PURIFICATION AND CHARACTERIZATION OF CELL WALLMANNOPRO TEINS OF CANDIDA ALB/CANS USING INTACT CELL METHOD

Virulence of the opportunistic yeast, Candida albieans, involves the interplay of many complex changes including the yeast-hyphae transition, which mainly involves protein changes. Cell wall mannoproteins are found to be the main cause of adherence of C. albieans to epithel ial cells in the first step of an infection process. In the present study, cell wall mannoproteins of intact yeast were purified using a simple treatment of yeast with mercaptoethanol and sodium dodecyl sulfate followed by Concanavalin A chromatography. Both electrophoretic analysis of the column effluent and Western blot analysis using polyclonal and monoclonal antibodies showed the presence of mannoproteins with molecular weight in the range of30-50 kDa. Dot blot analysis of the purified antigen with the polyclonal and monoclonal antibodies prepared in this study showed that outer membrane mannoprotein antigens were obtained successfully following the above simple purification strategy