Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus.
Identifieur interne : 003150 ( Main/Exploration ); précédent : 003149; suivant : 003151Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus.
Auteurs : Hiroaki Kariwa [Japon] ; Hiroshi Noda ; Mina Nakauchi ; Mariko Ishizuka ; Kazuaki Hashiguchi ; Shingo Hashimoto ; Kentaro Yoshii ; Atsushi Asano ; Takashi Agui ; Hiroyuki Kogaki ; Yoshihiro Kurano ; Yoshiaki Uchida ; Nobuyuki Fujii ; Masahisa Okada ; Ikuo TakashimaSource :
- The Japanese journal of veterinary research [ 0047-1917 ] ; 2008.
Descripteurs français
- KwdFr :
- Acides aminés (), Animaux, Anticorps antiviraux (), Anticorps monoclonaux (), Anticorps monoclonaux (immunologie), Cartographie épitopique, Cellules Vero, Clonage moléculaire, Coronavirus humain 229E, Humains, Lapins, Protéines nucléocapside (), Protéines nucléocapside (génétique), Protéines nucléocapside (immunologie), RT-PCR (médecine vétérinaire), Réactions croisées, Souris, Spécificité des anticorps, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (médecine vétérinaire), Séquence d'acides aminés, Technique d'immunofluorescence indirecte (médecine vétérinaire), Technique de Western (médecine vétérinaire), Virus du SRAS (immunologie).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Protéines nucléocapside.
- immunologie : Anticorps monoclonaux, Protéines nucléocapside, Virus du SRAS.
- médecine vétérinaire : RT-PCR, Syndrome respiratoire aigu sévère, Technique d'immunofluorescence indirecte, Technique de Western.
- Acides aminés, Animaux, Anticorps antiviraux, Anticorps monoclonaux, Cartographie épitopique, Cellules Vero, Clonage moléculaire, Coronavirus humain 229E, Humains, Lapins, Protéines nucléocapside, Réactions croisées, Souris, Spécificité des anticorps, Séquence d'acides aminés.
English descriptors
- KwdEn :
- Amino Acid Sequence, Amino Acids (chemistry), Animals, Antibodies, Monoclonal (chemistry), Antibodies, Monoclonal (immunology), Antibodies, Viral (chemistry), Antibody Specificity, Blotting, Western (veterinary), Chlorocebus aethiops, Cloning, Molecular, Coronavirus 229E, Human, Cross Reactions, Epitope Mapping, Fluorescent Antibody Technique, Indirect (veterinary), Humans, Mice, Nucleocapsid Proteins (chemistry), Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (immunology), Rabbits, Reverse Transcriptase Polymerase Chain Reaction (veterinary), SARS Virus (immunology), Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (veterinary), Vero Cells.
- MESH :
- chemical , chemistry : Amino Acids, Antibodies, Monoclonal, Antibodies, Viral, Nucleocapsid Proteins.
- chemical , genetics : Nucleocapsid Proteins.
- chemical , immunology : Antibodies, Monoclonal, Nucleocapsid Proteins.
- diagnosis : Severe Acute Respiratory Syndrome.
- immunology : SARS Virus.
- veterinary : Blotting, Western, Fluorescent Antibody Technique, Indirect, Reverse Transcriptase Polymerase Chain Reaction, Severe Acute Respiratory Syndrome.
- Amino Acid Sequence, Animals, Antibody Specificity, Chlorocebus aethiops, Cloning, Molecular, Coronavirus 229E, Human, Cross Reactions, Epitope Mapping, Humans, Mice, Rabbits, Vero Cells.
