Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis
► Properly folded NS1 protein expressed in E. coli is suitable for DENV diagnosis. ► Oligomerization of NS1 is important to its immunogenicity. ► Anti-NS1 polyclonal antibodies are able to detect NS1 in DENV-positive human serum. The non-structural 1 (NS1) protein plays an important role in dengue d...
Gespeichert in:
| Veröffentlicht in: | Journal of virological methods 2011-07, Vol.175 (1), p.109-116 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 116 |
|---|---|
| container_issue | 1 |
| container_start_page | 109 |
| container_title | Journal of virological methods |
| container_volume | 175 |
| creator | Allonso, Diego da Silva Rosa, Marcela Coelho, Diego Rodrigues da Costa, Simone Morais Nogueira, Rita Maria Ribeiro Bozza, Fernando Augusto Santos, Flavia Barreto Dos de Barcelos Alves, Ada Maria Mohana-Borges, Ronaldo |
| description | ► Properly folded NS1 protein expressed in E. coli is suitable for DENV diagnosis. ► Oligomerization of NS1 is important to its immunogenicity. ► Anti-NS1 polyclonal antibodies are able to detect NS1 in DENV-positive human serum.
The non-structural 1 (NS1) protein plays an important role in dengue diagnosis because it has been detected as a soluble serum antigen in both primary and secondary infections. The NS1 protein was expressed in Escherichia coli cells, and the efficiency of four different refolding protocols was tested. All of the protocols generated dimeric NS1 in a conformation similar to that of the protein expressed by eukaryotic cells. A polyclonal antibody produced from the properly folded E. coli recombinant NS1 (rNS1) protein proved to be a useful tool for the diagnosis of Dengue virus because it detected 100% of the Dengue virus 2 (DENV2) in infected patients’ sera and 60% of the DENV IgM-positive sera not detected by commercial NS1-based diagnostic kits. These data suggest a high-efficiency method for correctly folding rNS1 that maintains its structural and immunogenic properties. In addition, a detection method using the polyclonal antibody against correctly folded rNS1 seemed to be more sensitive and efficient for NS1 detection in serum, highlighting its usefulness for developing a high-sensitivity diagnostic kit. |
| doi_str_mv | 10.1016/j.jviromet.2011.04.029 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_907166544</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0166093411001674</els_id><sourcerecordid>871004255</sourcerecordid><originalsourceid>FETCH-LOGICAL-c477t-aefc1ecff1e1208fed24d119ed3643f00bc9145680c2165f2abe499e30e5796a3</originalsourceid><addsrcrecordid>eNqFkcFuEzEQhi0EoqHwCpUviFOW8a7Xu76B2kIrVYAEnC3HHkeOHDvYuxF5Al4bR0nh2NN4pG_-fzw_IVcMGgZMvN80m73PaYtT0wJjDfAGWvmMLNg4yCXIkT8niwqK-u74BXlVygYA-qHrXpKLlvVCDlIsyJ9vKRxMSFEHquPkV8l6LFSvtY9lorucdpjDgboULFp6g3E9I63Wc6FfvrMjMKGPFH_vMpZSkdrcNtSk4ClGvQpIC8biJ7_H6mAp6qOePQlZr9cxFV9ekxdOh4JvzvWS_Px0--P6bvnw9fP99ceHpeHDMC01OsPQOMeQtTA6tC23jEm0neCdA1gZyXgvRjAtE71r9Qq5lNgB9vW_ursk7066dfFfM5ZJbX0xGIKOmOaiJAz1aD3nT5LjwAB42_eVFCfS5FRKRqd22W91PigG6piW2qjHtNQxLQVc1bTq4NXZYl5t0f4be4ynAm_PgC5GB5d1NL7853grRyGgch9OHNbT7T1mVYzHaND6jGZSNvmndvkLE8m5FQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>871004255</pqid></control><display><type>article</type><title>Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals Complete</source><creator>Allonso, Diego ; da Silva Rosa, Marcela ; Coelho, Diego Rodrigues ; da Costa, Simone Morais ; Nogueira, Rita Maria Ribeiro ; Bozza, Fernando Augusto ; Santos, Flavia Barreto Dos ; de Barcelos Alves, Ada Maria ; Mohana-Borges, Ronaldo</creator><creatorcontrib>Allonso, Diego ; da Silva Rosa, Marcela ; Coelho, Diego Rodrigues ; da Costa, Simone Morais ; Nogueira, Rita Maria Ribeiro ; Bozza, Fernando Augusto ; Santos, Flavia Barreto Dos ; de Barcelos Alves, Ada Maria ; Mohana-Borges, Ronaldo</creatorcontrib><description>► Properly folded NS1 protein expressed in E. coli is suitable for DENV diagnosis. ► Oligomerization of NS1 is important to its immunogenicity. ► Anti-NS1 polyclonal antibodies are able to detect NS1 in DENV-positive human serum.
