Purification and characterization of Fus sI3596, a 65 kd allergen of Fusarium solani

Abstract

A component of Fusarium solani (F. solani), identified as the major allergen, Fus sI3596 was purified to homogeneity from culture filtrate (CF) by means of anion-exchange column chromatography, gel filtration and FPLC. The homogeneity of Fus sI3596 was assessed by IEF, PAGE, SDS-PAGE (non-reducing), immunoblot and HPLC. Fus sI3596 was isolated as a glycoprotein of MW 65 kd and pI 3.6. The IgE ELISA-inhibition assay after periodate treatment of the fraction showed a lower IgE binding capacity suggesting involvement of carbohydrate moiety in IgE binding reactions of the allergen. Peptide fragments of Fus sI3596 obtained after CNBr and trypsin treatment were analysed by immunoblotting for their allergenicity. This study indicated that there could be at least 3 allergenic determinants in the major allergen, Fus sI3596 of F.solani CF.