Abstract
The sudden emergence of severe acute respiratory syndrome (SARS) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. As no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a SARS epidemic is the rapid diagnosis and isolation of SARS patients. To establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (NP) of SARS-coronavirus (SARS-CoV). On immunofluorescent antibody assay and Western blotting analysis, none of the monoclonal antibodies showed cross-reactivity to authentic and recombinant NPs of human coronavirus (HCoV) 229E strain. To determine the region on the NP molecule where the monoclonal antibodies bind, we generated four truncated recombinant NPs and analyzed the reactivity between monoclonal antibodies and truncated NPs. Two monoclonal antibodies reacted with a truncated NP covering from amino acid residues 111 to 230, and seven reacted with another truncated NP covering from amino acid residues 221 to 340. Epitope mapping analysis indicated that monoclonal antibody SN5-25 recognized the amino acid sequence Q(245)TVTKK(250) On SARS-NP. Within the epitope, Q245, T246, V247, K249, and K250 appeared to form an essential motif for monoclonal antibody SN5-25 to bind. The information about binding sites and epitopes of monoclonal antibodies may be useful for the development of new diagnostic methods for SARS and for analyzing the function of N protein of SARS-CoV.
PubMed: 18380153
Affiliations:
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Le document en format XML
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xml:lang="en">Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus.</title><author><name sortKey="Kariwa, Hiroaki" sort="Kariwa, Hiroaki" uniqKey="Kariwa H" first="Hiroaki" last="Kariwa">Hiroaki Kariwa</name><affiliation wicri:level="1"><nlm:affiliation>Laboratory of Public Health, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan. kariwa@vetemed.hokudai.ac.jp</nlm:affiliation><country xml:lang="fr">Japon</country><wicri:regionArea>Laboratory of Public Health, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818</wicri:regionArea><wicri:noRegion>Sapporo 060-0818</wicri:noRegion></affiliation></author><author><name sortKey="Noda, Hiroshi" sort="Noda, Hiroshi" uniqKey="Noda H" first="Hiroshi" last="Noda">Hiroshi Noda</name></author><author><name sortKey="Nakauchi, Mina" sort="Nakauchi, Mina" uniqKey="Nakauchi M" first="Mina" last="Nakauchi">Mina Nakauchi</name></author><author><name sortKey="Ishizuka, Mariko" sort="Ishizuka, Mariko" uniqKey="Ishizuka M" first="Mariko" last="Ishizuka">Mariko Ishizuka</name></author><author><name sortKey="Hashiguchi, Kazuaki" sort="Hashiguchi, Kazuaki" uniqKey="Hashiguchi K" first="Kazuaki" last="Hashiguchi">Kazuaki Hashiguchi</name></author><author><name sortKey="Hashimoto, Shingo" sort="Hashimoto, Shingo" uniqKey="Hashimoto S" first="Shingo" last="Hashimoto">Shingo Hashimoto</name></author><author><name sortKey="Yoshii, Kentaro" sort="Yoshii, Kentaro" uniqKey="Yoshii K" first="Kentaro" last="Yoshii">Kentaro Yoshii</name></author><author><name sortKey="Asano, Atsushi" sort="Asano, Atsushi" uniqKey="Asano A" first="Atsushi" last="Asano">Atsushi Asano</name></author><author><name sortKey="Agui, Takashi" sort="Agui, Takashi" uniqKey="Agui T" first="Takashi" last="Agui">Takashi Agui</name></author><author><name sortKey="Kogaki, Hiroyuki" sort="Kogaki, Hiroyuki" uniqKey="Kogaki H" first="Hiroyuki" last="Kogaki">Hiroyuki Kogaki</name></author><author><name sortKey="Kurano, Yoshihiro" sort="Kurano, Yoshihiro" uniqKey="Kurano Y" first="Yoshihiro" last="Kurano">Yoshihiro Kurano</name></author><author><name sortKey="Uchida, Yoshiaki" sort="Uchida, Yoshiaki" uniqKey="Uchida Y" first="Yoshiaki" last="Uchida">Yoshiaki Uchida</name></author><author><name