Generation of human antibody fragments recognizing distinct epitopes of the nucleocapsid (N) SARS-CoV protein using a phage display approach
Identifieur interne : 000289 ( Pmc/Curation ); précédent : 000288; suivant : 000290Generation of human antibody fragments recognizing distinct epitopes of the nucleocapsid (N) SARS-CoV protein using a phage display approach
Auteurs : Michela Flego [Italie] ; Paola Di Bonito [Italie] ; Alessandro Ascione [Italie] ; Silvia Zamboni [Italie] ; Alessandra Carattoli [Italie] ; Felicia Grasso [Italie] ; Antonio Cassone [Italie] ; Maurizio Cianfriglia [Italie]Source :
- BMC Infectious Diseases [ 1471-2334 ] ; 2005.
Abstract
Severe acute respiratory syndrome (SARS)-CoV is a newly emerging virus that causes SARS with high mortality rate in infected people. Successful control of the global SARS epidemic will require rapid and sensitive diagnostic tests to monitor its spread, as well as, the development of vaccines and new antiviral compounds including neutralizing antibodies that effectively prevent or treat this disease.
The human synthetic single-chain fragment variable (scFv) ETH-2 phage antibody library was used for the isolation of scFvs against the nucleocapsid (N) protein of SARS-CoV using a bio panning-based strategy. The selected scFvs were characterized under genetics-molecular aspects and for SARS-CoV N protein detection in ELISA, western blotting and immunocytochemistry.
Human scFv antibodies to N protein of SARS-CoV can be easily isolated by selecting the ETH-2 phage library on immunotubes coated with antigen. These
The human scFv antibodies isolated and described in this study represent useful reagents for rapid detection of N SARS-CoV protein and SARS virus particles in infected target cells.
The online version of this article (doi:10.1186/1471-2334-5-73) contains supplementary material, which is available to authorized users.
Url:
DOI: 10.1186/1471-2334-5-73
PubMed: 16171519
PubMed Central: 1261265
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wicri:stream="Pmc" wicri:step="Curation">000289</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Generation of human antibody fragments recognizing distinct epitopes of the nucleocapsid (N) SARS-CoV protein using a phage display approach</title><author><name sortKey="Flego, Michela" sort="Flego, Michela" uniqKey="Flego M" first="Michela" last="Flego">Michela Flego</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.416651.1</institution-id><institution-id institution-id-type="ISNI">0000000091206856</institution-id><institution>Department of Drug Research and Evaluation,</institution><institution>Istituto Superiore di Sanità,</institution></institution-wrap>Rome, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Rome</wicri:regionArea></affiliation></author><author><name sortKey="Di Bonito, Paola" sort="Di Bonito, Paola" uniqKey="Di Bonito P" first="Paola" last="Di Bonito">Paola Di Bonito</name><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.416651.1</institution-id><institution-id institution-id-type="ISNI">0000000091206856</institution-id><institution>Department of Infectious Parasitic and Immune-mediated Diseases,</institution><institution>Istituto Superiore di Sanità,</institution></institution-wrap>Rome, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Rome</wicri:regionArea></affiliation></author><author><name sortKey="Ascione, Alessandro" sort="Ascione, Alessandro" uniqKey="Ascione A" first="Alessandro" last="Ascione">Alessandro Ascione</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.416651.1</institution-id><institution-id institution-id-type="ISNI">0000000091206856</institution-id><institution>Department of Drug Research and 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Carattoli</name><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.416651.1</institution-id><institution-id institution-id-type="ISNI">0000000091206856</institution-id><institution>Department of Infectious Parasitic and Immune-mediated Diseases,</institution><institution>Istituto Superiore di Sanità,</institution></institution-wrap>Rome, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Rome</wicri:regionArea></affiliation></author><author><name sortKey="Grasso, Felicia" sort="Grasso, Felicia" uniqKey="Grasso F" first="Felicia" last="Grasso">Felicia Grasso</name><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.416651.1</institution-id><institution-id institution-id-type="ISNI">0000000091206856</institution-id><institution>Department of Infectious Parasitic and Immune-mediated Diseases,</institution><institution>Istituto Superiore di