Isolation and characterization of a major allergen in kiwi fruit

Abstract

The isolation of an important allergen in kiwi fruit (Actinidia chinensis) by ion-exchange chromatography (IEC) and micropreparative SDS-PAGE followed by electroelution is reported. The purity of the allergen was analysed by SDS-PAGE and immunoblotting with sera from patients who have an allergy to kiwi. The allergen was shown to have a molecular weight of 43 kDa by SDS-PAGE/immunoblotting and an isoelectric point of approximately 6.9 as estimated by IEC. In accordance with World Health Organization nomenclature, this allergen is called Act c 2. By immunoblot inhibition it was shown that epitopes from different allergens in kiwi fruit are also located on Act c 2. N-terminal amino acid sequencing of 17 amino acid residues did not reveal homology with the major allergens in birch pollen (Bet v 1), apple (Mal d 1) or with other proteins of allergenic plant foods. In addition, the isoelectric point of a 67-kDa allergen in kiwi fruit was estimated to be 7.4 by IEC, but micropreparative isolation of this allergen failed because of its very low content in the fruit.