Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from Plasmodium falciparum
Identifieur interne : 001429 ( Pmc/Checkpoint ); précédent : 001428; suivant : 001430Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from Plasmodium falciparum
Auteurs : Michael Alifrangis [Danemark] ; Inge T. Christensen [Danemark] ; Flemming S. J Rgensen [Danemark] ; Anita M. R Nn [Danemark] ; Jimmy E. Weng [Danemark] ; Ming Chen [Danemark] ; Ib C. Bygbjerg [Danemark] ; Worachart Sirawaraporn [Thaïlande] ; Yaseelan Palarasah [Danemark] ; Claus Koch [Danemark]Source :
- Malaria Journal [ 1475-2875 ] ; 2004.
Abstract
The aim of this study was to develop site-specific antibodies as a tool to capture
A homology model of Pf-DHFR domain was employed to define an epitope for the development of site-specific antibodies against Pf-DHFR. The homology model suggested an exposed loop encompassing amino acid residues 64–100. A synthetic peptide of 37-mers whose sequence corresponded to the sequence of amino acid residues 64–100 of Pf-DHFR was synthesized and used to immunize mice for antibodies. Additionally, polyclonal antibodies recognizing a recombinant DHFR enzyme were produced in rabbits.
Serum from mice immunized with the 37-mer showed strong reactivity against both the immunizing peptide, recombinant DHFR and a preparation of crude antigen from
Url:
DOI: 10.1186/1475-2875-3-16
PubMed: 15193156
PubMed Central: 449723
Affiliations:
Links toward previous steps (curation, corpus...)
Links to Exploration step
PMC:449723Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from <italic>Plasmodium falciparum</italic></title><author><name sortKey="Alifrangis, Michael" sort="Alifrangis, Michael" uniqKey="Alifrangis M" first="Michael" last="Alifrangis">Michael Alifrangis</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Christensen, Inge T" sort="Christensen, Inge T" uniqKey="Christensen I" first="Inge T" last="Christensen">Inge T. Christensen</name><affiliation wicri:level="1"><nlm:aff id="I3">Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen</wicri:regionArea><wicri:noRegion>DK-2100 Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I6">Present address: Novo Nordisk A/S, Novo Nordisk Park, Maaloev, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Present address: Novo Nordisk A/S, Novo Nordisk Park, Maaloev</wicri:regionArea><wicri:noRegion>Maaloev</wicri:noRegion></affiliation></author><author><name sortKey="J Rgensen, Flemming S" sort="J Rgensen, Flemming S" uniqKey="J Rgensen F" first="Flemming S" last="J Rgensen">Flemming S. J Rgensen</name><affiliation wicri:level="1"><nlm:aff id="I3">Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen</wicri:regionArea><wicri:noRegion>DK-2100 Copenhagen</wicri:noRegion></affiliation></author><author><name sortKey="R Nn, Anita M" sort="R Nn, Anita M" uniqKey="R Nn A" first="Anita M" last="R Nn">Anita M. R Nn</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Weng, Jimmy E" sort="Weng, Jimmy E" uniqKey="Weng J" first="Jimmy E" last="Weng">Jimmy E. Weng</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Chen, Ming" sort="Chen, Ming" uniqKey="Chen M" first="Ming" last="Chen">Ming Chen</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Bygbjerg, Ib C" sort="Bygbjerg, Ib C" uniqKey="Bygbjerg I" first="Ib C" last="Bygbjerg">Ib C. Bygbjerg</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Sirawaraporn, Worachart" sort="Sirawaraporn, Worachart" uniqKey="Sirawaraporn W" first="Worachart" last="Sirawaraporn">Worachart Sirawaraporn</name><affiliation wicri:level="1"><nlm:aff id="I5">Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand</nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok</wicri:regionArea><wicri:noRegion>Bangkok</wicri:noRegion></affiliation></author><author><name sortKey="Palarasah, Yaseelan" sort="Palarasah, Yaseelan" uniqKey="Palarasah Y" first="Yaseelan" last="Palarasah">Yaseelan Palarasah</name><affiliation wicri:level="3"><nlm:aff id="I4">Immunological Research Laboratory, State Serum