The non-structural 1 (NS1) protein plays an important role in dengue diagnosis because it has been detected as a soluble serum antigen in both primary and secondary infections. The NS1 protein was expressed in Escherichia coli cells, and the efficiency of four different refolding protocols was tested. All of the protocols generated dimeric NS1 in a conformation similar to that of the protein expressed by eukaryotic cells. A polyclonal antibody produced from the properly folded E. coli recombinant NS1 (rNS1) protein proved to be a useful tool for the diagnosis of Dengue virus because it detected 100% of the Dengue virus 2 (DENV2) in infected patients’ sera and 60% of the DENV IgM-positive sera not detected by commercial NS1-based diagnostic kits. These data suggest a high-efficiency method for correctly folding rNS1 that maintains its structural and immunogenic properties. In addition, a detection method using the polyclonal antibody against correctly folded rNS1 seemed to be more sensitive and efficient for NS1 detection in serum, highlighting its usefulness for developing a high-sensitivity diagnostic kit.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/j.jviromet.2011.04.029</identifier><identifier>PMID: 21569796</identifier><identifier>CODEN: JVMEDH</identifier><language>eng</language><publisher>Kidlington: Elsevier B.V</publisher><subject>Animals ; Antibodies, Viral - blood ; Antibodies, Viral - immunology ; Biological and medical sciences ; Blotting, Western ; Dengue - diagnosis ; Dengue virus ; Dengue Virus - genetics ; Dengue Virus - immunology ; Dengue virus type 2 ; E. coli ; Early Diagnosis ; Enzyme-Linked Immunosorbent Assay - methods ; Escherichia coli ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Fundamental and applied biological sciences. Psychology ; Glycoproteins - immunology ; Humans ; Mice ; Mice, Inbred BALB C ; Microbiology ; NS1 protein ; Protein Folding ; Rabbits ; Techniques used in virology ; Viral Nonstructural Proteins - genetics ; Viral Nonstructural Proteins - immunology ; Viral Nonstructural Proteins - metabolism ; Virology</subject><ispartof>Journal of virological methods, 2011-07, Vol.175 (1), p.109-116</ispartof><rights>2011 Elsevier B.V.</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2011 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c477t-aefc1ecff1e1208fed24d119ed3643f00bc9145680c2165f2abe499e30e5796a3</citedby><cites>FETCH-LOGICAL-c477t-aefc1ecff1e1208fed24d119ed3643f00bc9145680c2165f2abe499e30e5796a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jviromet.2011.04.029$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24298660$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21569796$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Allonso, Diego</creatorcontrib><creatorcontrib>da Silva Rosa, Marcela</creatorcontrib><creatorcontrib>Coelho, Diego Rodrigues</creatorcontrib><creatorcontrib>da Costa, Simone Morais</creatorcontrib><creatorcontrib>Nogueira, Rita Maria Ribeiro</creatorcontrib><creatorcontrib>Bozza, Fernando Augusto</creatorcontrib><creatorcontrib>Santos, Flavia Barreto Dos</creatorcontrib><creatorcontrib>de Barcelos Alves, Ada Maria</creatorcontrib><creatorcontrib>Mohana-Borges, Ronaldo</creatorcontrib><title>Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis</title><title>Journal of virological methods</title><addtitle>J Virol Methods</addtitle><description>► Properly folded NS1 protein expressed in E. coli is suitable for DENV diagnosis. ► Oligomerization of NS1 is important to its immunogenicity. ► Anti-NS1 polyclonal antibodies are able to detect NS1 in DENV-positive human serum.