Sanità,</institution></institution-wrap>Rome, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Rome</wicri:regionArea></affiliation></author><author><name sortKey="Cassone, Antonio" sort="Cassone, Antonio" uniqKey="Cassone A" first="Antonio" last="Cassone">Antonio Cassone</name><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="GRID">grid.416651.1</institution-id><institution-id institution-id-type="ISNI">0000000091206856</institution-id><institution>Department of Infectious Parasitic and Immune-mediated Diseases,</institution><institution>Istituto Superiore di Sanità,</institution></institution-wrap>Rome, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Rome</wicri:regionArea></affiliation></author><author><name sortKey="Cianfriglia, Maurizio" sort="Cianfriglia, Maurizio" uniqKey="Cianfriglia M" first="Maurizio" last="Cianfriglia">Maurizio Cianfriglia</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="GRID">grid.416651.1</institution-id><institution-id institution-id-type="ISNI">0000000091206856</institution-id><institution>Department of Drug Research and Evaluation,</institution><institution>Istituto Superiore di Sanità,</institution></institution-wrap>Rome, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Rome</wicri:regionArea></affiliation></author></analytic><series><title level="j">BMC Infectious Diseases</title><idno type="eISSN">1471-2334</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>Severe acute respiratory syndrome (SARS)-CoV is a newly emerging virus that causes SARS with high mortality rate in infected people. Successful control of the global SARS epidemic will require rapid and sensitive diagnostic tests to monitor its spread, as well as, the development of vaccines and new antiviral compounds including neutralizing antibodies that effectively prevent or treat this disease.</p></sec><sec><title>Methods</title><p>The human synthetic single-chain fragment variable (scFv) ETH-2 phage antibody library was used for the isolation of scFvs against the nucleocapsid (N) protein of SARS-CoV using a bio panning-based strategy. The selected scFvs were characterized under genetics-molecular aspects and for SARS-CoV N protein detection in ELISA, western blotting and immunocytochemistry.</p></sec><sec><title>Results</title><p>Human scFv antibodies to N protein of SARS-CoV can be easily isolated by selecting the ETH-2 phage library on immunotubes coated with antigen. These <italic>in vitro</italic> selected human scFvs specifically recognize in ELISA and western blotting studies distinct epitopes in N protein domains and detect in immunohistochemistry investigations SARS-CoV particles in infected Vero cells.</p></sec><sec><title>Conclusion</title><p>The human scFv antibodies isolated and described in this study represent useful reagents for rapid detection of N SARS-CoV protein and SARS virus particles in infected target cells.</p></sec><sec><title>Electronic supplementary material</title><p>The online version of this article (doi:10.1186/1471-2334-5-73) contains supplementary material, which is available to authorized users.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Bermingham, A" uniqKey="Bermingham A">A Bermingham</name></author><author><name sortKey="Heinen, P" uniqKey="Heinen P">P Heinen</name></author><author><name sortKey="Iturriza Gomara, M" 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<front><journal-meta><journal-id journal-id-type="nlm-ta">BMC Infect Dis</journal-id><journal-id journal-id-type="iso-abbrev">BMC Infect. Dis</journal-id><journal-title-group><journal-title>BMC Infectious Diseases</journal-title></journal-title-group><issn pub-type="epub">1471-2334</issn><publisher><publisher-name>BioMed Central</publisher-name><publisher-loc>London</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">16171519</article-id><article-id pub-id-type="pmc">1261265</article-id><article-id pub-id-type="publisher-id">221</article-id><article-id pub-id-type="doi">10.1186/1471-2334-5-73</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title>Generation of human antibody fragments recognizing distinct epitopes of the nucleocapsid (N) SARS-CoV protein using a phage display approach</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Flego</surname><given-names>Michela</given-names></name><address><email>flego@iss.it</email></address><xref ref-type="aff" rid="Aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Di Bonito</surname><given-names>Paola</given-names></name><address><email>dibonito@iss.it</email></address><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Ascione</surname><given-names>Alessandro</given-names></name><address><email>fiano1974@Yahoo.it</email></address><xref