MersV1, Main, Merge, bibRecord, 003091

Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from Plasmodium falciparum

Identifieur interne : 003091 ( Main/Merge ); précédent : 003090; suivant : 003092

Homology 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.

RBID : PMC:449723

Descripteurs français

KwdFr :
- Acides aminés (), Animaux, Anticorps antiprotozoaires (immunologie), Anticorps antiprotozoaires (sang), Anticorps monoclonaux (immunologie), Antigènes de protozoaire (), Antigènes de protozoaire (immunologie), Cartographie épitopique, Dihydrofolate reductase (), Dihydrofolate reductase (immunologie), Humains, Lapins, Modèles moléculaires, Plasmodium falciparum (enzymologie), Plasmodium falciparum (immunologie), Souris, Sérums immuns (immunologie), Technique de Western, Test ELISA, Électrophorèse sur gel de polyacrylamide, Épitopes (), Épitopes (analyse), Épitopes (immunologie).
MESH :
- analyse : Épitopes.
- enzymologie : Plasmodium falciparum.
- immunologie : Anticorps antiprotozoaires, Anticorps monoclonaux, Antigènes de protozoaire, Dihydrofolate reductase, Plasmodium falciparum, Sérums immuns, Épitopes.
- sang : Anticorps antiprotozoaires.
- Acides aminés, Animaux, Antigènes de protozoaire, Cartographie épitopique, Dihydrofolate reductase, Humains, Lapins, Modèles moléculaires, Souris, Technique de Western, Test ELISA, Électrophorèse sur gel de polyacrylamide, Épitopes.

English descriptors

KwdEn :
- Amino Acids (chemistry), Animals, Antibodies, Monoclonal (immunology), Antibodies, Protozoan (blood), Antibodies, Protozoan (immunology), Antigens, Protozoan (chemistry), Antigens, Protozoan (immunology), Blotting, Western, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Epitope Mapping, Epitopes (analysis), Epitopes (chemistry), Epitopes (immunology), Humans, Immune Sera (immunology), Mice, Models, Molecular, Plasmodium falciparum (enzymology), Plasmodium falciparum (immunology), Rabbits, Tetrahydrofolate Dehydrogenase (chemistry), Tetrahydrofolate Dehydrogenase (immunology).
MESH :
- chemical , analysis : Epitopes.
- chemical , blood : Antibodies, Protozoan.
- chemical , chemistry : Amino Acids, Antigens, Protozoan, Epitopes, Tetrahydrofolate Dehydrogenase.
- chemical , immunology : Antibodies, Monoclonal, Antibodies, Protozoan, Antigens, Protozoan, Epitopes, Immune Sera, Tetrahydrofolate Dehydrogenase.
- enzymology : Plasmodium falciparum.
- immunology : Plasmodium falciparum.
- Animals, Blotting, Western, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Epitope Mapping, Humans, Mice, Models, Molecular, Rabbits.

Abstract

Background

The aim of this study was to develop site-specific antibodies as a tool to capture Plasmodium falciparum-dihydrofolate reductase (Pf-DHFR) from blood samples from P. falciparum 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.

Methods

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.

Results and conclusions

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 P. falciparum infected red blood cells. Five monoclonal antibodies were obtained, one of which showed reactivity towards crude antigen prepared from P. falciparum 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.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC449723