The non-structural 1 (NS1) protein plays an important role in dengue diagnosis because it has been detected as a soluble serum antigen in both primary and secondary infections. The NS1 protein was expressed in Escherichia coli cells, and the efficiency of four different refolding protocols was tested. All of the protocols generated dimeric NS1 in a conformation similar to that of the protein expressed by eukaryotic cells. A polyclonal antibody produced from the properly folded E. coli recombinant NS1 (rNS1) protein proved to be a useful tool for the diagnosis of Dengue virus because it detected 100% of the Dengue virus 2 (DENV2) in infected patients’ sera and 60% of the DENV IgM-positive sera not detected by commercial NS1-based diagnostic kits. These data suggest a high-efficiency method for correctly folding rNS1 that maintains its structural and immunogenic properties. In addition, a detection method using the polyclonal antibody against correctly folded rNS1 seemed to be more sensitive and efficient for NS1 detection in serum, highlighting its usefulness for developing a high-sensitivity diagnostic kit.</description><subject>Animals</subject><subject>Antibodies, Viral - blood</subject><subject>Antibodies, Viral - immunology</subject><subject>Biological and medical sciences</subject><subject>Blotting, Western</subject><subject>Dengue - diagnosis</subject><subject>Dengue virus</subject><subject>Dengue Virus - genetics</subject><subject>Dengue Virus - immunology</subject><subject>Dengue virus type 2</subject><subject>E. coli</subject><subject>Early Diagnosis</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glycoproteins - immunology</subject><subject>Humans</subject><subject>Mice</subject><subject>Mice, Inbred BALB C</subject><subject>Microbiology</subject><subject>NS1 protein</subject><subject>Protein Folding</subject><subject>Rabbits</subject><subject>Techniques used in virology</subject><subject>Viral Nonstructural Proteins - genetics</subject><subject>Viral Nonstructural Proteins - immunology</subject><subject>Viral Nonstructural Proteins - metabolism</subject><subject>Virology</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcFuEzEQhi0EoqHwCpUviFOW8a7Xu76B2kIrVYAEnC3HHkeOHDvYuxF5Al4bR0nh2NN4pG_-fzw_IVcMGgZMvN80m73PaYtT0wJjDfAGWvmMLNg4yCXIkT8niwqK-u74BXlVygYA-qHrXpKLlvVCDlIsyJ9vKRxMSFEHquPkV8l6LFSvtY9lorucdpjDgboULFp6g3E9I63Wc6FfvrMjMKGPFH_vMpZSkdrcNtSk4ClGvQpIC8biJ7_H6mAp6qOePQlZr9cxFV9ekxdOh4JvzvWS_Px0--P6bvnw9fP99ceHpeHDMC01OsPQOMeQtTA6tC23jEm0neCdA1gZyXgvRjAtE71r9Qq5lNgB9vW_ursk7066dfFfM5ZJbX0xGIKOmOaiJAz1aD3nT5LjwAB42_eVFCfS5FRKRqd22W91PigG6piW2qjHtNQxLQVc1bTq4NXZYl5t0f4be4ynAm_PgC5GB5d1NL7853grRyGgch9OHNbT7T1mVYzHaND6jGZSNvmndvkLE8m5FQ</recordid><startdate>20110701</startdate><enddate>20110701</enddate><creator>Allonso, Diego</creator><creator>da Silva Rosa, Marcela</creator><creator>Coelho, Diego Rodrigues</creator><creator>da Costa, Simone Morais</creator><creator>Nogueira, Rita Maria Ribeiro</creator><creator>Bozza, Fernando Augusto</creator><creator>Santos, Flavia Barreto Dos</creator><creator>de Barcelos Alves, Ada Maria</creator><creator>Mohana-Borges, Ronaldo</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>6I.</scope><scope>AAFTH</scope><scope>IQODW</scope><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>7X8</scope><scope>7U9</scope><scope>C1K</scope><scope>F1W</scope><scope>H94</scope><scope>H95</scope><scope>H97</scope><scope>L.G</scope></search><sort><creationdate>20110701</creationdate><title>Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis</title><author>Allonso, Diego ; da Silva Rosa, Marcela ; Coelho, Diego Rodrigues ; da Costa, Simone Morais ; Nogueira, Rita Maria Ribeiro ; Bozza, Fernando Augusto ; Santos, Flavia Barreto Dos ; de Barcelos Alves, Ada Maria ; Mohana-Borges, Ronaldo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c477t-aefc1ecff1e1208fed24d119ed3643f00bc9145680c2165f2abe499e30e5796a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animals</topic><topic>Antibodies, Viral - blood</topic><topic>Antibodies, Viral - immunology</topic><topic>Biological and medical sciences</topic><topic>Blotting, Western</topic><topic>Dengue - diagnosis</topic><topic>Dengue virus</topic><topic>Dengue Virus - genetics</topic><topic>Dengue Virus - immunology</topic><topic>Dengue virus type 2</topic><topic>E. coli</topic><topic>Early Diagnosis</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glycoproteins - immunology</topic><topic>Humans</topic><topic>Mice</topic><topic>Mice, Inbred BALB C</topic><topic>Microbiology</topic><topic>NS1 protein</topic><topic>Protein Folding</topic><topic>Rabbits</topic><topic>Techniques used in virology</topic><topic>Viral