Characterization of monoclonal antibodies against duck Tembusu virus E protein: an antigen-capture ELISA for the detection of Tembusu virus infection
The E protein of flaviviruses is the primary antigen that induces protective immunity, but a monoclonal antibody (mAb) against the E protein of duck Tembusu virus (DTMUV) has never been characterized. Six hybridoma cell lines secreting DTMUV anti-E mAbs were prepared and designated 2A5, 1F3, 1G2, 1B...
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| Veröffentlicht in: | Archives of virology 2015-03, Vol.160 (3), p.757-764 |
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| creator | Bai, Xiaofei Shaozhou, Wulin Zhang, Qingshan Li, Chenxi Qiu, Na Meng, Runzhe Liu, Ming Zhang, Yun |
| description | The E protein of flaviviruses is the primary antigen that induces protective immunity, but a monoclonal antibody (mAb) against the E protein of duck Tembusu virus (DTMUV) has never been characterized. Six hybridoma cell lines secreting DTMUV anti-E mAbs were prepared and designated 2A5, 1F3, 1G2, 1B11, 3B6, and 4F9, respectively. An immunofluorescence assay indicated that the mAbs could specifically bind to duck embryo fibroblast (DEF) cells infected with DTMUV and that the E protein was distributed in the cytoplasm of the infected cells. Immunoglobulin isotyping differentiated the mAbs as IgG1 (1G2, 1B11, 4F9, 1F3, and 2A5) and IgG2b (3B6). The mAbs were used to identify three epitopes, A (2A5, 1F3, and 1G2), B (1B11 and 4F9), and C (3B6) on the E protein on the basis of a competitive binding assay. By using mAbs 1F3 and 3B6, we developed an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) to detect E antigen from clinical samples. The AC-ELISA did not react with other known pathogens, indicating that the mAbs are specific for DTMUV. Compared to RT-PCR, the specificity and sensitivity of the AC-ELISA was 94.1 % and 98.0 %, respectively. This AC-ELISA thus represents a sensitive and rapid method for detecting DTMUV infection in birds. |
| doi_str_mv | 10.1007/s00705-014-2312-z |
| format | Article |
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Six hybridoma cell lines secreting DTMUV anti-E mAbs were prepared and designated 2A5, 1F3, 1G2, 1B11, 3B6, and 4F9, respectively. An immunofluorescence assay indicated that the mAbs could specifically bind to duck embryo fibroblast (DEF) cells infected with DTMUV and that the E protein was distributed in the cytoplasm of the infected cells. Immunoglobulin isotyping differentiated the mAbs as IgG1 (1G2, 1B11, 4F9, 1F3, and 2A5) and IgG2b (3B6). The mAbs were used to identify three epitopes, A (2A5, 1F3, and 1G2), B (1B11 and 4F9), and C (3B6) on the E protein on the basis of a competitive binding assay. By using mAbs 1F3 and 3B6, we developed an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) to detect E antigen from clinical samples. The AC-ELISA did not react with other known pathogens, indicating that the mAbs are specific for DTMUV. Compared to RT-PCR, the specificity and sensitivity of the AC-ELISA was 94.1 % and 98.0 %, respectively. This AC-ELISA thus represents a sensitive and rapid method for detecting DTMUV infection in birds.</description><identifier>ISSN: 0304-8608</identifier><identifier>EISSN: 1432-8798</identifier><identifier>DOI: 10.1007/s00705-014-2312-z</identifier><identifier>PMID: 25588821</identifier><language>eng</language><publisher>Vienna: Springer Vienna</publisher><subject>Animals ; Antibodies, Monoclonal - isolation & purification ; Antibodies, Viral - isolation & purification ; Antigens ; Antigens, Viral - analysis ; Biomedical and Life Sciences ; Biomedicine ; Bird Diseases - diagnosis ; Bird Diseases - virology ; Birds ; Cloning ; Ducks ; Enzyme-Linked Immunosorbent Assay - methods ; Enzymes ; Epitope Mapping ; Ethics ; Flavivirus - immunology ; Flavivirus - isolation & purification ; Flavivirus Infections - diagnosis ; Flavivirus Infections - veterinary ; Flavivirus Infections - virology ; Infections ; Infectious Diseases ; Laboratory animals ; Medical Microbiology ; Monoclonal antibodies ; Original Article ; Proteins ; Sensitivity and Specificity ; Tembusu virus ; Veterinary Medicine - methods ; Viral Envelope Proteins - analysis ; Virology ; Virology - methods ; Viruses</subject><ispartof>Archives of virology, 2015-03, Vol.160 (3), p.757-764</ispartof><rights>Springer-Verlag Wien 