ref-type="aff" rid="Aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Zamboni</surname><given-names>Silvia</given-names></name><address><email>zamboni@iss.it</email></address><xref ref-type="aff" rid="Aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Carattoli</surname><given-names>Alessandra</given-names></name><address><email>alecara@iss.it</email></address><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Grasso</surname><given-names>Felicia</given-names></name><address><email>grasso@iss.it</email></address><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Cassone</surname><given-names>Antonio</given-names></name><address><email>cassone@iss.it</email></address><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Cianfriglia</surname><given-names>Maurizio</given-names></name><address><email>cianfri@iss.it</email></address><xref ref-type="aff" rid="Aff1">1</xref></contrib><aff id="Aff1"><label>1</label><institution-wrap><institution-id institution-id-type="GRID">grid.416651.1</institution-id><institution-id institution-id-type="ISNI">0000000091206856</institution-id><institution>Department of Drug Research and Evaluation,</institution><institution>Istituto Superiore di Sanità,</institution></institution-wrap>Rome, Italy</aff><aff id="Aff2"><label>2</label><institution-wrap><institution-id institution-id-type="GRID">grid.416651.1</institution-id><institution-id institution-id-type="ISNI">0000000091206856</institution-id><institution>Department of Infectious Parasitic and Immune-mediated Diseases,</institution><institution>Istituto Superiore di Sanità,</institution></institution-wrap>Rome, Italy</aff></contrib-group><pub-date pub-type="epub"><day>19</day><month>9</month><year>2005</year></pub-date><pub-date pub-type="pmc-release"><day>19</day><month>9</month><year>2005</year></pub-date><pub-date pub-type="collection"><year>2005</year></pub-date><volume>5</volume><elocation-id>73</elocation-id><history><date date-type="received"><day>18</day><month>4</month><year>2005</year></date><date date-type="accepted"><day>19</day><month>9</month><year>2005</year></date></history><permissions><copyright-statement>© Flego et al; licensee BioMed Central Ltd. 2005</copyright-statement><license><license-p>This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.</license-p></license></permissions><abstract id="Abs1"><sec><title>Background</title><p>Severe acute respiratory syndrome (SARS)-CoV is a newly emerging virus that causes SARS with high mortality rate in infected people. Successful control of the global SARS epidemic will require rapid and sensitive diagnostic tests to monitor its spread, as well as, the development of vaccines and new antiviral compounds including neutralizing antibodies that effectively prevent or treat this disease.</p></sec><sec><title>Methods</title><p>The human synthetic single-chain fragment variable (scFv) ETH-2 phage antibody library was used for the isolation of scFvs against the nucleocapsid (N) protein of SARS-CoV using a bio panning-based strategy. The selected scFvs were characterized under genetics-molecular aspects and for SARS-CoV N protein detection in ELISA, western blotting and immunocytochemistry.</p></sec><sec><title>Results</title><p>Human scFv antibodies to N protein of SARS-CoV can be easily isolated by selecting the ETH-2 phage library on immunotubes coated with antigen. These <italic>in vitro</italic> selected human scFvs specifically recognize in ELISA and western blotting studies distinct epitopes in N protein domains and detect in immunohistochemistry investigations SARS-CoV particles in infected Vero cells.</p></sec><sec><title>Conclusion</title><p>The human scFv antibodies isolated and described in this study represent useful reagents for rapid detection of N SARS-CoV protein and SARS virus particles in infected target cells.</p></sec><sec><title>Electronic supplementary material</title><p>The online version of this article (doi:10.1186/1471-2334-5-73) contains supplementary material, which is available to authorized users.</p></sec></abstract><kwd-group xml:lang="en"><title>Keywords</title><kwd>Severe Acute Respiratory Syndrome</kwd><kwd>Severe Acute Respiratory Syndrome</kwd><kwd>scFv Antibody</kwd><kwd>Severe Acute Respiratory Syndrome Patient</kwd><kwd>Phage Antibody</kwd></kwd-group><custom-meta-group><custom-meta><meta-name>issue-copyright-statement</meta-name><meta-value>© The Author(s) 2005</meta-value></custom-meta></custom-meta-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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