Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus
Identifieur interne : 005B49 ( Main/Exploration ); précédent : 005B48; suivant : 005B50Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus
Auteurs : Jody D. Berry [Canada] ; Steven Jones [Canada] ; Michael A. Drebot [Canada] ; Anton Andonov [Canada] ; Marta Sabara [Canada] ; Xin Y. Yuan [Canada] ; Hana Weingartl [Canada] ; Lisa Fernando [Canada] ; Peter Marszal [Canada] ; Jason Gren [Canada] ; Brigitte Nicolas [Canada] ; Maya Andonova [Canada] ; Francesca Ranada [Canada] ; Michael J. Gubbins [Canada] ; T. Blake Ball [Canada] ; Paul Kitching [Canada] ; Yan Li [Canada] ; Amin Kabani [Canada] ; Frank Plummer [Canada]Source :
- Journal of virological methods [ 0166-0934 ] ; 2004.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (immunologie), Anticorps monoclonaux (immunologie), Antigènes viraux (immunologie), Cellules Vero, Conformation des protéines, Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (immunologie), Immunoglobuline G (analyse), Immunoglobuline G (immunologie), Nucléoprotéines (immunologie), Protéines de l'enveloppe virale (immunologie), Souris, Technique de Western, Test ELISA, Tests de neutralisation, Virus du SRAS (immunologie), Épitopes.
- MESH :
- Pascal (Inist)
- Animaux, Cellules Vero, Conformation des protéines, Coronavirus, Développement, Anticorps monoclonal, Déterminant antigénique, Glycoprotéine de spicule des coronavirus, Immunochimie, Microbiologie, Méthode, Souris, Technique de Western, Test ELISA, Tests de neutralisation, Virologie, Syndrome respiratoire aigu sévère, Épitopes.
English descriptors
- KwdEn :
- Animals, Antibodies, Monoclonal (immunology), Antibodies, Viral (immunology), Antigenic determinant, Antigens, Viral (immunology), Blotting, Western, Chlorocebus aethiops, Coronavirus, Development, Enzyme-Linked Immunosorbent Assay, Epitopes, Immunochemistry, Immunoglobulin G (analysis), Immunoglobulin G (immunology), Membrane Glycoproteins (immunology), Method, Mice, Microbiology, Monoclonal antibody, Neutralization Tests, Nucleoproteins (immunology), Protein Conformation, SARS Virus (immunology), Severe acute respiratory syndrome, Spike Glycoprotein, Coronavirus, Vero Cells, Viral Envelope Proteins (immunology), Virology.
- MESH :
- chemical , analysis : Immunoglobulin G.
- chemical , immunology : Antibodies, Monoclonal, Antibodies, Viral, Antigens, Viral, Immunoglobulin G, Membrane Glycoproteins, Nucleoproteins, Viral Envelope Proteins.
- immunology : SARS Virus.
- Animals, Blotting, Western, Chlorocebus aethiops, Enzyme-Linked Immunosorbent Assay, Epitopes, Mice, Neutralization Tests, Protein Conformation, Spike Glycoprotein, Coronavirus, Vero Cells.
Abstract
There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report, the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented, based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb, F26G15, is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients, but not normal human serum, is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests, studies of SARS-CoV pathogenesis and vaccine development.