Institute, Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Immunological Research Laboratory, State Serum Institute, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Koch, Claus" sort="Koch, Claus" uniqKey="Koch C" first="Claus" last="Koch">Claus Koch</name><affiliation wicri:level="3"><nlm:aff id="I4">Immunological Research Laboratory, State Serum Institute, Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Immunological Research Laboratory, State Serum Institute, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">15193156</idno><idno type="pmc">449723</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC449723</idno><idno type="RBID">PMC:449723</idno><idno type="doi">10.1186/1475-2875-3-16</idno><date when="2004">2004</date><idno type="wicri:Area/Pmc/Corpus">000564</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000564</idno><idno type="wicri:Area/Pmc/Curation">000564</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000564</idno><idno type="wicri:Area/Pmc/Checkpoint">001429</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">001429</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from <italic>Plasmodium falciparum</italic></title><author><name sortKey="Alifrangis, Michael" sort="Alifrangis, Michael" uniqKey="Alifrangis M" first="Michael" last="Alifrangis">Michael Alifrangis</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Christensen, Inge T" sort="Christensen, Inge T" uniqKey="Christensen I" first="Inge T" last="Christensen">Inge T. Christensen</name><affiliation wicri:level="1"><nlm:aff id="I3">Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen</wicri:regionArea><wicri:noRegion>DK-2100 Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I6">Present address: Novo Nordisk A/S, Novo Nordisk Park, Maaloev, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Present address: Novo Nordisk A/S, Novo Nordisk Park, Maaloev</wicri:regionArea><wicri:noRegion>Maaloev</wicri:noRegion></affiliation></author><author><name sortKey="J Rgensen, Flemming S" sort="J Rgensen, Flemming S" uniqKey="J Rgensen F" first="Flemming S" last="J Rgensen">Flemming S. J Rgensen</name><affiliation wicri:level="1"><nlm:aff id="I3">Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen</wicri:regionArea><wicri:noRegion>DK-2100 Copenhagen</wicri:noRegion></affiliation></author><author><name sortKey="R Nn, Anita M" sort="R Nn, Anita M" uniqKey="R Nn A" first="Anita M" last="R Nn">Anita M. R Nn</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Weng, Jimmy E" sort="Weng, Jimmy E" uniqKey="Weng J" first="Jimmy E" last="Weng">Jimmy E. Weng</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Chen, Ming" sort="Chen, Ming" uniqKey="Chen M" first="Ming" last="Chen">Ming Chen</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Bygbjerg, Ib C" sort="Bygbjerg, Ib C" uniqKey="Bygbjerg I" first="Ib C" last="Bygbjerg">Ib C. Bygbjerg</name><affiliation wicri:level="1"><nlm:aff id="I1">Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen</wicri:regionArea><wicri:noRegion>University of Copenhagen</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Department of Infectious Diseases, Copenhagen University Hospital, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Department of Infectious Diseases, Copenhagen University Hospital</wicri:regionArea><wicri:noRegion>Copenhagen University Hospital</wicri:noRegion></affiliation></author><author><name sortKey="Sirawaraporn, Worachart" sort="Sirawaraporn, Worachart" uniqKey="Sirawaraporn W" first="Worachart" last="Sirawaraporn">Worachart Sirawaraporn</name><affiliation wicri:level="1"><nlm:aff id="I5">Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand</nlm:aff><country xml:lang="fr">Thaïlande</country><wicri:regionArea>Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok</wicri:regionArea><wicri:noRegion>Bangkok</wicri:noRegion></affiliation></author><author><name sortKey="Palarasah, Yaseelan" sort="Palarasah, Yaseelan" uniqKey="Palarasah Y" first="Yaseelan" last="Palarasah">Yaseelan Palarasah</name><affiliation wicri:level="3"><nlm:aff