[Development of monoclonal antibodies against SARS-CoV and identification of antigenic epitopes].
Identifieur interne : 002647 ( PubMed/Corpus ); précédent : 002646; suivant : 002648[Development of monoclonal antibodies against SARS-CoV and identification of antigenic epitopes].
Auteurs : Yan-Jun Zhou ; Rong-Hong Hua ; Yun-Feng Wang ; Tong-Qing An ; Jin-Xia Liu ; Jin-Yu Yang ; Yu-Zhuo Hua ; Guang-Zhi TongSource :
- Sheng wu gong cheng xue bao = Chinese journal of biotechnology [ 1000-3061 ] ; 2005.
English descriptors
- KwdEn :
- Animals, Antibodies, Monoclonal (biosynthesis), Antibodies, Monoclonal (immunology), Antibodies, Viral (biosynthesis), Antibodies, Viral (immunology), Antibody Specificity, Epitopes (genetics), Escherichia coli (genetics), Escherichia coli (metabolism), Female, Humans, Hybridomas (metabolism), Membrane Glycoproteins (immunology), Mice, Mice, Inbred BALB C, Recombinant Fusion Proteins (biosynthesis), Recombinant Fusion Proteins (genetics), Recombinant Fusion Proteins (immunology), SARS Virus (immunology), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (immunology).
- MESH :
- chemical , biosynthesis : Antibodies, Monoclonal, Antibodies, Viral, Recombinant Fusion Proteins.
- chemical , genetics : Epitopes, Recombinant Fusion Proteins.
- chemical , immunology : Antibodies, Monoclonal, Antibodies, Viral, Membrane Glycoproteins, Recombinant Fusion Proteins, Viral Envelope Proteins.
- genetics : Escherichia coli.
- immunology : SARS Virus.
- metabolism : Escherichia coli, Hybridomas.
- Animals, Antibody Specificity, Female, Humans, Mice, Mice, Inbred BALB C, Spike Glycoprotein, Coronavirus.
Abstract
Based on the genomic sequence of SARS-CoV strain BJ101, antigenic immunodominant genes coding for the structure proteins of SARS-CoV were predicted by bio-informatics methods, and two chimeric genes A and B with multi-immunodominants lined up by Gly-Pro-Gly linker were synthesized. The chimeric genes were cloned into plasmid pGEX-6p-1 and expressed in E. coli with IPGT inducing. BALB/c mice were immunized with the purified recombinant fusion protein. The specificity of monoclonal antibodies were tested with a commercial ELISA kit for detecting antibody against SARS-CoV. The results showed that two peptides with molecular weights of 34kD and 35kD expressed by the two chimeric genes could be recognized by SARS patient convalescent serum in Western blot. Six positive hybridoma cell lines stably secreting monoclonal antibodies were selected. The subtype of monoclonal antibody D3C5 is IgG2a, and subtypes of all other five monoclonal antibodies are IgG1. Light chains of all monoclonal antibodies are kappa. With a commercial SARS-CoV antibodies detection ELISA kit, five out of six monoclonal antibodies were positively recognized. In western blot analysis with inactived virus cultures, D3D1 specifically recognized a band of about 180 kD. To further analyse the epitopes corresponding to the monoclonal antibodies, six oligoes (S1-S6) from S gene were synthesized and expressed. The results showed that the monoclonal antibodies D3D1 and D3C5 specifically recognized expression product of S2 and S5 oligoes, respectively. The S2 and S5 oligoes are corresponding to 447-458aa and 789-799aa of SARS-CoV S protein respectively.
