Critical epitopes in the nucleocapsid protein of SFTS virus recognized by a panel of SFTS patients derived human monoclonal antibodies
SFTS virus (SFTSV) is a newly discovered pathogen to cause severe fever with thrombocytopenia syndrome (SFTS) in human. Successful control of SFTSV epidemic requires better understanding of the antigen target in humoral immune responses to the new bunyavirus infection. We have generated a combinator...
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| Veröffentlicht in: | PloS one 2012-06, Vol.7 (6), p.e38291 |
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| creator | Yu, Li Zhang, Li Sun, Lina Lu, Jing Wu, Wei Li, Chuan Zhang, Quanfu Zhang, Fushun Jin, Cong Wang, Xianjun Bi, Zhenqiang Li, Dexin Liang, Mifang |
| description | SFTS virus (SFTSV) is a newly discovered pathogen to cause severe fever with thrombocytopenia syndrome (SFTS) in human. Successful control of SFTSV epidemic requires better understanding of the antigen target in humoral immune responses to the new bunyavirus infection.
We have generated a combinatorial Fab antibody phage library from two SFTS patients recovered from SFTSV infection. To date, 94 unique human antibodies have been generated and characterized from over 1200 Fab antibody clones obtained by screening the library with SFTS purified virions. All those monoclonal antibodies (MAbs) recognized the nucleocapsid (N) protein of SFTSV while none of them were reactive to the viral glycoproteins Gn or Gc. Furthermore, over screening 1000 mouse monoclonal antibody clones derived from SFTSV virions immunization, 462 clones reacted with N protein, while only 16 clones were reactive to glycoprotein. Furthermore, epitope mapping of SFTSV N protein was performed through molecular simulation, site mutation and competitive ELISA, and we found that at least 4 distinct antigenic epitopes within N protein were recognized by those human and mouse MAbs, in particular mutation of Glu10 to Ala10 abolished or significantly reduced the binding activity of nearly most SFTS patients derived MAbs.
The large number of human recombinant MAbs derived from SFTS patients recognized the viral N protein indicated the important role of the N protein in humoral responses to SFTSV infection, and the critical epitopes we defined in this study provided molecular basis for detection and diagnosis of SFTSV infection. |
| doi_str_mv | 10.1371/journal.pone.0038291 |
| format | Article |
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We have generated a combinatorial Fab antibody phage library from two SFTS patients recovered from SFTSV infection. To date, 94 unique human antibodies have been generated and characterized from over 1200 Fab antibody clones obtained by screening the library with SFTS purified virions. All those monoclonal antibodies (MAbs) recognized the nucleocapsid (N) protein of SFTSV while none of them were reactive to the viral glycoproteins Gn or Gc. Furthermore, over screening 1000 mouse monoclonal antibody clones derived from SFTSV virions immunization, 462 clones reacted with N protein, while only 16 clones were reactive to glycoprotein. Furthermore, epitope mapping of SFTSV N protein was performed through molecular simulation, site mutation and competitive ELISA, and we found that at least 4 distinct antigenic epitopes within N protein were recognized by those human and mouse MAbs, in particular mutation of Glu10 to Ala10 abolished or significantly reduced the binding activity of nearly most SFTS patients derived MAbs.
The large number of human recombinant MAbs derived from SFTS patients recognized the viral N protein indicated the important role of the N protein in humoral responses to SFTSV infection, and the critical epitopes we defined in this study provided molecular basis for detection and diagnosis of SFTSV infection.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0038291</identifier><identifier>PMID: 22719874</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Animals ; Antibodies, Monoclonal - immunology ; Antigenic determinants ; Antigens ; Biology ; Blotting, Western ; Cloning ; Combinatorial analysis ; Disease control ; Disease prevention ; Enzyme-Linked Immunosorbent Assay ; Enzymes ; Epidemics ; Epitope Mapping ; Epitopes - genetics ; Epitopes - immunology ; Fab ; Fever ; Fluorescent Antibody Technique ; Genes ; Glycoproteins ; Health aspects ; Humans ; Immune response (humoral) ; Immunization ; Immunoassay ; Immunoglobulins ; Infection ; Infections ; Infectious diseases ; Libraries ; Medical laboratories ; Medicine ; Mice ; Models, Molecular ; Monoclonal antibodies ; Mutagenesis ; Mutagenesis, Site-Directed ; Mutation ; N protein ; Neutralization Tests ; Nucleocapsid Proteins - immunology ; Nucleocapsids ; Pathogenesis ; Patients ; Peptide mapping ; Phages ; Phlebovirus - immunology ; Proteins ; Screening ; Simulation ; Studies ; Thrombocytopenia ; Virions ; Viruses</subject><ispartof>PloS one, 2012-06, Vol.7 (6), p.e38291</ispartof><rights>COPYRIGHT 2012 Public Library of Science</rights><rights>2012 Yu et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>Yu et al. 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-56bbed6089a587c7998b89d318c606cd2e6f7e0e8905c2b98cc5c71e22931e733</citedby><cites>FETCH-LOGICAL-c692t-56bbed6089a587c7998b89d318c606cd2e6f7e0e8905c2b98cc5c71e22931e733</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3373585/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3373585/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79342,79343</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22719874$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yu, Li</creatorcontrib><creatorcontrib>Zhang, Li</creatorcontrib><creatorcontrib>Sun, Lina</creatorcontrib><creatorcontrib>Lu, Jing</creatorcontrib><creatorcontrib>Wu, Wei</creatorcontrib><creatorcontrib>Li, Chuan</creatorcontrib><creatorcontrib>Zhang, Quanfu</creatorcontrib><creatorcontrib>Zhang, Fushun</creatorcontrib><creatorcontrib>Jin, Cong</creatorcontrib><creatorcontrib>Wang, Xianjun</creatorcontrib><creatorcontrib>Bi, Zhenqiang</creatorcontrib><creatorcontrib>Li, Dexin</creatorcontrib><creatorcontrib>Liang, Mifang</creatorcontrib><title>Critical epitopes in the nucleocapsid protein of SFTS virus recognized by a panel of SFTS patients derived human monoclonal antibodies</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>SFTS virus (SFTSV) is a newly discovered pathogen to cause severe fever with thrombocytopenia syndrome (SFTS) in human. Successful control of SFTSV epidemic requires better understanding of the antigen target in humoral immune responses to the new bunyavirus infection.