id="I4">Immunological Research Laboratory, State Serum Institute, Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Immunological Research Laboratory, State Serum Institute, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author><author><name sortKey="Koch, Claus" sort="Koch, Claus" uniqKey="Koch C" first="Claus" last="Koch">Claus Koch</name><affiliation wicri:level="3"><nlm:aff id="I4">Immunological Research Laboratory, State Serum Institute, Copenhagen, Denmark</nlm:aff><country xml:lang="fr">Danemark</country><wicri:regionArea>Immunological Research Laboratory, State Serum Institute, Copenhagen</wicri:regionArea><placeName><settlement type="city">Copenhague</settlement><region type="région" nuts="2">Hovedstaden</region></placeName></affiliation></author></analytic><series><title level="j">Malaria Journal</title><idno type="eISSN">1475-2875</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>The aim of this study was to develop site-specific antibodies as a tool to capture <italic>Plasmodium falciparum</italic>-dihydrofolate reductase (Pf-DHFR) from blood samples from <italic>P. falciparum </italic>infected individuals in order to detect, in a sandwich ELISA, structural alterations due to point mutations in the gene coding for Pf-DHFR. Furthermore, we wanted to study the potential use of homology models in general and of Pf-DHFR in particular in predicting antigenic malarial surface epitopes.</p></sec><sec sec-type="methods"><title>Methods</title><p>A homology model of Pf-DHFR domain was employed to define an epitope for the development of site-specific antibodies against Pf-DHFR. The homology model suggested an exposed loop encompassing amino acid residues 64–100. A synthetic peptide of 37-mers whose sequence corresponded to the sequence of amino acid residues 64–100 of Pf-DHFR was synthesized and used to immunize mice for antibodies. Additionally, polyclonal antibodies recognizing a recombinant DHFR enzyme were produced in rabbits.</p></sec><sec><title>Results and conclusions</title><p>Serum from mice immunized with the 37-mer showed strong reactivity against both the immunizing peptide, recombinant DHFR and a preparation of crude antigen from <italic>P. falciparum </italic>infected red blood cells. Five monoclonal antibodies were obtained, one of which showed reactivity towards crude antigen prepared from <italic>P. falciparum </italic>infected red cells. Western blot analysis revealed that both the polyclonal and monoclonal antibodies recognized Pf-DHFR. Our study provides insight into the potential use of homology models in general and of Pf-DHFR in particular in predicting antigenic malarial surface epitopes.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct></biblStruct></listBibl></div1></back></TEI><pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Malar J</journal-id><journal-title>Malaria Journal</journal-title><issn pub-type="epub">1475-2875</issn><publisher><publisher-name>BioMed Central</publisher-name><publisher-loc>London</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">15193156</article-id><article-id pub-id-type="pmc">449723</article-id><article-id pub-id-type="publisher-id">1475-2875-3-16</article-id><article-id pub-id-type="doi">10.1186/1475-2875-3-16</article-id><article-categories><subj-group subj-group-type="heading"><subject>Methodology</subject></subj-group></article-categories><title-group><article-title>Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from <italic>Plasmodium falciparum</italic></article-title></title-group><contrib-group><contrib id="A1" corresp="yes" contrib-type="author"><name><surname>Alifrangis</surname><given-names>Michael</given-names></name><xref ref-type="aff" rid="I1">1</xref><xref ref-type="aff" rid="I2">2</xref><email>malif@biobase.dk</email></contrib><contrib id="A2" contrib-type="author"><name><surname>Christensen</surname><given-names>Inge T</given-names></name><xref ref-type="aff" rid="I3">3</xref><xref ref-type="aff" rid="I6">6</xref><email>itc@novonordisk.com</email></contrib><contrib