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Please use this identifier to cite or link to this item: http://repository.li.mahidol.ac.th/dspace/handle/123456789/21374
Title: Homology building as a means to define antigenic epitopes on dihydrofolate reductase (DHFR) from Plasmodium falciparum
Authors: Michael Alifrangis
Inge T. Christensen
Flemming S. Jørgensen
Anita M. Rønn
Jimmy E. Weng
Ming Chen
Ib C. Bygbjerg
Worachart Sirawaraporn
Yaseelan Palarasah
Claus Koch
Kobenhavns Universitet
Copenhagen University Hospital
University of Copenhagen, Faculty of Pharmaceutical Sciences
Statens Serum Institut
Mahidol University
Novo Nordisk AS
Keywords: Immunology and Microbiology;Medicine
Issue Date: 12-Jun-2004
Citation: Malaria Journal. Vol.3, (2004)
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.
URI: https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=4844228783&origin=inward
http://repository.li.mahidol.ac.th/dspace/handle/123456789/21374
ISSN: 14752875
Appears in Collections:Scopus 2001-2005

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