2015</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c475t-b27323bb15c64f37f87fb13c2dce453c79db1743375ea9523f17dc8b9cfc7723</citedby><cites>FETCH-LOGICAL-c475t-b27323bb15c64f37f87fb13c2dce453c79db1743375ea9523f17dc8b9cfc7723</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00705-014-2312-z$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00705-014-2312-z$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,777,781,27905,27906,41469,42538,51300</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25588821$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bai, Xiaofei</creatorcontrib><creatorcontrib>Shaozhou, Wulin</creatorcontrib><creatorcontrib>Zhang, Qingshan</creatorcontrib><creatorcontrib>Li, Chenxi</creatorcontrib><creatorcontrib>Qiu, Na</creatorcontrib><creatorcontrib>Meng, Runzhe</creatorcontrib><creatorcontrib>Liu, Ming</creatorcontrib><creatorcontrib>Zhang, Yun</creatorcontrib><title>Characterization of monoclonal antibodies against duck Tembusu virus E protein: an antigen-capture ELISA for the detection of Tembusu virus infection</title><title>Archives of virology</title><addtitle>Arch Virol</addtitle><addtitle>Arch Virol</addtitle><description>The E protein of flaviviruses is the primary antigen that induces protective immunity, but a monoclonal antibody (mAb) against the E protein of duck Tembusu virus (DTMUV) has never been characterized. Six hybridoma cell lines secreting DTMUV anti-E mAbs were prepared and designated 2A5, 1F3, 1G2, 1B11, 3B6, and 4F9, respectively. An immunofluorescence assay indicated that the mAbs could specifically bind to duck embryo fibroblast (DEF) cells infected with DTMUV and that the E protein was distributed in the cytoplasm of the infected cells. Immunoglobulin isotyping differentiated the mAbs as IgG1 (1G2, 1B11, 4F9, 1F3, and 2A5) and IgG2b (3B6). The mAbs were used to identify three epitopes, A (2A5, 1F3, and 1G2), B (1B11 and 4F9), and C (3B6) on the E protein on the basis of a competitive binding assay. By using mAbs 1F3 and 3B6, we developed an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) to detect E antigen from clinical samples. The AC-ELISA did not react with other known pathogens, indicating that the mAbs are specific for DTMUV. Compared to RT-PCR, the specificity and sensitivity of the AC-ELISA was 94.1 % and 98.0 %, respectively. This AC-ELISA thus represents a sensitive and rapid method for detecting DTMUV infection in birds.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - isolation & purification</subject><subject>Antibodies, Viral - isolation & purification</subject><subject>Antigens</subject><subject>Antigens, Viral - analysis</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Bird Diseases - diagnosis</subject><subject>Bird Diseases - virology</subject><subject>Birds</subject><subject>Cloning</subject><subject>Ducks</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Enzymes</subject><subject>Epitope Mapping</subject><subject>Ethics</subject><subject>Flavivirus - immunology</subject><subject>Flavivirus - isolation & purification</subject><subject>Flavivirus Infections - diagnosis</subject><subject>Flavivirus Infections - veterinary</subject><subject>Flavivirus Infections - virology</subject><subject>Infections</subject><subject>Infectious Diseases</subject><subject>Laboratory animals</subject><subject>Medical Microbiology</subject><subject>Monoclonal antibodies</subject><subject>Original Article</subject><subject>Proteins</subject><subject>Sensitivity and Specificity</subject><subject>Tembusu virus</subject><subject>Veterinary Medicine - methods</subject><subject>Viral Envelope Proteins - analysis</subject><subject>Virology</subject><subject>Virology - methods</subject><subject>Viruses</subject><issn>0304-8608</issn><issn>1432-8798</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNkc9uVCEYxYnR2LH6AG4MiRs3KH8uA-OumUxrk0lcOHsCXJhS74URuE2c9_B9ZXpboyYmbiDh-51zvnAAeE3we4Kx-FDagTnCpEOUEYqOT8CCdIwiKVbyKVhghjskl1iegRel3GLcHhh_Ds4o51JKShbgx_pGZ22ry-Goa0gRJg_HFJMdUtQD1LEGk_rgCtR7HWKpsJ_sV7hzo5nKBO9CngrcwENO1YX4sQnuNXsXkdWHOmUHN9vrLxfQpwzrjYO9q84-Jv1pE6KfRy_BM6-H4l493Odgd7nZrT-h7eer6_XFFtlO8IoMFYwyYwi3y84z4aXwhjBLe-s6zqxY9YaIjjHBnV5xyjwRvZVmZb0VgrJz8G62bdt_m1ypagzFumHQ0aWpKLJcStpxyfh_oFx2pyzS0Ld_obdpyu0z7ynRTLkUjSIzZXMqJTuvDjmMOn9XBKtTu2puV7V21alddWyaNw_Okxld_0vxWGcD6AyUNop7l3-L_qfrT_2fsa4</recordid><startdate>20150301</startdate><enddate>20150301</enddate><creator>Bai, Xiaofei</creator><creator>Shaozhou, Wulin</creator><creator>Zhang, Qingshan</creator><creator>Li, Chenxi</creator><creator>Qiu, Na</creator><creator>Meng, Runzhe</creator><creator>Liu, Ming</creator><creator>Zhang, Yun</creator><general>Springer Vienna</general><general>Springer Nature B.V</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>3V.