We have generated a combinatorial Fab antibody phage library from two SFTS patients recovered from SFTSV infection. To date, 94 unique human antibodies have been generated and characterized from over 1200 Fab antibody clones obtained by screening the library with SFTS purified virions. All those monoclonal antibodies (MAbs) recognized the nucleocapsid (N) protein of SFTSV while none of them were reactive to the viral glycoproteins Gn or Gc. Furthermore, over screening 1000 mouse monoclonal antibody clones derived from SFTSV virions immunization, 462 clones reacted with N protein, while only 16 clones were reactive to glycoprotein. Furthermore, epitope mapping of SFTSV N protein was performed through molecular simulation, site mutation and competitive ELISA, and we found that at least 4 distinct antigenic epitopes within N protein were recognized by those human and mouse MAbs, in particular mutation of Glu10 to Ala10 abolished or significantly reduced the binding activity of nearly most SFTS patients derived MAbs.
The large number of human recombinant MAbs derived from SFTS patients recognized the viral N protein indicated the important role of the N protein in humoral responses to SFTSV infection, and the critical epitopes we defined in this study provided molecular basis for detection and diagnosis of SFTSV infection.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Antigenic determinants</subject><subject>Antigens</subject><subject>Biology</subject><subject>Blotting, Western</subject><subject>Cloning</subject><subject>Combinatorial analysis</subject><subject>Disease control</subject><subject>Disease prevention</subject><subject>Enzyme-Linked Immunosorbent Assay</subject><subject>Enzymes</subject><subject>Epidemics</subject><subject>Epitope Mapping</subject><subject>Epitopes - genetics</subject><subject>Epitopes - immunology</subject><subject>Fab</subject><subject>Fever</subject><subject>Fluorescent Antibody Technique</subject><subject>Genes</subject><subject>Glycoproteins</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Immune response (humoral)</subject><subject>Immunization</subject><subject>Immunoassay</subject><subject>Immunoglobulins</subject><subject>Infection</subject><subject>Infections</subject><subject>Infectious diseases</subject><subject>Libraries</subject><subject>Medical laboratories</subject><subject>Medicine</subject><subject>Mice</subject><subject>Models, Molecular</subject><subject>Monoclonal antibodies</subject><subject>Mutagenesis</subject><subject>Mutagenesis, Site-Directed</subject><subject>Mutation</subject><subject>N protein</subject><subject>Neutralization Tests</subject><subject>Nucleocapsid Proteins - immunology</subject><subject>Nucleocapsids</subject><subject>Pathogenesis</subject><subject>Patients</subject><subject>Peptide mapping</subject><subject>Phages</subject><subject>Phlebovirus - immunology</subject><subject>Proteins</subject><subject>Screening</subject><subject>Simulation</subject><subject>Studies</subject><subject>Thrombocytopenia</subject><subject>Virions</subject><subject>Viruses</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk99qFDEUxgdRbK2-gWhAELzYNX82k8xNoRSrC4WCrd6GTHJ2N8tsMiaZxfoAPrdpd1p2QUECScj5nS-Hj3Oq6jXBU8IE-bgOQ_S6m_bBwxRjJmlDnlTHpGF0UlPMnu7dj6oXKa0x5kzW9fPqiFJBGilmx9Xv8-iyM7pD0LscekjIeZRXgPxgOghG98lZ1MeQoQTCAl1f3FyjrYtDQhFMWHr3Cyxqb5FGvfbQPTK9zg58TshCdNvCrIaN9mgTfDBdKKUj7bNrg3WQXlbPFrpL8Go8T6pvF59uzr9MLq8-z8_PLiembmie8LptwdZYNppLYUTTyFY2lhFpalwbS6FeCMAgG8wNbRtpDDeCAKUNIyAYO6ne7nT7LiQ1WpgUYZRjKjCVhZjvCBv0WvXRbXS8VUE7df8Q4lLpWBzrQGHaWA21JpTIGWdccyusbcvGWyrM3W-n429DuwFrihtRdweihxHvVmoZtooxwbjkReDdKBDDjwFS_kfJI7XUpSrnF6GImY1LRp3NhCCEs3tq-heqLAsbZ0oTLVx5P0j4cJBQmAw_81IPKan59df_Z6--H7Lv99gV6C6vUuiG7IJPh-BsB5oYUoqweHSOYHU3Aw9uqLsZUOMMlLQ3-64_Jj00PfsDTbcC8A</recordid><startdate>20120612</startdate><enddate>20120612</enddate><creator>Yu, Li</creator><creator>Zhang, Li</creator><creator>Sun, Lina</creator><creator>Lu, Jing</creator><creator>Wu, Wei</creator><creator>Li, Chuan</creator><creator>Zhang, Quanfu</creator><creator>Zhang, Fushun</creator><creator>Jin, Cong</creator><creator>Wang, Xianjun</creator><creator>Bi, Zhenqiang</creator><creator>Li, Dexin</creator><creator>Liang, Mifang</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20120612</creationdate><title>Critical