id="A3" contrib-type="author"><name><surname>Jørgensen</surname><given-names>Flemming S</given-names></name><xref ref-type="aff" rid="I3">3</xref><email>fsj@dfh.dk</email></contrib><contrib id="A4" contrib-type="author"><name><surname>Rønn</surname><given-names>Anita M</given-names></name><xref ref-type="aff" rid="I1">1</xref><xref ref-type="aff" rid="I2">2</xref><email>ARN@euro-alarm.dk</email></contrib><contrib id="A5" contrib-type="author"><name><surname>Weng</surname><given-names>Jimmy E</given-names></name><xref ref-type="aff" rid="I1">1</xref><xref ref-type="aff" rid="I2">2</xref><email>j.weng@immi.ku.dk</email></contrib><contrib id="A6" contrib-type="author"><name><surname>Chen</surname><given-names>Ming</given-names></name><xref ref-type="aff" rid="I1">1</xref><xref ref-type="aff" rid="I2">2</xref><email>cmcmp@rh.dk</email></contrib><contrib id="A7" contrib-type="author"><name><surname>Bygbjerg</surname><given-names>Ib C</given-names></name><xref ref-type="aff" rid="I1">1</xref><xref ref-type="aff" rid="I2">2</xref><email>I.Bygbjerg@pubhealth.ku.dk</email></contrib><contrib id="A8" contrib-type="author"><name><surname>Sirawaraporn</surname><given-names>Worachart</given-names></name><xref ref-type="aff" rid="I5">5</xref><email>Scwsr@mahidol.ac.th</email></contrib><contrib id="A9" contrib-type="author"><name><surname>Palarasah</surname><given-names>Yaseelan</given-names></name><xref ref-type="aff" rid="I4">4</xref><email>manicbatsha@yahoo.com</email></contrib><contrib id="A10" contrib-type="author"><name><surname>Koch</surname><given-names>Claus</given-names></name><xref ref-type="aff" rid="I4">4</xref><email>CK@ssi.dk</email></contrib></contrib-group><aff id="I1"><label>1</label>Centre for Medical Parasitology, Institute of Medical Microbiology and Immunology and Institute of Public Health, University of Copenhagen, Denmark</aff><aff id="I2"><label>2</label>Department of Infectious Diseases, Copenhagen University Hospital, Denmark</aff><aff id="I3"><label>3</label>Department of Medicinal Chemistry, The Danish University of Pharmaceutical Sciences, Universitetsparken 2, DK-2100 Copenhagen, Denmark</aff><aff id="I4"><label>4</label>Immunological Research Laboratory, State Serum Institute, Copenhagen, Denmark</aff><aff id="I5"><label>5</label>Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand</aff><aff id="I6"><label>6</label>Present address: Novo Nordisk A/S, Novo Nordisk Park, Maaloev, Denmark</aff><pub-date pub-type="collection"><year>2004</year></pub-date><pub-date pub-type="epub"><day>12</day><month>6</month><year>2004</year></pub-date><volume>3</volume><fpage>16</fpage><lpage>16</lpage><ext-link ext-link-type="uri" xlink:href="http://www.malariajournal.com/content/3/1/16"></ext-link><history><date date-type="received"><day>27</day><month>1</month><year>2004</year></date><date date-type="accepted"><day>12</day><month>6</month><year>2004</year></date></history><permissions><copyright-statement>Copyright © 2004 Alifrangis et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</copyright-statement><copyright-year>2004</copyright-year><copyright-holder>Alifrangis et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.</copyright-holder></permissions><abstract><sec><title>Background</title><p>The aim of this study was to develop site-specific antibodies as a tool to capture <italic>Plasmodium falciparum</italic>-dihydrofolate reductase (Pf-DHFR) from blood samples from <italic>P. falciparum </italic>infected individuals in order to detect, in a sandwich ELISA, structural alterations due to point mutations in the gene coding for Pf-DHFR. Furthermore, we wanted to study the potential use of homology models in general and of Pf-DHFR in particular in predicting antigenic malarial surface epitopes.