方法:食用昆虫蛋白被越来越多地引入作为替代的可持续食物来源,以满足世界对不断增长的人口的需求。原肌球蛋白是主要的昆虫过敏原;然而,其他潜在的过敏原没有很好地表征,并且提取程序对免疫反应性的影响尚不清楚。
结果:使用五种不同的提取缓冲液提取了来自板球(Achetadomesticus)和黑兵蝇(BSF)(Hermetiaillucens)的不同商业食品的蛋白质。通过SDS-PAGE和使用变应原特异性抗体和甲壳动物变应性患者血清的免疫印迹分析蛋白质。通过质谱分析IgE结合带以及所有30种提取物的完整变应原谱。基于尿素的缓冲液在提取昆虫过敏原方面最有效。虾特异性抗体与板球和BSF的原肌球蛋白的交叉反应性表明虾和昆虫之间具有高度的序列和结构相似性。在这两个物种中都发现了其他独特的过敏原,包括血蓝蛋白,卵黄蛋白原,HSP20,载脂蛋白-III,和几丁质结合蛋白。
结论:确定板球和BSF中潜在的变应原性蛋白及其亚型需要使用基于尿素的方法的特定提取方法。虽然原肌球蛋白是最丰富和免疫反应性的过敏原,确定了七种独特的过敏原,强调需要在食品中检测昆虫物种特异性过敏原。
METHODS: Edible insect proteins are increasingly introduced as an alternative sustainable food source to address the world\'s need to feed the growing population. Tropomyosin is the main insect
allergen; however, additional potential allergens are not well characterized and the impact of extraction procedures on immunological reactivity is unknown.
RESULTS: Proteins from different commercial food products derived from cricket (Acheta domesticus) and black soldier fly (BSF) (Hermetia illucens) are extracted using five different extraction buffers. The proteins are analyzed by SDS-PAGE and immunoblotting using
allergen-specific antibodies and crustacean allergic patient sera. IgE binding bands are analyzed by mass spectrometry as well as the complete
allergen profile of all 30 extracts. Urea-based buffers are most efficient in extracting insect allergens. Shrimp-specific antibody cross-reactivity to tropomyosin from cricket and BSF indicates high sequence and structural similarity between shrimp and insects. Additional unique allergens are identified in both species, including hemocyanin, vitellogenin, HSP20, apolipophorin-III, and chitin-binding protein.
CONCLUSIONS: Identifying potential allergenic proteins and their isoforms in cricket and BSF requires specific extraction approaches using urea-based methods. While tropomyosin is the most abundant and immunoreactive
allergen, seven unique allergens are identified, highlighting the need for insect species-specific
allergen detection in food products.