Assessment of Allergenic Potential for Cross‐Reactivity of Olive ( Olea europaea ) Pit Xylo‐Oligosaccharide‐Rich Extract With Legislated Allergens i

ABSTRACT

Novel food sources must undergo allergenicity assessments to ensure consumer safety and regulatory compliance. Xylo‐oligosaccharide‐rich extract (XOS) derived from the valorization of olive ( Olea europaea ) pit is categorized as a novel food in the context of European Union (EU) food laws. Despite the functional benefits of xylo‐oligosaccharides such as prebiotic, gut modulation, and weight management being a well‐established area, there is a lack of information on the allergenic potential of olive pit‐derived XOS extract. The study aimed to evaluate the cross‐reactivity of olive pit‐derived XOS extract with other allergens through both in vitro and in silico approaches. In vitro cross‐reactivity was assessed using enzyme‐linked immunosorbent assay (ELISA) against eight EU‐legislated allergens: wheat (gluten), milk, hazelnut, pistachio, soy, almond, lupin, and peanut. In silico allergenicity prediction was performed by analyzing the XOS extract amino acid sequence homology with known allergens. In the in vitro tests, hazelnut exhibited the highest cross‐reactivity (0–2.81 ppm), followed by gluten (0–2.10 ppm), almond (0–1.167 ppm), and pistachio (0.210–0.800 ppm). Milk showed minimal cross‐reactivity (0–0.07 ppm), while soy and lupin responses were below detection limits. The in silico analyses revealed that 81% of the 83 amino acid sequences showed no evidence of allergenicity, 7% showed weak evidence, and 12% indicated strong allergenic potential. The findings of the study suggest that olive pit‐derived XOS extract may contain proteins with the potential for cross‐reactivity with certain allergens, corroborating emerging concerns about fruit‐derived allergenic responses. Further clinical studies are recommended to generate additional insights on this preliminary investigation.