Characterization of monoclonal antibody against SARS coronavirus nucleocapsid antigen and development of an antigen capture ELISA
Identifieur interne : 004E62 ( Main/Curation ); précédent : 004E61; suivant : 004E63Characterization of monoclonal antibody against SARS coronavirus nucleocapsid antigen and development of an antigen capture ELISA
Auteurs : QIGAI HE [Singapour] ; QINGYUN DU [Singapour] ; Suelyn Lau [Singapour] ; Ivanus Manopo [Singapour] ; LIQUN LU [Singapour] ; Shzu-Wei Chan [Singapour] ; Beau J. Fenner [Singapour] ; Jimmy Kwang [Singapour]Source :
- Journal of virological methods [ 0166-0934 ] ; 2005.
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Abstract
This report describes the production of several MAbs against N195 protein, a major immunodomain of SARS CoV nucleocapsid protein [He, Q., Chong, K.H.. Chang, H.H., Leung, B., Ling, A.E., Wei, T., Chan, S.W., Ooi, E.E., Kwang, J., 2004. Development of a Western blot assay for detection of antibodies against coronavirus causing severe acute respiratory syndrome. Clin. Diagn. Lab. Immunol. 11 (2)417-422.]. One representative IgG1 monoclonal antibody (MAb), S-A5D5, was selected and characterized. S-A5D5 reacted specifically react with both recombinant and native nucleocapsid protein of SARS CoV. The reactivity of S-A5D5 with purified N195 protein and utilization of the MAb as a detector antibody to develop an antigen capture ELISA was assessed. As little as 37.5 pg of purified N protein and 50 TCID50 of SARS CoV could be detected by the antigen capture ELISA. Specific binding of the MAb S-A5D5 to both purified N195 and SARS CoV nucleocapsid antigen was effectively inhibited by human SARS positive serum and guinea pig anti-N195 serum. The N protein in N195-spike recombinant baculovirus-infected Sf-9 cells could also be identified. N protein was detected in 18 IFA IgM-positive serum samples collected from SARS confirmed patients, but not in nine samples collected from SARS recovery patient. No false positive results were given when 60 samples from healthy individuals were tested, and no cross-reaction occurred when infectious bronchitis virus (IBV), chicken coronavirus, was tested. This monoclonal antibody-based antigen capture ELISA is thus a powerful tool for early diagnosis of SARS CoV infection.
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Fenner</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author><author><name sortKey="Kwang, Jimmy" sort="Kwang, Jimmy" uniqKey="Kwang J" first="Jimmy" last="Kwang">Jimmy Kwang</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">05-0260154</idno><date when="2005">2005</date><idno type="stanalyst">PASCAL 05-0260154 INIST</idno><idno type="RBID">Pascal:05-0260154</idno><idno type="wicri:Area/PascalFrancis/Corpus">000668</idno><idno type="wicri:Area/PascalFrancis/Curation">000322</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000692</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000692</idno><idno type="wicri:doubleKey">0166-0934:2005:Qigai He:characterization:of:monoclonal</idno><idno type="wicri:Area/Main/Merge">005212</idno><idno type="wicri:Area/Main/Curation">004E62</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Characterization of monoclonal antibody against SARS coronavirus nucleocapsid antigen and development of an antigen capture ELISA</title><author><name sortKey="Qigai He" sort="Qigai He" uniqKey="Qigai He" last="Qigai He">QIGAI HE</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author><author><name sortKey="Qingyun Du" sort="Qingyun Du" uniqKey="Qingyun Du" last="Qingyun Du">QINGYUN DU</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author><author><name sortKey="Lau, Suelyn" sort="Lau, Suelyn" uniqKey="Lau S" first="Suelyn" last="Lau">Suelyn Lau</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author><author><name sortKey="Manopo, Ivanus" sort="Manopo, Ivanus" uniqKey="Manopo I" first="Ivanus" last="Manopo">Ivanus Manopo</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author><author><name sortKey="Liqun Lu" sort="Liqun Lu" uniqKey="Liqun Lu" last="Liqun Lu">LIQUN LU</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author><author><name sortKey="Chan, Shzu Wei" sort="Chan, Shzu Wei" uniqKey="Chan S" first="Shzu-Wei" last="Chan">Shzu-Wei Chan</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author><author><name sortKey="Fenner, Beau J" sort="Fenner, Beau J" uniqKey="Fenner B" first="Beau J." last="Fenner">Beau J. Fenner</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author><author><name sortKey="Kwang, Jimmy" sort="Kwang, Jimmy" uniqKey="Kwang J" first="Jimmy" last="Kwang">Jimmy Kwang</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Animal Health Biotechnology, Temasek Life Science Laboratory, I Research Link, National University of Singapore</s1><s2>Singapore 117604</s2><s3>SGP</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Singapour</country><orgName type="university">Université nationale de Singapour</orgName></affiliation></author></analytic><series><title level="j" type="main">Journal of virological methods</title><title level="j" type="abbreviated">J. virol. methods</title><idno type="ISSN">0166-0934</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of virological methods</title><title level="j" type="abbreviated">J. virol. methods</title><idno type="ISSN">0166-0934</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antigen</term><term>ELISA assay</term><term>Method</term><term>Microbiology</term><term>Monoclonal antibody</term><term>Nucleocapsid</term><term>Severe acute respiratory syndrome</term><term>Severe acute respiratory syndrome virus</term><term>Virology</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Virus syndrome respiratoire aigu sévère</term><term>Anticorps monoclonal</term><term>Nucléocapside</term><term>Antigène</term><term>Technique ELISA</term><term>Microbiologie</term><term>Méthode</term><term>Virologie</term><term>Syndrome respiratoire aigu sévère</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">This report describes the production of several MAbs against N195 protein, a major immunodomain of SARS CoV nucleocapsid protein [He, Q., Chong, K.H.. Chang, H.H., Leung, B., Ling, A.E., Wei, T., Chan, S.W., Ooi, E.E., Kwang, J., 2004. Development of a Western blot assay for detection of antibodies against coronavirus causing severe acute respiratory syndrome. Clin. Diagn. Lab. Immunol. 11 (2)417-422.]. One representative IgG1 monoclonal antibody (MAb), S-A5D5, was selected and characterized. S-A5D5 reacted specifically react with both recombinant and native nucleocapsid protein of SARS CoV. The reactivity of S-A5D5 with purified N195 protein and utilization of the MAb as a detector antibody to develop an antigen capture ELISA was assessed. As little as 37.5 pg of purified N protein and 50 TCID<sub>50</sub> of SARS CoV could be detected by the antigen capture ELISA. Specific binding of the MAb S-A5D5 to both purified N195 and SARS CoV nucleocapsid antigen was effectively inhibited by human SARS positive serum and guinea pig anti-N195 serum. The N protein in N195-spike recombinant baculovirus-infected Sf-9 cells could also be identified. N protein was detected in 18 IFA IgM-positive serum samples collected from SARS confirmed patients, but not in nine samples collected from SARS recovery patient. No false positive results were given when 60 samples from healthy individuals were tested, and no cross-reaction occurred when infectious bronchitis virus (IBV), chicken coronavirus, was tested. This monoclonal antibody-based antigen capture ELISA is thus a powerful tool for early diagnosis of SARS CoV infection.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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