PubMed: 16013477
Links to Exploration step
pubmed:16013477Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">[Development of monoclonal antibodies against SARS-CoV and identification of antigenic epitopes].</title><author><name sortKey="Zhou, Yan Jun" sort="Zhou, Yan Jun" uniqKey="Zhou Y" first="Yan-Jun" last="Zhou">Yan-Jun Zhou</name><affiliation><nlm:affiliation>National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Science, Harbin 150001, China.</nlm:affiliation></affiliation></author><author><name sortKey="Hua, Rong Hong" sort="Hua, Rong Hong" uniqKey="Hua R" first="Rong-Hong" last="Hua">Rong-Hong Hua</name></author><author><name sortKey="Wang, Yun Feng" sort="Wang, Yun Feng" uniqKey="Wang Y" first="Yun-Feng" last="Wang">Yun-Feng Wang</name></author><author><name sortKey="An, Tong Qing" sort="An, Tong Qing" uniqKey="An T" first="Tong-Qing" last="An">Tong-Qing An</name></author><author><name sortKey="Liu, Jin Xia" sort="Liu, Jin Xia" uniqKey="Liu J" first="Jin-Xia" last="Liu">Jin-Xia Liu</name></author><author><name sortKey="Yang, Jin Yu" sort="Yang, Jin Yu" uniqKey="Yang J" first="Jin-Yu" last="Yang">Jin-Yu Yang</name></author><author><name sortKey="Hua, Yu Zhuo" sort="Hua, Yu Zhuo" uniqKey="Hua Y" first="Yu-Zhuo" last="Hua">Yu-Zhuo Hua</name></author><author><name sortKey="Tong, Guang Zhi" sort="Tong, Guang Zhi" uniqKey="Tong G" first="Guang-Zhi" last="Tong">Guang-Zhi Tong</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:16013477</idno><idno type="pmid">16013477</idno><idno type="wicri:Area/PubMed/Corpus">002647</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002647</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">[Development of monoclonal antibodies against SARS-CoV and identification of antigenic epitopes].</title><author><name sortKey="Zhou, Yan Jun" sort="Zhou, Yan Jun" uniqKey="Zhou Y" first="Yan-Jun" last="Zhou">Yan-Jun Zhou</name><affiliation><nlm:affiliation>National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Science, Harbin 150001, China.</nlm:affiliation></affiliation></author><author><name sortKey="Hua, Rong Hong" sort="Hua, Rong Hong" uniqKey="Hua R" first="Rong-Hong" last="Hua">Rong-Hong Hua</name></author><author><name sortKey="Wang, Yun Feng" sort="Wang, Yun Feng" uniqKey="Wang Y" first="Yun-Feng" last="Wang">Yun-Feng Wang</name></author><author><name sortKey="An, Tong Qing" sort="An, Tong Qing" uniqKey="An T" first="Tong-Qing" last="An">Tong-Qing An</name></author><author><name sortKey="Liu, Jin Xia" sort="Liu, Jin Xia" uniqKey="Liu J" first="Jin-Xia" last="Liu">Jin-Xia Liu</name></author><author><name sortKey="Yang, Jin Yu" sort="Yang, Jin Yu" uniqKey="Yang J" first="Jin-Yu" last="Yang">Jin-Yu Yang</name></author><author><name sortKey="Hua, Yu Zhuo" sort="Hua, Yu Zhuo" uniqKey="Hua Y" first="Yu-Zhuo" last="Hua">Yu-Zhuo Hua</name></author><author><name sortKey="Tong, Guang Zhi" sort="Tong, Guang Zhi" uniqKey="Tong G" first="Guang-Zhi" last="Tong">Guang-Zhi Tong</name></author></analytic><series><title level="j">Sheng wu gong cheng xue bao = Chinese journal of biotechnology</title><idno type="ISSN">1000-3061</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Monoclonal (biosynthesis)</term><term>Antibodies, Monoclonal (immunology)</term><term>Antibodies, Viral (biosynthesis)</term><term>Antibodies, Viral (immunology)</term><term>Antibody Specificity</term><term>Epitopes (genetics)</term><term>Escherichia coli (genetics)</term><term>Escherichia coli (metabolism)</term><term>Female</term><term>Humans</term><term>Hybridomas (metabolism)</term><term>Membrane Glycoproteins (immunology)</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Recombinant Fusion Proteins (biosynthesis)</term><term>Recombinant Fusion Proteins (genetics)</term><term>Recombinant Fusion Proteins (immunology)</term><term>SARS Virus (immunology)</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (immunology)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Antibodies, Monoclonal</term><term>Antibodies, Viral</term><term>Recombinant Fusion Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Epitopes</term><term>Recombinant Fusion Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Monoclonal</term><term>Antibodies, Viral</term><term>Membrane Glycoproteins</term><term>Recombinant Fusion Proteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Escherichia coli</term><term>Hybridomas</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Antibody Specificity</term><term>Female</term><term>Humans</term><term>Mice</term><term>Mice, Inbred BALB C</term><term>Spike Glycoprotein, Coronavirus</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Based on the genomic sequence of SARS-CoV strain BJ101, antigenic immunodominant genes coding for the structure proteins of SARS-CoV were predicted by bio-informatics methods, and two chimeric genes A and B with multi-immunodominants lined up by Gly-Pro-Gly linker were synthesized. The chimeric genes were cloned into plasmid pGEX-6p-1 and expressed in E. coli with IPGT inducing. BALB/c mice were immunized with the purified recombinant fusion protein. The specificity of monoclonal antibodies were tested with a commercial ELISA kit for detecting antibody against SARS-CoV. The results showed that two peptides with molecular weights of 34kD and 35kD expressed by the two chimeric genes could be recognized by SARS patient convalescent serum in Western blot. Six positive hybridoma cell lines stably secreting monoclonal antibodies were selected. The subtype of monoclonal antibody D3C5 is IgG2a, and subtypes of all other five monoclonal antibodies are IgG1. Light chains of all monoclonal antibodies are kappa. With a commercial SARS-CoV antibodies detection ELISA kit, five out of six monoclonal antibodies were positively recognized. In western blot analysis with inactived virus cultures, D3D1 specifically recognized a band of about 180 kD. To further analyse the epitopes corresponding to the monoclonal antibodies, six oligoes (S1-S6) from S gene were synthesized and expressed. The results showed that the monoclonal antibodies D3D1 and D3C5 specifically recognized expression product of S2 and S5 oligoes, respectively. The S2 and S5 oligoes are corresponding to 447-458aa and 789-799aa of SARS-CoV S protein respectively.