Nonstructural Proteins - genetics</topic><topic>Viral Nonstructural Proteins - immunology</topic><topic>Viral Nonstructural Proteins - metabolism</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Allonso, Diego</creatorcontrib><creatorcontrib>da Silva Rosa, Marcela</creatorcontrib><creatorcontrib>Coelho, Diego Rodrigues</creatorcontrib><creatorcontrib>da Costa, Simone Morais</creatorcontrib><creatorcontrib>Nogueira, Rita Maria Ribeiro</creatorcontrib><creatorcontrib>Bozza, Fernando Augusto</creatorcontrib><creatorcontrib>Santos, Flavia Barreto Dos</creatorcontrib><creatorcontrib>de Barcelos Alves, Ada Maria</creatorcontrib><creatorcontrib>Mohana-Borges, Ronaldo</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Virology and AIDS Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Allonso, Diego</au><au>da Silva Rosa, Marcela</au><au>Coelho, Diego Rodrigues</au><au>da Costa, Simone Morais</au><au>Nogueira, Rita Maria Ribeiro</au><au>Bozza, Fernando Augusto</au><au>Santos, Flavia Barreto Dos</au><au>de Barcelos Alves, Ada Maria</au><au>Mohana-Borges, Ronaldo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>2011-07-01</date><risdate>2011</risdate><volume>175</volume><issue>1</issue><spage>109</spage><epage>116</epage><pages>109-116</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><coden>JVMEDH</coden><abstract>► Properly folded NS1 protein expressed in E. coli is suitable for DENV diagnosis. ► Oligomerization of NS1 is important to its immunogenicity. ► Anti-NS1 polyclonal antibodies are able to detect NS1 in DENV-positive human serum.
The non-structural 1 (NS1) protein plays an important role in dengue diagnosis because it has been detected as a soluble serum antigen in both primary and secondary infections. The NS1 protein was expressed in Escherichia coli cells, and the efficiency of four different refolding protocols was tested. All of the protocols generated dimeric NS1 in a conformation similar to that of the protein expressed by eukaryotic cells. A polyclonal antibody produced from the properly folded E. coli recombinant NS1 (rNS1) protein proved to be a useful tool for the diagnosis of Dengue virus because it detected 100% of the Dengue virus 2 (DENV2) in infected patients’ sera and 60% of the DENV IgM-positive sera not detected by commercial NS1-based diagnostic kits. These data suggest a high-efficiency method for correctly folding rNS1 that maintains its structural and immunogenic properties. In addition, a detection method using the polyclonal antibody against correctly folded rNS1 seemed to be more sensitive and efficient for NS1 detection in serum, highlighting its usefulness for developing a high-sensitivity diagnostic kit.</abstract><cop>Kidlington</cop><pub>Elsevier B.V</pub><pmid>21569796</pmid><doi>10.1016/j.jviromet.2011.04.029</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0166-0934 |
| ispartof | Journal of virological methods, 2011-07, Vol.175 (1), p.109-116 |
| issn | 0166-0934 1879-0984 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_907166544 |
| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Animals Antibodies, Viral - blood Antibodies, Viral - immunology Biological and medical sciences Blotting, Western Dengue - diagnosis Dengue virus Dengue Virus - genetics Dengue Virus - immunology Dengue virus type 2 E. coli Early Diagnosis Enzyme-Linked Immunosorbent Assay - methods Escherichia coli Escherichia coli - genetics Escherichia coli - metabolism Fundamental and applied biological sciences. Psychology Glycoproteins - immunology Humans Mice Mice, Inbred BALB C Microbiology NS1 protein Protein Folding Rabbits Techniques used in virology Viral Nonstructural Proteins - genetics Viral Nonstructural Proteins - immunology Viral Nonstructural Proteins - metabolism Virology |
| title | Polyclonal antibodies against properly folded Dengue virus NS1 protein expressed in E. coli enable sensitive and early dengue diagnosis |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-28T12%3A29%3A23IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Polyclonal%20antibodies%20against%20properly%20folded%20Dengue%20virus%20NS1%20protein%20expressed%20in%20E.%20coli%20enable%20sensitive%20and%20early%20dengue%20diagnosis&rft.jtitle=Journal%20of%20virological%20methods&rft.au=Allonso,%20Diego&rft.date=2011-07-01&rft.volume=175&rft.issue=1&rft.spage=109&rft.epage=116&rft.pages=109-116&rft.issn=0166-0934&rft.eissn=1879-0984&rft.coden=JVMEDH&rft_id=info:doi/10.1016/j.jviromet.2011.04.029&rft_dat=%3Cproquest_cross%3E871004255%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=871004255&rft_id=info:pmid/21569796&rft_els_id=S0166093411001674&rfr_iscdi=true |