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. | ![]() |