Supplementary Material for: Ani s 11-Like Protein Is a Pepsin- and Heat-Resistant Major Allergen of Anisakis spp. and a Valuable Tool for Anisakis Allergy Component-Resolved Diagnosis

Abstract

<br><em><strong>Background:</strong></em><i>Anisakis simplex</i> is a fish parasite responsible for gastrointestinal and allergic symptoms in humans. The Ani s 11-like protein has been proposed as an <i>Anisakis</i> allergen because its primary structure is similar to that of Ani s 11. The aims of this work were to analyse the frequency of detection of the Ani s 11-like protein and assess its diagnostic value. <b><i>Methods:</i></b> rAni s 11-like protein, rAni s 5 and rAni s 4 were expressed in <i>Escherichia coli</i> and rAni s 1 was produced in <i>Pichia pastoris</i>. Recombinant allergen detection patterns in 37 <i>Anisakis</i>-sensitised patients were determined. The stability to pepsin digestion and heat treatment of rAni s 11-like protein was also analysed by IgE immunoblotting. <b><i>Results:</i></b> Ani s 11-like protein is a major allergen detected by 78% of <i>Anisakis</i>-allergic patients, and 13.5% of patients detect only the rAni s 11-like allergen. This allergen is heat stable because it retains its capability of binding IgE after boiling for 30 min and it is resistant to pepsin digestion for 120 min. <b><i>Conclusions:</i></b> These data indicate that the Ani s 11-like protein is a pepsin- and heat-resistant major allergen (Ani s 11.0201) of <i>Anisakis</i> spp. and a valuable tool for <i>Anisakis</i> allergy component-resolved diagnosis