Generation of a highly immunogenic recombinant enolase of the human opportunistic pathogen Candida albicans.
Enolase, a 46 kDa glycolytic enzyme, is an immunodominant antigen of Candida albicans, an important human opportunistic pathogen. The full-length coding sequence of C. albicans enolase gene was subcloned into the prokaryotic expression vector pDS56/RBSII,His6/E- under the control of an inducible promoter to produce a His6-tagged enolase. The recombinant protein was purified to homogeneity by one-step nickel-chelate affinity chromatography. It was recognized by a monoclonal antibody specific for C. albicans enolase, as well as by anti-enolase antibodies present in human sera (IgG). The recombinant protein promptly elicited antibody in mice and detected immune responses in normal human subjects that were comparable to those generated by the native C. albicans enolase. Thus this new recombinant enolase constitutes a valuable reagent for studying the possible role of this protein in anti-Candida immune response.
Sandini
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S
Laboratorio di Batteriologia e Micologia Medica, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy.
Melchionna
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R
Arancia
S
S
Gomez
M J
MJ
La Valle
R
R
D000818:Animals; D000941:Antigens / Q000276:immunology; D002176:Candida albicans / Q000201:enzymology* / Q000472:pathogenicity; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000276:immunology; D051379:Mice; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
D000941:Antigens; D011994:Recombinant Proteins; D005982:Glutathione Transferase; D010751:Phosphopyruvate Hydratase