DOI: 10.1186/1475-2875-3-16
PubMed: 15193156
PubMed Central: 449723

Links to Exploration step

PMC:449723

Le document en format XML

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<author><name sortKey="Weng, Jimmy E" sort="Weng, Jimmy E" uniqKey="Weng J" first="Jimmy E" last="Weng">Jimmy E. Weng</name>
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<author><name sortKey="Chen, Ming" sort="Chen, Ming" uniqKey="Chen M" first="Ming" last="Chen">Ming Chen</name>
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<author><name sortKey="Bygbjerg, Ib C" sort="Bygbjerg, Ib C" uniqKey="Bygbjerg I" first="Ib C" last="Bygbjerg">Ib C. Bygbjerg</name>
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<author><name sortKey="Sirawaraporn, Worachart" sort="Sirawaraporn, Worachart" uniqKey="Sirawaraporn W" first="Worachart" last="Sirawaraporn">Worachart Sirawaraporn</name>
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<country xml:lang="fr">Thaïlande</country>
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<author><name sortKey="Palarasah, Yaseelan" sort="Palarasah, Yaseelan" uniqKey="Palarasah Y" first="Yaseelan" last="Palarasah">Yaseelan Palarasah</name>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acids (chemistry)</term>
<term>Animals</term>
<term>Antibodies, Monoclonal (immunology)</term>
<term>Antibodies, Protozoan (blood)</term>
<term>Antibodies, Protozoan (immunology)</term>
<term>Antigens, Protozoan (chemistry)</term>
<term>Antigens, Protozoan (immunology)</term>
<term>Blotting, Western</term>
<term>Electrophoresis, Polyacrylamide Gel</term>
<term>Enzyme-Linked Immunosorbent Assay</term>
<term>Epitope Mapping</term>
<term>Epitopes (analysis)</term>
<term>Epitopes (chemistry)</term>
<term>Epitopes (immunology)</term>
<term>Humans</term>
<term>Immune Sera (immunology)</term>
<term>Mice</term>
<term>Models, Molecular</term>
<term>Plasmodium falciparum (enzymology)</term>
<term>Plasmodium falciparum (immunology)</term>
<term>Rabbits</term>
<term>Tetrahydrofolate Dehydrogenase (chemistry)</term>
<term>Tetrahydrofolate Dehydrogenase (immunology)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Acides aminés ()</term>
<term>Animaux</term>
<term>Anticorps antiprotozoaires (immunologie)</term>
<term>Anticorps antiprotozoaires (sang)</term>
<term>Anticorps monoclonaux (immunologie)</term>
<term>Antigènes de protozoaire ()</term>
<term>Antigènes de protozoaire (immunologie)</term>
<term>Cartographie épitopique</term>
<term>Dihydrofolate reductase ()</term>
<term>Dihydrofolate reductase (immunologie)</term>
<term>Humains</term>
<term>Lapins</term>
<term>Modèles moléculaires</term>
<term>Plasmodium falciparum (enzymologie)</term>
<term>Plasmodium falciparum (immunologie)</term>
<term>Souris</term>
<term>Sérums immuns (immunologie)</term>
<term>Technique de Western</term>
<term>Test ELISA</term>
<term>Électrophorèse sur gel de polyacrylamide</term>
<term>Épitopes ()</term>
<term>Épitopes (analyse)</term>
<term>Épitopes (immunologie)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Epitopes</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antibodies, Protozoan</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Amino Acids</term>
<term>Antigens, Protozoan</term>
<term>Epitopes</term>
<term>Tetrahydrofolate Dehydrogenase</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Monoclonal</term>
<term>Antibodies, Protozoan</term>
<term>Antigens, Protozoan</term>
<term>Epitopes</term>
<term>Immune Sera</term>
<term>Tetrahydrofolate Dehydrogenase</term>
</keywords>
<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Épitopes</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Plasmodium falciparum</term>
</keywords>
<keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Plasmodium falciparum</term>
</keywords>
<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiprotozoaires</term>
<term>Anticorps monoclonaux</term>
<term>Antigènes de protozoaire</term>
<term>Dihydrofolate reductase</term>
<term>Plasmodium falciparum</term>
<term>Sérums immuns</term>
<term>Épitopes</term>
</keywords>
<keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Plasmodium falciparum</term>
</keywords>
<keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>Anticorps antiprotozoaires</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Blotting, Western</term>
<term>Electrophoresis, Polyacrylamide Gel</term>
<term>Enzyme-Linked Immunosorbent Assay</term>
<term>Epitope Mapping</term>
<term>Humans</term>
<term>Mice</term>
<term>Models, Molecular</term>
<term>Rabbits</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Acides aminés</term>
<term>Animaux</term>
<term>Antigènes de protozoaire</term>
<term>Cartographie épitopique</term>
<term>Dihydrofolate reductase</term>
<term>Humains</term>
<term>Lapins</term>
<term>Modèles moléculaires</term>
<term>Souris</term>
<term>Technique de Western</term>
<term>Test ELISA</term>
<term>Électrophorèse sur gel de polyacrylamide</term>
<term>Épitopes</term>
</keywords>
</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>
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</listBibl>
</div1>
</back>
</TEI>
</record>

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Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from Plasmodium falciparum

Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from Plasmodium falciparum

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