</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</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>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20150301</creationdate><title>Characterization of monoclonal antibodies against duck Tembusu virus E protein: an antigen-capture ELISA for the detection of Tembusu virus infection</title><author>Bai, Xiaofei ; Shaozhou, Wulin ; Zhang, Qingshan ; Li, Chenxi ; Qiu, Na ; Meng, Runzhe ; Liu, Ming ; Zhang, Yun</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c475t-b27323bb15c64f37f87fb13c2dce453c79db1743375ea9523f17dc8b9cfc7723</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - isolation & purification</topic><topic>Antibodies, Viral - isolation & purification</topic><topic>Antigens</topic><topic>Antigens, Viral - analysis</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Bird Diseases - diagnosis</topic><topic>Bird Diseases - virology</topic><topic>Birds</topic><topic>Cloning</topic><topic>Ducks</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Enzymes</topic><topic>Epitope Mapping</topic><topic>Ethics</topic><topic>Flavivirus - immunology</topic><topic>Flavivirus - isolation & purification</topic><topic>Flavivirus Infections - diagnosis</topic><topic>Flavivirus Infections - veterinary</topic><topic>Flavivirus Infections - virology</topic><topic>Infections</topic><topic>Infectious Diseases</topic><topic>Laboratory animals</topic><topic>Medical Microbiology</topic><topic>Monoclonal antibodies</topic><topic>Original Article</topic><topic>Proteins</topic><topic>Sensitivity and Specificity</topic><topic>Tembusu virus</topic><topic>Veterinary Medicine - 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Academic</collection><jtitle>Archives of virology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bai, Xiaofei</au><au>Shaozhou, Wulin</au><au>Zhang, Qingshan</au><au>Li, Chenxi</au><au>Qiu, Na</au><au>Meng, Runzhe</au><au>Liu, Ming</au><au>Zhang, Yun</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization of monoclonal antibodies against duck Tembusu virus E protein: an antigen-capture ELISA for the detection of Tembusu virus infection</atitle><jtitle>Archives of virology</jtitle><stitle>Arch Virol</stitle><addtitle>Arch Virol</addtitle><date>2015-03-01</date><risdate>2015</risdate><volume>160</volume><issue>3</issue><spage>757</spage><epage>764</epage><pages>757-764</pages><issn>0304-8608</issn><eissn>1432-8798</eissn><abstract>The E protein of flaviviruses is the primary antigen that induces protective immunity, but a monoclonal antibody (mAb) against the E protein of duck Tembusu virus (DTMUV) has never been characterized. Six hybridoma cell lines secreting DTMUV anti-E mAbs were prepared and designated 2A5, 1F3, 1G2, 1B11, 3B6, and 4F9, respectively. An immunofluorescence assay indicated that the mAbs could specifically bind to duck embryo fibroblast (DEF) cells infected with DTMUV and that the E protein was distributed in the cytoplasm of the infected cells. Immunoglobulin isotyping differentiated the mAbs as IgG1 (1G2, 1B11, 4F9, 1F3, and 2A5) and IgG2b (3B6). The mAbs were used to identify three epitopes, A (2A5, 1F3, and 1G2), B (1B11 and 4F9), and C (3B6) on the E protein on the basis of a competitive binding assay. By using mAbs 1F3 and 3B6, we developed an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) to detect E antigen from clinical samples. The AC-ELISA did not react with other known pathogens, indicating that the mAbs are specific for DTMUV. Compared to RT-PCR, the specificity and sensitivity of the AC-ELISA was 94.1 % and 98.0 %, respectively. This AC-ELISA thus represents a sensitive and rapid method for detecting DTMUV infection in birds.</abstract><cop>Vienna</cop><pub>Springer Vienna</pub><pmid>25588821</pmid><doi>10.1007/s00705-014-2312-z</doi><tpages>8</tpages></addata></record> |
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| subjects | Animals Antibodies, Monoclonal - isolation & purification Antibodies, Viral - isolation & purification Antigens Antigens, Viral - analysis Biomedical and Life Sciences Biomedicine Bird Diseases - diagnosis Bird Diseases - virology Birds Cloning Ducks Enzyme-Linked Immunosorbent Assay - methods Enzymes Epitope Mapping Ethics Flavivirus - immunology Flavivirus - isolation & purification Flavivirus Infections - diagnosis Flavivirus Infections - veterinary Flavivirus Infections - virology Infections Infectious Diseases Laboratory animals Medical Microbiology Monoclonal antibodies Original Article Proteins Sensitivity and Specificity Tembusu virus Veterinary Medicine - methods Viral Envelope Proteins - analysis Virology Virology - methods Viruses |
| title | Characterization of monoclonal antibodies against duck Tembusu virus E protein: an antigen-capture ELISA for the detection of Tembusu virus infection |
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