Purification of enolase from Candida albicans KNIH10 isolated in Korea and application of immunological diagnosis

ABSTRACT

We purified enolase from Candida albicans KNIH10 strain which was isolated from a clinical specimen in Korea. The purified enolase was used to detect anti-Candida antibodies in sera of patients with invasive candidiasis. For purification of enolase from the crude extract prepared by French pressure at 20,000 PSI, the fast performance liquid chromatography (FPLC) using DEAE-sepharose column was used. The elutes at 0.3-0.4 M NaCl in FPLC was purified with homogenity in SDS-PAGE and its enzymatic activity was confirmed in sera of invasive candidiasis with candidemia patient by immunoblotting. The purified enolase indicated no siggnal (100% specificity) in 40 normal human sera and 75% (6/8) sensitivity in sera of candidemic patients with suspicious invasive candidiasis by immunoblotting.