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Banana allergy has been associated with the latex-fruit syndrome. Several IgE-binding components, the relevant ones being proteins of 30-37-kDa, have been detected in banana fruit, but none of them have been isolated and characterized yet.To purify and characterize the 30-37 kDa banana allergens.Fifteen patients allergic to banana were selected on the grounds of a latex-allergic population. Prick by prick tests to this fruit were performed. Total and specific IgE to banana were determined. Banana allergens were isolated by affinity chromatography, followed by cation-exchange chromatography. Their characterization includes N-terminal sequencing, enzymatic activity assays, immunodetection with sera from allergic patients and with antichitinase antibodies, and CAP and immunoblot inhibition tests. Skin prick tests with banana extracts and with the purified allergens were also carried out.Two major IgE-binding proteins of 34 and 32 kDa, also recognized by polyclonal antibodies against chestnut chitinases, were immunodetected in crude banana extracts. Purification and characterization of both proteins have allowed their identification as class I chitinases with an hevein-like domain. Each isolated allergen reached inhibition values higher than 90% in CAP inhibition assays, and fully inhibited the IgE-binding by the crude banana extract when tested by an immunoblot inhibition method. The two purified allergens provoked positive skin prick test responses in more than 50% of the banana-allergic patients.Class I chitinases with an hevein-like domain are major allergens in banana fruit. Their presence in other fruits and nuts, such as avocado and chestnut, could explain the cross-sensitization among these foods.