sortKey="Fujii, Nobuyuki" sort="Fujii, Nobuyuki" uniqKey="Fujii N" first="Nobuyuki" last="Fujii">Nobuyuki Fujii</name></author><author><name sortKey="Okada, Masahisa" sort="Okada, Masahisa" uniqKey="Okada M" first="Masahisa" last="Okada">Masahisa Okada</name></author><author><name sortKey="Takashima, Ikuo" sort="Takashima, Ikuo" uniqKey="Takashima I" first="Ikuo" last="Takashima">Ikuo Takashima</name></author></analytic><series><title level="j">The Japanese journal of veterinary research</title><idno type="ISSN">0047-1917</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Sequence</term><term>Amino Acids (chemistry)</term><term>Animals</term><term>Antibodies, Monoclonal (chemistry)</term><term>Antibodies, Monoclonal (immunology)</term><term>Antibodies, Viral (chemistry)</term><term>Antibody Specificity</term><term>Blotting, Western (veterinary)</term><term>Chlorocebus aethiops</term><term>Cloning, Molecular</term><term>Coronavirus 229E, Human</term><term>Cross Reactions</term><term>Epitope Mapping</term><term>Fluorescent Antibody Technique, Indirect (veterinary)</term><term>Humans</term><term>Mice</term><term>Nucleocapsid Proteins (chemistry)</term><term>Nucleocapsid Proteins (genetics)</term><term>Nucleocapsid Proteins (immunology)</term><term>Rabbits</term><term>Reverse Transcriptase Polymerase Chain Reaction (veterinary)</term><term>SARS Virus (immunology)</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (veterinary)</term><term>Vero Cells</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Acides aminés ()</term><term>Animaux</term><term>Anticorps antiviraux ()</term><term>Anticorps monoclonaux ()</term><term>Anticorps monoclonaux (immunologie)</term><term>Cartographie épitopique</term><term>Cellules Vero</term><term>Clonage moléculaire</term><term>Coronavirus humain 229E</term><term>Humains</term><term>Lapins</term><term>Protéines nucléocapside ()</term><term>Protéines nucléocapside (génétique)</term><term>Protéines nucléocapside (immunologie)</term><term>RT-PCR (médecine vétérinaire)</term><term>Réactions croisées</term><term>Souris</term><term>Spécificité des anticorps</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (médecine vétérinaire)</term><term>Séquence d'acides aminés</term><term>Technique d'immunofluorescence indirecte (médecine vétérinaire)</term><term>Technique de Western (médecine vétérinaire)</term><term>Virus du SRAS (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Amino Acids</term><term>Antibodies, Monoclonal</term><term>Antibodies, Viral</term><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Monoclonal</term><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines nucléocapside</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps monoclonaux</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="médecine vétérinaire" xml:lang="fr"><term>RT-PCR</term><term>Syndrome respiratoire aigu sévère</term><term>Technique d'immunofluorescence indirecte</term><term>Technique de Western</term></keywords><keywords scheme="MESH" qualifier="veterinary" xml:lang="en"><term>Blotting, Western</term><term>Fluorescent Antibody Technique, Indirect</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Sequence</term><term>Animals</term><term>Antibody Specificity</term><term>Chlorocebus aethiops</term><term>Cloning, Molecular</term><term>Coronavirus 229E, Human</term><term>Cross Reactions</term><term>Epitope Mapping</term><term>Humans</term><term>Mice</term><term>Rabbits</term><term>Vero Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Acides aminés</term><term>Animaux</term><term>Anticorps antiviraux</term><term>Anticorps monoclonaux</term><term>Cartographie épitopique</term><term>Cellules Vero</term><term>Clonage moléculaire</term><term>Coronavirus humain 