A rapid method for the extraction of whole cell proteins from Candida species

A rapid method for the extraction of whole cell proteins from yeasts of the genus Candida has been developed. Stationary phase cells (109) were harvested, washed with phosphate-buffered saline and resuspended in 200 μl of sodium dodecyl sulphate (SDS) sample buffer. The cell suspension was boiled for 20 min and 20 μl were applied directly to a 12.5% polyacrylamide gel. Electrophoresis was performed for 18 h at 75 V, after which time, staining with Coomassie Brilliant Blue R 250 revealed the presence of approximately 30 distinct bands, depending upon the species. This method of extracting whole cell proteins is rapid, easy to perform and has been used with a range of Candida species (viz. C. albicans, C. tropicalis, C. pseudotropicalis, C. parapsilosis, C. lusitaniae, C. krusei and C. glabrata) obtained from both laboratory stocks and clinical samples. It enables the differentiation of a large number of Candida species based upon the protein banding pattern(s) obtained after SDS–polyacrylamide gel electrophoretic analysis