1</label>
Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</aff>
<aff id="af2-bmb-52-397"><label>2</label>
Department of Microbiology, College of Medicine, Hallym University, Chuncheon 24252,<country>Korea</country>
</aff>
<aff id="af3-bmb-52-397"><label>3</label>
Department of Microbiology, College of Medicine, and the Institute for Viral Diseases, Korea University, Seoul 02841,<country>Korea</country>
</aff>
<author-notes><corresp id="c1-bmb-52-397"><label>*</label>
Corresponding author. Tel: +82-33-248-2635; Fax: +82-33-241-3640; E-mail: <email>hjookwon@hallym.ac.kr</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub"><month>6</month>
<year>2019</year>
</pub-date>
<pub-date pub-type="epub"><day>30</day>
<month>6</month>
<year>2019</year>
</pub-date>
<volume>52</volume>
<issue>6</issue>
<fpage>397</fpage>
<lpage>402</lpage>
<history><date date-type="received"><day>08</day>
<month>8</month>
<year>2018</year>
</date>
<date date-type="rev-recd"><day>05</day>
<month>9</month>
<year>2018</year>
</date>
<date date-type="accepted"><day>11</day>
<month>10</month>
<year>2018</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2019 by the The Korean Society for Biochemistry and Molecular Biology</copyright-statement>
<copyright-year>2019</copyright-year>
<license license-type="open-access"><license-p>This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by-nc/4.0">http://creativecommons.org/licenses/by-nc/4.0</ext-link>
) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
</license>
</permissions>
<abstract><p>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.</p>
</abstract>
<kwd-group><kwd>B cell epitope</kwd>
<kwd>Lipoplex (O)</kwd>
<kwd>MERS-CoV</kwd>
<kwd>Monoclonal antibody</kwd>
<kwd>Spike protein</kwd>
</kwd-group>
</article-meta>
</front>
<floats-group><fig id="f1-bmb-52-397" orientation="portrait" position="float"><label>Fig. 1</label>
<caption><p>Production of the B cell epitope-specific antibody. (A) Schematic representation of the MERS-CoV S protein. The S protein consists of S1 and S2 subunits. RBD, receptor binding domain; FP, fusion peptide; HR1 and HR2, heptad repeat region 1 and 2; TM, transmembrane; CP, cytoplasmic tail. (B) Selection of the B cell epitope peptide sequence using the IEDB based on the epitope prediction, surface accessibility and antigenicity index. (C) ELISA analysis showing the generation of MERS-CoV Spike-492- or Spike-492 (L506F)-specific antibodies using sera. Sera were isolated from BALB/c mice immunized with a complex of B cell epitope peptide (Spike-492 or Spike-492 (L506F)) and Lipoplex (O).</p>
</caption>
<graphic xlink:href="bmb-52-397f1"></graphic>
</fig>
<fig id="f2-bmb-52-397" orientation="portrait" position="float"><label>Fig. 2</label>
<caption><p>Purification and characterization of anti-MERS-CoV Spike-492 (L506F) epitope-specific monoclonal antibody. (A) Purified 506-2G10G5 monoclonal antibody was analyzed by SDS-PAGE. R, reducing condition; NR, non-reducing conditions. (B) Isotype of the purified 506-2G10G5 monoclonal antibody was determined by ELISA. (C) The binding affinity of the 506-2G10G5 monoclonal antibody to the MERS-CoV Spike-492 (L506F) peptide was measured by ELISA, and the EC<sub>50</sub>
value was evaluated with the Sigma Plot program. (D–F) MERS-CoV-infected and non-infected Vero cells lysates were treated with PBS (−) or PNGase F (+). The lysates were immunoblotted with the 506-2G10G5 monoclonal antibody (D). Western blot analysis using the 506-2G10G5 monoclonal antibody was performed following the immunoprecipitation of lysates with the 506-2G10G5 monoclonal antibody (E). Lysates were subjected to western blotting with an anti-β actin antibody (F). (G, H) Cross-reactivity of the monoclonal antibodies. 96-well immunoplates coated with the MERS-CoV Spike-492 (G) or Spike-492 (L506F) (H) peptides and incubated with either the 492-1G10E4E2 or 506-2G10G5 monoclonal antibody. The reactivity of the antibodies to each peptide was determined by ELISA assay.</p>