Affiliations:
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Le document en format XML
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Blake Ball</name><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Medical Microbiology, University of Manitoba</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>15 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation></author><author><name sortKey="Kitching, Paul" sort="Kitching, Paul" uniqKey="Kitching P" first="Paul" last="Kitching">Paul Kitching</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 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type="wicri:Area/Ncbi/Curation">000933</idno><idno type="wicri:Area/Ncbi/Checkpoint">000933</idno><idno type="wicri:doubleKey">0166-0934:2004:Berry J:development:and:characterisation</idno><idno type="wicri:Area/Main/Merge">005813</idno><idno type="wicri:Area/Main/Curation">005B49</idno><idno type="wicri:Area/Main/Exploration">005B49</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus</title><author><name sortKey="Berry, Jody D" sort="Berry, Jody D" uniqKey="Berry J" first="Jody D." last="Berry">Jody D. Berry</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Winnipeg</wicri:noRegion></affiliation><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Medical Microbiology, University of Manitoba</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>15 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation></author><author><name sortKey="Jones, Steven" sort="Jones, Steven" uniqKey="Jones S" first="Steven" last="Jones">Steven Jones</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation><affiliation wicri:level="4"><inist:fA14 i1="04"><s1>Department of Immunology, University of Manitoba</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation></author><author><name sortKey="Drebot, Michael A" sort="Drebot, Michael A" uniqKey="Drebot M" first="Michael A." last="Drebot">Michael A. Drebot</name><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Medical Microbiology, University of Manitoba</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>15 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Andonov, Anton" sort="Andonov, Anton" uniqKey="Andonov A" first="Anton" last="Andonov">Anton Andonov</name><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Medical Microbiology, University of Manitoba</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>15 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Sabara, Marta" sort="Sabara, Marta" uniqKey="Sabara M" first="Marta" last="Sabara">Marta Sabara</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Winnipeg</wicri:noRegion></affiliation></author><author><name sortKey="Yuan, Xin Y" sort="Yuan, Xin Y" uniqKey="Yuan X" first="Xin Y." last="Yuan">Xin Y. Yuan</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Winnipeg</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Weingartl, Hana" sort="Weingartl, Hana" uniqKey="Weingartl H" first="Hana" last="Weingartl">Hana Weingartl</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Winnipeg</wicri:noRegion></affiliation></author><author><name sortKey="Fernando, Lisa" sort="Fernando, Lisa" uniqKey="Fernando L" first="Lisa" last="Fernando">Lisa Fernando</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Marszal, Peter" sort="Marszal, Peter" uniqKey="Marszal P" first="Peter" last="Marszal">Peter Marszal</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Winnipeg</wicri:noRegion></affiliation></author><author><name sortKey="Gren, Jason" sort="Gren, Jason" uniqKey="Gren J" first="Jason" last="Gren">Jason Gren</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Winnipeg</wicri:noRegion></affiliation></author><author><name sortKey="Nicolas, Brigitte" sort="Nicolas, Brigitte" uniqKey="Nicolas B" first="Brigitte" last="Nicolas">Brigitte Nicolas</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Winnipeg</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Andonova, Maya" sort="Andonova, Maya" uniqKey="Andonova M" first="Maya" last="Andonova">Maya Andonova</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Ranada, Francesca" sort="Ranada, Francesca" uniqKey="Ranada F" first="Francesca" last="Ranada">Francesca Ranada</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Winnipeg</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Gubbins, Michael J" sort="Gubbins, Michael J" uniqKey="Gubbins M" first="Michael J." last="Gubbins">Michael J. Gubbins</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Ball, T Blake" sort="Ball, T Blake" uniqKey="Ball T" first="T. Blake" last="Ball">T. Blake Ball</name><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Medical Microbiology, University of Manitoba</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>15 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation></author><author><name sortKey="Kitching, Paul" sort="Kitching, Paul" uniqKey="Kitching P" first="Paul" last="Kitching">Paul Kitching</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>National Centre for Foreign Animal Disease, CFIA</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ><sZ>11 aut.