Since 2018, insects have belonged the category of Novel Foods and the presence of allergens represents one of the main hazards connected to their consumption, also due to the potential cross-reactivity with Arthropoda pan-allergens. In the present work, the allergenicity assessment of black soldier fly and lesser mealworm was performed with a shotgun bottom-up proteomic approach combined with in-silico assessment, followed by IgG- and IgE-immunoblotting experiments. The peptides identified, filtered for their abundance and robustness, belonged mainly to muscle proteins, which represented the most abundant protein group. The relevant potential allergens were in-silico identified by sequence similarity to known allergens, and among them tropomyosin resulted the most abundant insect allergen. IgG-immunoblotting analysis with anti-Tropomyosin I antibodies and IgE-immunoblotting assay with serum from patient allergic to crustacean tropomyosin were performed in order to assess the immunoreactivity in both insects. The immunoassays were carried out also on protein hydrolysates extracted by treating insects with Protease from Bacillus licheniformis (1%, 60 °C, pH 7.5). While IgG-immunoblotting demonstrated the loss of immunoreactivity for both hydrolysates, IgE-immunoblotting showed a partial immunoreactivity preservation, also after hydrolysis, in the case of black soldier fly hydrolysate, and a total loss of immunoreactivity for lesser mealworm hydrolysate.

中文翻译：

---

gun弹枪蛋白质组学，计算机内评估和免疫印迹测定法，用于评估小粉虫，黑战士果蝇及其蛋白水解物的致敏性。

自2018年以来，昆虫已属于新型食品类别，并且由于与节肢动物泛过敏原具有潜在的交叉反应性，因此过敏原的存在是与其食用有关的主要危害之一。在目前的工作中，使用a弹枪自下而上的蛋白质组学方法结合计算机内评估，对黑战士苍蝇和小粉虫进行了变应原性评估，随后进行了IgG和IgE免疫印迹实验。鉴定出的肽由于其丰度和耐用性而被过滤，主要属于代表最大蛋白质组的肌肉蛋白。通过与已知变应原的序列相似性在计算机上鉴定了相关的潜在变应原，其中原肌球蛋白导致了最丰富的昆虫变应原。为了评估在两种昆虫中的免疫反应性，使用抗肌钙蛋白I抗体进行了IgG免疫印迹分析，并使用了对甲壳类原肌球蛋白过敏的患者的血清进行了IgE免疫印迹测定。还对用地衣芽孢杆菌的蛋白酶处理昆虫提取的蛋白质水解产物（1％，60°C，pH 7.5）进行了免疫分析。尽管IgG免疫印迹显示了两种水解产物的免疫反应性都丧失了，但IgE免疫印迹显示了黑士兵果蝇水解产物在水解后仍保留了部分免疫反应性，而较少的粉虫水解产物则完全丧失了免疫反应性。还对用地衣芽孢杆菌的蛋白酶处理昆虫提取的蛋白质水解产物（1％，60°C，pH 7.5）进行了免疫分析。尽管IgG免疫印迹显示了两种水解产物的免疫反应性都丧失了，但IgE免疫印迹显示了黑士兵蝇水解产物在水解后仍保留了部分免疫反应性，而较小的粉虫水解产物则完全丧失了免疫反应性。免疫分析还对通过用蛋白酶处理来自地衣芽孢杆菌的昆虫提取的蛋白质水解物（1％，60°C，pH 7.5）进行。尽管IgG免疫印迹显示了两种水解产物的免疫反应性都丧失了，但IgE免疫印迹显示了黑士兵果蝇水解产物在水解后仍保留了部分免疫反应性，而较少的粉虫水解产物则完全丧失了免疫反应性。