</caption>
<graphic xlink:href="bmb-52-397f2"></graphic>
</fig>
<fig id="f3-bmb-52-397" orientation="portrait" position="float"><label>Fig. 3</label>
<caption><p>Immunofluorescence assay and confocal microscopy for the detection of MERS-CoV-infected cells with the Spike-492 (L506F) monoclonal antibody. (A) Indirect immunofluorescence assay. A mixture of MERS-CoV-infected and non-infected Vero cells was stained with the 506-2G10G5 monoclonal antibody or normal mouse IgG, followed by incubation with the Alexa488-conjugated goat anti-mouse IgG antibody (green). Cells were counterstained with Hoechst 33258 for nuclear staining (blue). Images were taken using a fluorescence microscope. (B) Confocal microscopy. Vero cells were infected with MERS-CoV for 2 days. The cells were stained with the 506-2G10G5 monoclonal antibody or normal mouse IgG and then incubated with the Alexa488-conjugated goat anti-mouse IgG antibody (green). Nuclei were stained using Hoechst 33258 (blue). Scale bar, 10 μm.</p>
</caption>
<graphic xlink:href="bmb-52-397f3"></graphic>
</fig>
<fig id="f4-bmb-52-397" orientation="portrait" position="float"><label>Fig. 4</label>
<caption><p>Inhibition of MERS-CoV infection by the 492-1G10E4E2 and 506-2G10G5 monoclonal antibody. MERS-CoV was pre-incubated with two fold serially diluted normal mouse IgG, 492-1G10E4E2 or 506-2G10G5 monoclonal antibody for 30 min at 37°C. The virus-antibody mixture was added to the Vero cells and incubated for 1 h. After the incubation, the medium was replaced with DMEM/F12 containing 0.6% oxoid agar. The plaques were stained with crystal violet 4 days after infection. (A) A representative picture showing the plaque reduction assay. (B) Quatification of the plaque reduction assay against MERS-CoV after treatment with 100 μg/well to 0 μg/well of normal mouse IgG or 492-1G10E4E2 or 506-2G10G5 monoclonal antibody.</p>
</caption>
<graphic xlink:href="bmb-52-397f4"></graphic>
</fig>
</floats-group>
</pmc>
<affiliations><list><country><li>Corée du Sud</li>
</country>
</list>
<tree><country name="Corée du Sud"><noRegion><name sortKey="Park, Byoung Kwon" sort="Park, Byoung Kwon" uniqKey="Park B" first="Byoung Kwon" last="Park">Byoung Kwon Park</name>
</noRegion>
<name sortKey="Bae, Joon Yong" sort="Bae, Joon Yong" uniqKey="Bae J" first="Joon-Yong" last="Bae">Joon-Yong Bae</name>
<name sortKey="Kim, Jinsoo" sort="Kim, Jinsoo" uniqKey="Kim J" first="Jinsoo" last="Kim">Jinsoo Kim</name>
<name sortKey="Kwon, Hyung Joo" sort="Kwon, Hyung Joo" uniqKey="Kwon H" first="Hyung-Joo" last="Kwon">Hyung-Joo Kwon</name>
<name sortKey="Kwon, Hyung Joo" sort="Kwon, Hyung Joo" uniqKey="Kwon H" first="Hyung-Joo" last="Kwon">Hyung-Joo Kwon</name>
<name sortKey="Lee, Su In" sort="Lee, Su In" uniqKey="Lee S" first="Su In" last="Lee">Su In Lee</name>
<name sortKey="Maharjan, Sony" sort="Maharjan, Sony" uniqKey="Maharjan S" first="Sony" last="Maharjan">Sony Maharjan</name>
<name sortKey="Park, Man Seong" sort="Park, Man Seong" uniqKey="Park M" first="Man-Seong" last="Park">Man-Seong Park</name>
</country>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Pmc/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000302 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Checkpoint/biblio.hfd -nk 000302 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= MersV1 |flux= Pmc |étape= Checkpoint |type= RBID |clé= PMC:6605520 |texte= Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex }}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Checkpoint/RBID.i -Sk "pubmed:30355437" \ | HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Checkpoint/biblio.hfd \ | NlmPubMed2Wicri -a MersV1
![]() | This area was generated with Dilib version V0.6.33. | ![]() |