</p></sec><sec sec-type="methods"><title>Methods</title><p>A homology model of Pf-DHFR domain was employed to define an epitope for the development of site-specific antibodies against Pf-DHFR. The homology model suggested an exposed loop encompassing amino acid residues 64–100. A synthetic peptide of 37-mers whose sequence corresponded to the sequence of amino acid residues 64–100 of Pf-DHFR was synthesized and used to immunize mice for antibodies. Additionally, polyclonal antibodies recognizing a recombinant DHFR enzyme were produced in rabbits.</p></sec><sec><title>Results and conclusions</title><p>Serum from mice immunized with the 37-mer showed strong reactivity against both the immunizing peptide, recombinant DHFR and a preparation of crude antigen from <italic>P. falciparum </italic>infected red blood cells. Five monoclonal antibodies were obtained, one of which showed reactivity towards crude antigen prepared from <italic>P. falciparum </italic>infected red cells. Western blot analysis revealed that both the polyclonal and monoclonal antibodies recognized Pf-DHFR. Our study provides insight into the potential use of homology models in general and of Pf-DHFR in particular in predicting antigenic malarial surface epitopes.</p></sec></abstract></article-meta></front></pmc><affiliations><list><country><li>Danemark</li><li>Thaïlande</li></country><region><li>Hovedstaden</li></region><settlement><li>Copenhague</li></settlement></list><tree><country name="Danemark"><noRegion><name sortKey="Alifrangis, Michael" sort="Alifrangis, Michael" uniqKey="Alifrangis M" first="Michael" last="Alifrangis">Michael Alifrangis</name></noRegion><name sortKey="Alifrangis, Michael" sort="Alifrangis, Michael" uniqKey="Alifrangis M" first="Michael" last="Alifrangis">Michael Alifrangis</name><name sortKey="Bygbjerg, Ib C" sort="Bygbjerg, Ib C" uniqKey="Bygbjerg I" first="Ib C" last="Bygbjerg">Ib C. Bygbjerg</name><name sortKey="Bygbjerg, Ib C" sort="Bygbjerg, Ib C" uniqKey="Bygbjerg I" first="Ib C" last="Bygbjerg">Ib C. Bygbjerg</name><name sortKey="Chen, Ming" sort="Chen, Ming" uniqKey="Chen M" first="Ming" last="Chen">Ming Chen</name><name sortKey="Chen, Ming" sort="Chen, Ming" uniqKey="Chen M" first="Ming" last="Chen">Ming Chen</name><name sortKey="Christensen, Inge T" sort="Christensen, Inge T" uniqKey="Christensen I" first="Inge T" last="Christensen">Inge T. Christensen</name><name sortKey="Christensen, Inge T" sort="Christensen, Inge T" uniqKey="Christensen I" first="Inge T" last="Christensen">Inge T. Christensen</name><name sortKey="J Rgensen, Flemming S" sort="J Rgensen, Flemming S" uniqKey="J Rgensen F" first="Flemming S" last="J Rgensen">Flemming S. J Rgensen</name><name sortKey="Koch, Claus" sort="Koch, Claus" uniqKey="Koch C" first="Claus" last="Koch">Claus Koch</name><name sortKey="Palarasah, Yaseelan" sort="Palarasah, Yaseelan" uniqKey="Palarasah Y" first="Yaseelan" last="Palarasah">Yaseelan Palarasah</name><name sortKey="R Nn, Anita M" sort="R Nn, Anita M" uniqKey="R Nn A" first="Anita M" last="R Nn">Anita M. R Nn</name><name sortKey="R Nn, Anita M" sort="R Nn, Anita M" uniqKey="R Nn A" first="Anita M" last="R Nn">Anita M. R Nn</name><name sortKey="Weng, Jimmy E" sort="Weng, Jimmy E" uniqKey="Weng J" first="Jimmy E" last="Weng">Jimmy E. Weng</name><name sortKey="Weng, Jimmy E" sort="Weng, Jimmy E" uniqKey="Weng J" first="Jimmy E" last="Weng">Jimmy E. Weng</name></country><country name="Thaïlande"><noRegion><name sortKey="Sirawaraporn, Worachart" sort="Sirawaraporn, Worachart" uniqKey="Sirawaraporn W" first="Worachart" last="Sirawaraporn">Worachart Sirawaraporn</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Pmc/Checkpoint
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001429 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Pmc/Checkpoint/biblio.hfd -nk 001429 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Sante
|area= MersV1
|flux= Pmc
|étape= Checkpoint
|type= RBID
|clé= PMC:449723
|texte= Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from Plasmodium falciparum
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Pmc/Checkpoint/RBID.i -Sk "pubmed:15193156" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Pmc/Checkpoint/biblio.hfd \
| NlmPubMed2Wicri -a MersV1
| This area was generated with Dilib version V0.6.33. |  |