</sZ><sZ>13 aut.</sZ><sZ>16 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>Winnipeg</wicri:noRegion></affiliation></author><author><name sortKey="Li, Yan" sort="Li, Yan" uniqKey="Li Y" first="Yan" last="Li">Yan Li</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Kabani, Amin" sort="Kabani, Amin" uniqKey="Kabani A" first="Amin" last="Kabani">Amin Kabani</name><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Medical Microbiology, University of Manitoba</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>15 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></affiliation></author><author><name sortKey="Plummer, Frank" sort="Plummer, Frank" uniqKey="Plummer F" first="Frank" last="Plummer">Frank Plummer</name><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Medical Microbiology, University of Manitoba</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>15 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><placeName><settlement type="city">Winnipeg</settlement><region type="state">Manitoba</region></placeName><orgName type="university">Université du Manitoba</orgName></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>National Microbiology Laboratory Health Canada</s1><s2>Winnipeg</s2><s3>CAN</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>11 aut.</sZ><sZ>12 aut.</sZ><sZ>13 aut.</sZ><sZ>14 aut.</sZ><sZ>17 aut.</sZ><sZ>18 aut.</sZ><sZ>19 aut.</sZ></inist:fA14><country>Canada</country><wicri:noRegion>National Microbiology Laboratory Health Canada</wicri:noRegion></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="2004">2004</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>Animals</term><term>Antibodies, Monoclonal (immunology)</term><term>Antibodies, Viral (immunology)</term><term>Antigenic determinant</term><term>Antigens, Viral (immunology)</term><term>Blotting, Western</term><term>Chlorocebus aethiops</term><term>Coronavirus</term><term>Development</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Epitopes</term><term>Immunochemistry</term><term>Immunoglobulin G (analysis)</term><term>Immunoglobulin G (immunology)</term><term>Membrane Glycoproteins (immunology)</term><term>Method</term><term>Mice</term><term>Microbiology</term><term>Monoclonal antibody</term><term>Neutralization Tests</term><term>Nucleoproteins (immunology)</term><term>Protein Conformation</term><term>SARS Virus (immunology)</term><term>Severe acute respiratory syndrome</term><term>Spike Glycoprotein, Coronavirus</term><term>Vero Cells</term><term>Viral Envelope Proteins (immunology)</term><term>Virology</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps antiviraux (immunologie)</term><term>Anticorps monoclonaux (immunologie)</term><term>Antigènes viraux (immunologie)</term><term>Cellules Vero</term><term>Conformation des protéines</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (immunologie)</term><term>Immunoglobuline G (analyse)</term><term>Immunoglobuline G (immunologie)</term><term>Nucléoprotéines (immunologie)</term><term>Protéines de l'enveloppe virale (immunologie)</term><term>Souris</term><term>Technique de Western</term><term>Test ELISA</term><term>Tests de neutralisation</term><term>Virus du SRAS (immunologie)</term><term>Épitopes</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Immunoglobulin G</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Monoclonal</term><term>Antibodies, Viral</term><term>Antigens, Viral</term><term>Immunoglobulin G</term><term>Membrane Glycoproteins</term><term>Nucleoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Immunoglobuline G</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps monoclonaux</term><term>Antigènes viraux</term><term>Glycoprotéines membranaires</term><term>Immunoglobuline G</term><term>Nucléoprotéines</term><term>Protéines de l'enveloppe virale</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Blotting, Western</term><term>Chlorocebus aethiops</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Epitopes</term><term>Mice</term><term>Neutralization Tests</term><term>Protein Conformation</term><term>Spike Glycoprotein, Coronavirus</term><term>Vero