Abstract
A component ofFusarium 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 ofFus 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 ofFus 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 * ofF. solani CF.
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Abbreviations
- DEAE:
-
diethyl aminoethyl
- HPLC:
-
high performance liquid chromatography
- SDS:
-
sodium dodecyl sulphate
- PBS-Tween:
-
phosphate buffer saline containing 0.1% tween
- DTT:
-
dithiothreitol
- TFA:
-
triflouroacetate
- FPLC:
-
fast protein liquid chromatography
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Verma, J., Pasha, S. & Gangal, S.V. Purification and characterization ofFus sI3596, a 65 kd allergen ofFusarium solani . Mol Cell Biochem 131, 157–166 (1994). https://doi.org/10.1007/BF00925952
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DOI: https://doi.org/10.1007/BF00925952