Extraction of cashew allergens for sensitization and challenge.
Proteins were extracted from acetone-defatted cashew flour (Stallergenes Greer, Lenoir, NC) by overnight stirring in PBS 1X, at 4°C. The solution was centrifuged for 30 min at 3000 x g to eliminate insoluble components. Supernatant was frozen at -80°C and lyophilized (Lyofal, Salon de Provence, France). Nitrogen inertization was performed. Solutions for sensitization and oral challenge were obtained by adding the required volume of PBS 1X into vials. The final protein concentration was controlled using Bradford.