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 Candid a albicans, an important human opportunistic pathogen. The full-length codi ng sequence of C, albicans enolase gene was subcloned into the prokaryotic expression vector pDS56/RBSII,His(6)/E-under the control of an inducible pr omoter to produce a His(6)-tagged enolase, The recombinant protein was puri fied to homogeneity by one-step nickel-chelate affinity chromatography, It was recognized by a monoclonal antibody specific for C. albicans enolase, a s well as by anti-enolase antibodies present in human sera (IgG). The recom binant protein promptly elicited antibody in mice and detected immune respo nses in normal human subjects that were comparable to those generated by th e 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.