Cells</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>Conformation des protéines</term><term>Coronavirus</term><term>Développement</term><term>Anticorps monoclonal</term><term>Déterminant antigénique</term><term>Glycoprotéine de spicule des coronavirus</term><term>Immunochimie</term><term>Microbiologie</term><term>Méthode</term><term>Souris</term><term>Technique de Western</term><term>Test ELISA</term><term>Tests de neutralisation</term><term>Virologie</term><term>Syndrome respiratoire aigu sévère</term><term>Épitopes</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report, the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented, based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb, F26G15, is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients, but not normal human serum, is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests, studies of SARS-CoV pathogenesis and vaccine development.</div></front></TEI><affiliations><list><country><li>Canada</li></country><region><li>Manitoba</li></region><settlement><li>Winnipeg</li></settlement><orgName><li>Université du Manitoba</li></orgName></list><tree><country name="Canada"><noRegion><name sortKey="Berry, Jody D" sort="Berry, Jody D" uniqKey="Berry J" first="Jody D." last="Berry">Jody D. Berry</name></noRegion><name sortKey="Andonov, Anton" sort="Andonov, Anton" uniqKey="Andonov A" first="Anton" last="Andonov">Anton Andonov</name><name sortKey="Andonov, Anton" sort="Andonov, Anton" uniqKey="Andonov A" first="Anton" last="Andonov">Anton Andonov</name><name sortKey="Andonova, Maya" sort="Andonova, Maya" uniqKey="Andonova M" first="Maya" last="Andonova">Maya Andonova</name><name sortKey="Ball, T Blake" sort="Ball, T Blake" uniqKey="Ball T" first="T. Blake" last="Ball">T. Blake Ball</name><name sortKey="Berry, Jody D" sort="Berry, Jody D" uniqKey="Berry J" first="Jody D." last="Berry">Jody D. Berry</name><name sortKey="Drebot, Michael A" sort="Drebot, Michael A" uniqKey="Drebot M" first="Michael A." last="Drebot">Michael A. Drebot</name><name sortKey="Drebot, Michael A" sort="Drebot, Michael A" uniqKey="Drebot M" first="Michael A." last="Drebot">Michael A. Drebot</name><name sortKey="Fernando, Lisa" sort="Fernando, Lisa" uniqKey="Fernando L" first="Lisa" last="Fernando">Lisa Fernando</name><name sortKey="Gren, Jason" sort="Gren, Jason" uniqKey="Gren J" first="Jason" last="Gren">Jason Gren</name><name sortKey="Gubbins, Michael J" sort="Gubbins, Michael J" uniqKey="Gubbins M" first="Michael J." last="Gubbins">Michael J. Gubbins</name><name sortKey="Jones, Steven" sort="Jones, Steven" uniqKey="Jones S" first="Steven" last="Jones">Steven Jones</name><name sortKey="Jones, Steven" sort="Jones, Steven" uniqKey="Jones S" first="Steven" last="Jones">Steven Jones</name><name sortKey="Kabani, Amin" sort="Kabani, Amin" uniqKey="Kabani A" first="Amin" last="Kabani">Amin Kabani</name><name sortKey="Kabani, Amin" sort="Kabani, Amin" uniqKey="Kabani A" first="Amin" last="Kabani">Amin Kabani</name><name sortKey="Kitching, Paul" sort="Kitching, Paul" uniqKey="Kitching P" first="Paul" last="Kitching">Paul Kitching</name><name sortKey="Li, Yan" sort="Li, Yan" uniqKey="Li Y" first="Yan" last="Li">Yan Li</name><name sortKey="Marszal, Peter" sort="Marszal, Peter" uniqKey="Marszal P" first="Peter" last="Marszal">Peter Marszal</name><name sortKey="Nicolas, Brigitte" sort="Nicolas, Brigitte" uniqKey="Nicolas B" first="Brigitte" last="Nicolas">Brigitte Nicolas</name><name sortKey="Nicolas, Brigitte" sort="Nicolas, Brigitte" uniqKey="Nicolas B" first="Brigitte" last="Nicolas">Brigitte Nicolas</name><name sortKey="Plummer, Frank" sort="Plummer, Frank" uniqKey="Plummer F" first="Frank" last="Plummer">Frank Plummer</name><name sortKey="Plummer, Frank" sort="Plummer, Frank" uniqKey="Plummer F" first="Frank" last="Plummer">Frank Plummer</name><name sortKey="Ranada, Francesca" sort="Ranada, Francesca" uniqKey="Ranada F" first="Francesca" last="Ranada">Francesca Ranada</name><name sortKey="Ranada, Francesca" sort="Ranada, Francesca" uniqKey="Ranada F" first="Francesca" last="Ranada">Francesca Ranada</name><name sortKey="Sabara, Marta" sort="Sabara, Marta" uniqKey="Sabara M" first="Marta" last="Sabara">Marta Sabara</name><name sortKey="Weingartl, Hana" sort="Weingartl, Hana" uniqKey="Weingartl H" first="Hana" last="Weingartl">Hana Weingartl</name><name sortKey="Yuan, Xin Y" sort="Yuan, Xin Y" uniqKey="Yuan X" first="Xin Y." last="Yuan">Xin Y. Yuan</name><name sortKey="Yuan, Xin Y" sort="Yuan, Xin Y" uniqKey="Yuan X" first="Xin Y." last="Yuan">Xin Y. Yuan</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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