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Structure. 2013 Jan 8;21(1):167-175. doi: 10.1016/j.str.2012.10.005. Epub 2012 Nov 15.

A structure-based strategy for epitope discovery in Burkholderia pseudomallei OppA antigen.

Author information

1
Department of Biosciences, University of Milan, Milan 20133, Italy.
2
Consiglio Nazionale delle Ricerche, Institute for Chemistry of Molecular Recognition, Department of Computational Biology, Milan 20131, Italy.
3
Institute of Biotechnology and Biomedicine, Universitat Autònoma de Barcelona, Bellaterra 08193, Spain.
4
Center for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
5
Institute of Biotechnology and Biomedicine, Universitat Autònoma de Barcelona, Bellaterra 08193, Spain; Catalan Institution for Research and Advanced Studies, Barcelona 08010, Spain.
6
Consiglio Nazionale delle Ricerche, Institute for Chemistry of Molecular Recognition, Department of Computational Biology, Milan 20131, Italy. Electronic address: giorgio.colombo@icrm.cnr.it.
7
Department of Biosciences, University of Milan, Milan 20133, Italy; Consiglio Nazionale delle Ricerche, Institute of Biophysics, University of Milan, Milan 20133, Italy. Electronic address: martino.bolognesi@unimi.it.

Abstract

We present an approach integrating structural and computational biology with immunological tests to identify epitopes in the OppA antigen from the Gram-negative pathogen Burkholderia pseudomallei, the etiological agent of melioidosis. The crystal structure of OppA(Bp), reported here at 2.1 Å resolution, was the basis for a computational analysis that identified three potential epitopes. In parallel, antigen proteolysis and immunocapturing allowed us to identify three additional peptides. All six potential epitopes were synthesized as free peptides and tested for their immunoreactivity against sera from healthy seronegative, healthy seropositive, and recovered melioidosis patients. Three synthetic peptides allowed the different patient groups to be distinguished, underlining the potential of this approach. Extension of the computational analysis, including energy-based decomposition methods, allowed rationalizing results of the predictive analyses and the immunocapture epitope mapping. Our results illustrate a structure-based epitope discovery process, whose application may expand our perspectives in the diagnostic and vaccine design fields.

PMID:
23159127
DOI:
10.1016/j.str.2012.10.005
[Indexed for MEDLINE]
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