229E</term><term>Humains</term><term>Lapins</term><term>Protéines nucléocapside</term><term>Réactions croisées</term><term>Souris</term><term>Spécificité des anticorps</term><term>Séquence d'acides aminés</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The sudden emergence of severe acute respiratory syndrome (SARS) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. As no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a SARS epidemic is the rapid diagnosis and isolation of SARS patients. To establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (NP) of SARS-coronavirus (SARS-CoV). On immunofluorescent antibody assay and Western blotting analysis, none of the monoclonal antibodies showed cross-reactivity to authentic and recombinant NPs of human coronavirus (HCoV) 229E strain. To determine the region on the NP molecule where the monoclonal antibodies bind, we generated four truncated recombinant NPs and analyzed the reactivity between monoclonal antibodies and truncated NPs. Two monoclonal antibodies reacted with a truncated NP covering from amino acid residues 111 to 230, and seven reacted with another truncated NP covering from amino acid residues 221 to 340. Epitope mapping analysis indicated that monoclonal antibody SN5-25 recognized the amino acid sequence Q(245)TVTKK(250) On SARS-NP. Within the epitope, Q245, T246, V247, K249, and K250 appeared to form an essential motif for monoclonal antibody SN5-25 to bind. The information about binding sites and epitopes of monoclonal antibodies may be useful for the development of new diagnostic methods for SARS and for analyzing the function of N protein of SARS-CoV.</div></front></TEI><affiliations><list><country><li>Japon</li></country></list><tree><noCountry><name sortKey="Agui, Takashi" sort="Agui, Takashi" uniqKey="Agui T" first="Takashi" last="Agui">Takashi Agui</name><name sortKey="Asano, Atsushi" sort="Asano, Atsushi" uniqKey="Asano A" first="Atsushi" last="Asano">Atsushi Asano</name><name sortKey="Fujii, Nobuyuki" sort="Fujii, Nobuyuki" uniqKey="Fujii N" first="Nobuyuki" last="Fujii">Nobuyuki Fujii</name><name sortKey="Hashiguchi, Kazuaki" sort="Hashiguchi, Kazuaki" uniqKey="Hashiguchi K" first="Kazuaki" last="Hashiguchi">Kazuaki Hashiguchi</name><name sortKey="Hashimoto, Shingo" sort="Hashimoto, Shingo" uniqKey="Hashimoto S" first="Shingo" last="Hashimoto">Shingo Hashimoto</name><name sortKey="Ishizuka, Mariko" sort="Ishizuka, Mariko" uniqKey="Ishizuka M" first="Mariko" last="Ishizuka">Mariko Ishizuka</name><name sortKey="Kogaki, Hiroyuki" sort="Kogaki, Hiroyuki" uniqKey="Kogaki H" first="Hiroyuki" last="Kogaki">Hiroyuki Kogaki</name><name sortKey="Kurano, Yoshihiro" sort="Kurano, Yoshihiro" uniqKey="Kurano Y" first="Yoshihiro" last="Kurano">Yoshihiro Kurano</name><name sortKey="Nakauchi, Mina" sort="Nakauchi, Mina" uniqKey="Nakauchi M" first="Mina" last="Nakauchi">Mina Nakauchi</name><name sortKey="Noda, Hiroshi" sort="Noda, Hiroshi" uniqKey="Noda H" first="Hiroshi" last="Noda">Hiroshi Noda</name><name sortKey="Okada, Masahisa" sort="Okada, Masahisa" uniqKey="Okada M" first="Masahisa" last="Okada">Masahisa Okada</name><name sortKey="Takashima, Ikuo" sort="Takashima, Ikuo" uniqKey="Takashima I" first="Ikuo" last="Takashima">Ikuo Takashima</name><name sortKey="Uchida, Yoshiaki" sort="Uchida, Yoshiaki" uniqKey="Uchida Y" first="Yoshiaki" last="Uchida">Yoshiaki Uchida</name><name sortKey="Yoshii, Kentaro" sort="Yoshii, Kentaro" uniqKey="Yoshii K" first="Kentaro" last="Yoshii">Kentaro Yoshii</name></noCountry><country name="Japon"><noRegion><name sortKey="Kariwa, Hiroaki" sort="Kariwa, Hiroaki" uniqKey="Kariwa H" first="Hiroaki" last="Kariwa">Hiroaki Kariwa</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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