Characterization of monoclonal antibody against SARS coronavirus nucleocapsid antigen and development of an antigen capture ELISA.
Identifieur interne : 000F49 ( Ncbi/Curation ); précédent : 000F48; suivant : 000F50Characterization of monoclonal antibody against SARS coronavirus nucleocapsid antigen and development of an antigen capture ELISA.
Auteurs : Qigai He [Singapour] ; Qingyun Du ; Suelyn Lau ; Ivanus Manopo ; Liqun Lu ; Shzu-Wei Chan ; Beau J. Fenner ; Jimmy KwangSource :
- Journal of virological methods [ 0166-0934 ] ; 2005.
Descripteurs français
- KwdFr :
- Anticorps antiviraux (immunologie), Anticorps monoclonaux (biosynthèse), Anticorps monoclonaux (immunologie), Antigènes viraux (analyse), Antigènes viraux (immunologie), Humains, Protéines nucléocapside (analyse), Protéines nucléocapside (immunologie), Sensibilité et spécificité, Spécificité des anticorps, Test ELISA (), Virus du SRAS (immunologie), Virus du SRAS (isolement et purification).
- MESH :
- analyse : Antigènes viraux, Protéines nucléocapside.
- biosynthèse : Anticorps monoclonaux.
- immunologie : Anticorps antiviraux, Anticorps monoclonaux, Antigènes viraux, Protéines nucléocapside, Virus du SRAS.
- isolement et purification : Virus du SRAS.
- Humains, Sensibilité et spécificité, Spécificité des anticorps, Test ELISA.
English descriptors
- KwdEn :
- Antibodies, Monoclonal (biosynthesis), Antibodies, Monoclonal (immunology), Antibodies, Viral (immunology), Antibody Specificity, Antigens, Viral (analysis), Antigens, Viral (immunology), Enzyme-Linked Immunosorbent Assay (methods), Humans, Nucleocapsid Proteins (analysis), Nucleocapsid Proteins (immunology), SARS Virus (immunology), SARS Virus (isolation & purification), Sensitivity and Specificity.
- MESH :
- chemical , analysis : Antigens, Viral, Nucleocapsid Proteins.
- chemical , biosynthesis : Antibodies, Monoclonal.
- chemical , immunology : Antibodies, Monoclonal, Antibodies, Viral, Antigens, Viral, Nucleocapsid Proteins.
- immunology : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Enzyme-Linked Immunosorbent Assay.
- Antibody Specificity, Humans, Sensitivity and Specificity.
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 TCID(50) 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.
DOI: 10.1016/j.jviromet.2005.03.004
PubMed: 15893565
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Fenner</name></author><author><name sortKey="Kwang, Jimmy" sort="Kwang, Jimmy" uniqKey="Kwang J" first="Jimmy" last="Kwang">Jimmy Kwang</name></author></analytic><series><title level="j">Journal of virological methods</title><idno type="ISSN">0166-0934</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Monoclonal (biosynthesis)</term><term>Antibodies, Monoclonal (immunology)</term><term>Antibodies, Viral (immunology)</term><term>Antibody Specificity</term><term>Antigens, Viral (analysis)</term><term>Antigens, Viral (immunology)</term><term>Enzyme-Linked Immunosorbent Assay (methods)</term><term>Humans</term><term>Nucleocapsid Proteins (analysis)</term><term>Nucleocapsid Proteins (immunology)</term><term>SARS Virus (immunology)</term><term>SARS Virus (isolation & purification)</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Anticorps antiviraux (immunologie)</term><term>Anticorps monoclonaux (biosynthèse)</term><term>Anticorps monoclonaux (immunologie)</term><term>Antigènes viraux (analyse)</term><term>Antigènes viraux (immunologie)</term><term>Humains</term><term>Protéines nucléocapside (analyse)</term><term>Protéines nucléocapside (immunologie)</term><term>Sensibilité et spécificité</term><term>Spécificité des anticorps</term><term>Test ELISA ()</term><term>Virus du SRAS (immunologie)</term><term>Virus du SRAS (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Antigens, Viral</term><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Antibodies, Monoclonal</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>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Antigènes viraux</term><term>Protéines nucléocapside</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Anticorps monoclonaux</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term><term>Anticorps monoclonaux</term><term>Antigènes viraux</term><term>Protéines nucléocapside</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Antibody Specificity</term><term>Humans</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Humains</term><term>Sensibilité et spécificité</term><term>Spécificité des anticorps</term><term>Test ELISA</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(50) 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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