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16013477</PMID><DateCompleted><Year>2009</Year><Month>07</Month><Day>22</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>15</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">1000-3061</ISSN><JournalIssue CitedMedium="Print"><Volume>21</Volume><Issue>2</Issue><PubDate><Year>2005</Year><Month>Mar</Month></PubDate></JournalIssue><Title>Sheng wu gong cheng xue bao = Chinese journal of biotechnology</Title><ISOAbbreviation>Sheng Wu Gong Cheng Xue Bao</ISOAbbreviation></Journal><ArticleTitle>[Development of monoclonal antibodies against SARS-CoV and identification of antigenic epitopes].</ArticleTitle><Pagination><MedlinePgn>211-5</MedlinePgn></Pagination><Abstract><AbstractText>Based on the genomic sequence of SARS-CoV strain BJ101, antigenic immunodominant genes coding for the structure proteins of SARS-CoV were predicted by bio-informatics methods, and two chimeric genes A and B with multi-immunodominants lined up by Gly-Pro-Gly linker were synthesized. The chimeric genes were cloned into plasmid pGEX-6p-1 and expressed in E. coli with IPGT inducing. BALB/c mice were immunized with the purified recombinant fusion protein. The specificity of monoclonal antibodies were tested with a commercial ELISA kit for detecting antibody against SARS-CoV. The results showed that two peptides with molecular weights of 34kD and 35kD expressed by the two chimeric genes could be recognized by SARS patient convalescent serum in Western blot. Six positive hybridoma cell lines stably secreting monoclonal antibodies were selected. The subtype of monoclonal antibody D3C5 is IgG2a, and subtypes of all other five monoclonal antibodies are IgG1. Light chains of all monoclonal antibodies are kappa. With a commercial SARS-CoV antibodies detection ELISA kit, five out of six monoclonal antibodies were positively recognized. In western blot analysis with inactived virus cultures, D3D1 specifically recognized a band of about 180 kD. To further analyse the epitopes corresponding to the monoclonal antibodies, six oligoes (S1-S6) from S gene were synthesized and expressed. The results showed that the monoclonal antibodies D3D1 and D3C5 specifically recognized expression product of S2 and S5 oligoes, respectively. The S2 and S5 oligoes are corresponding to 447-458aa and 789-799aa of SARS-CoV S protein respectively.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Zhou</LastName><ForeName>Yan-Jun</ForeName><Initials>YJ</Initials><AffiliationInfo><Affiliation>National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Science, Harbin 150001, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hua</LastName><ForeName>Rong-Hong</ForeName><Initials>RH</Initials></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Yun-Feng</ForeName><Initials>YF</Initials></Author><Author ValidYN="Y"><LastName>An</LastName><ForeName>Tong-Qing</ForeName><Initials>TQ</Initials></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Jin-Xia</ForeName><Initials>JX</Initials></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Jin-Yu</ForeName><Initials>JY</Initials></Author><Author ValidYN="Y"><LastName>Hua</LastName><ForeName>Yu-Zhuo</ForeName><Initials>YZ</Initials></Author><Author ValidYN="Y"><LastName>Tong</LastName><ForeName>Guang-Zhi</ForeName><Initials>GZ</Initials></Author></AuthorList><Language>chi</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>China</Country><MedlineTA>Sheng Wu Gong Cheng Xue Bao</MedlineTA><NlmUniqueID>9426463</NlmUniqueID><ISSNLinking>1000-3061</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000911">Antibodies, Monoclonal</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000914">Antibodies, Viral</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000939">Epitopes</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C578553">MHV surface projection glycoprotein</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D008562">Membrane Glycoproteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011993">Recombinant Fusion Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D064370">Spike Glycoprotein, Coronavirus</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D014759">Viral Envelope Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C578557">spike glycoprotein, SARS-CoV</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000818" MajorTopicYN="N">Animals</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000911" MajorTopicYN="N">Antibodies, Monoclonal</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="Y">biosynthesis</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D000914" MajorTopicYN="N">Antibodies, Viral</DescriptorName><QualifierName UI="Q000096" MajorTopicYN="N">biosynthesis</QualifierName><QualifierName UI="Q000276" MajorTopicYN="N">immunology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D000918" MajorTopicYN="N">Antibody Specificity</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000939" MajorTopicYN="Y">Epitopes</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004926" MajorTopicYN="N">Escherichia coli</DescriptorName><QualifierName UI="Q000235" 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PubStatus="medline"><Year>2009</Year><Month>7</Month><Day>23</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2005</Year><Month>7</Month><Day>15</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">16013477</ArticleId></ArticleIdList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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