epitopes in the nucleocapsid protein of SFTS virus recognized by a panel of SFTS patients derived human monoclonal antibodies</title><author>Yu, Li ; Zhang, Li ; Sun, Lina ; Lu, Jing ; Wu, Wei ; Li, Chuan ; Zhang, Quanfu ; Zhang, Fushun ; Jin, Cong ; Wang, Xianjun ; Bi, Zhenqiang ; Li, Dexin ; Liang, Mifang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-56bbed6089a587c7998b89d318c606cd2e6f7e0e8905c2b98cc5c71e22931e733</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - 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Successful control of SFTSV epidemic requires better understanding of the antigen target in humoral immune responses to the new bunyavirus infection.
We have generated a combinatorial Fab antibody phage library from two SFTS patients recovered from SFTSV infection. To date, 94 unique human antibodies have been generated and characterized from over 1200 Fab antibody clones obtained by screening the library with SFTS purified virions. All those monoclonal antibodies (MAbs) recognized the nucleocapsid (N) protein of SFTSV while none of them were reactive to the viral glycoproteins Gn or Gc. Furthermore, over screening 1000 mouse monoclonal antibody clones derived from SFTSV virions immunization, 462 clones reacted with N protein, while only 16 clones were reactive to glycoprotein. Furthermore, epitope mapping of SFTSV N protein was performed through molecular simulation, site mutation and competitive ELISA, and we found that at least 4 distinct antigenic epitopes within N protein were recognized by those human and mouse MAbs, in particular mutation of Glu10 to Ala10 abolished or significantly reduced the binding activity of nearly most SFTS patients derived MAbs.
The large number of human recombinant MAbs derived from SFTS patients recognized the viral N protein indicated the important role of the N protein in humoral responses to SFTSV infection, and the critical epitopes we defined in this study provided molecular basis for detection and diagnosis of SFTSV infection.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>22719874</pmid><doi>10.1371/journal.pone.0038291</doi><tpages>e38291</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2012-06, Vol.7 (6), p.e38291 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1325027028 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Animals Antibodies, Monoclonal - immunology Antigenic determinants Antigens Biology Blotting, Western Cloning Combinatorial analysis Disease control Disease prevention Enzyme-Linked Immunosorbent Assay Enzymes Epidemics Epitope Mapping Epitopes - genetics Epitopes - immunology Fab Fever Fluorescent Antibody Technique Genes Glycoproteins Health aspects Humans Immune response (humoral) Immunization Immunoassay Immunoglobulins Infection Infections Infectious diseases Libraries Medical laboratories Medicine Mice Models, Molecular Monoclonal antibodies Mutagenesis Mutagenesis, Site-Directed Mutation N protein Neutralization Tests Nucleocapsid Proteins - immunology Nucleocapsids Pathogenesis Patients Peptide mapping Phages Phlebovirus - immunology Proteins Screening Simulation Studies Thrombocytopenia Virions Viruses |
| title | Critical epitopes in the nucleocapsid protein of SFTS virus recognized by a panel of SFTS patients derived human monoclonal antibodies |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-05T20%3A07%3A56IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Critical%20epitopes%20in%20the%20nucleocapsid%20protein%20of%20SFTS%20virus%20recognized%20by%20a%20panel%20of%20SFTS%20patients%20derived%20human%20monoclonal%20antibodies&rft.jtitle=PloS%20one&rft.au=Yu,%20Li&rft.date=2012-06-12&rft.volume=7&rft.issue=6&rft.spage=e38291&rft.pages=e38291-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0038291&rft_dat=%3Cgale_plos_%3EA477115328%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1325027028&rft_id=info:pmid/22719874&rft_galeid=A477115328&rft_doaj_id=oai_doaj_org_article_029dae6a12184535a5d7ddbd7d5b27c3&rfr_iscdi=true |