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.