Protein extraction strategies shape shellfish allergen landscapes: Implications for seafood safety, allergy diagnosis and risk management

Le résumé fourni par la source

Shellfish is a leading cause of life-threatening food anaphylaxis. Protein extraction is a critical step in allergenic protein (allergen) detection and quantification. This study evaluates the impact of protein extraction strategies on allergen detection in the two most widely consumed shellfish species, black tiger prawn (Penaeus monodon) and white leg prawn (Litopenaeus vannamei). Proteins were extracted from raw and boiled muscle tissues in four different buffers: urea-based, sodium dodecyl sulphate (SDS)-based, SDS-based with reducing agent, and phosphate-buffered saline (PBS), the most commonly used buffer in allergy investigations. Protein abundances were analysed using SDS-PAGE and label-free liquid chromatography-tandem mass spectrometry. Allergenicity was predicted using AllerCatPro and subsequently evaluated for antibody-based immunoreactivity. Protein extraction strategies significantly influenced proteome coverages, IgE-antibody binding, allergen diversities and relative abundances. Although PBS-based extracts yielded the highest overall relative allergen abundances (~84-95%), they generated the lowest allergen diversities and reduced relative abundances of key allergens, including tropomyosin and myosin light chain from raw tissues and arginine kinase from boiled tissues. In contrast, denaturing extracts enabled broader allergen representation, with urea-based extracts from boiled tissues generating the most comprehensive allergen repertoire. In addition to well-characterised allergens, up to 72 low-abundant proteins with strong bioinformatic evidence of allergenicity were identified, with their relative recovery varying across extraction strategies. Optimised protein extraction improves allergen detection workflows, supporting the discovery of underreported and low-abundant candidate allergens for future immunological validation, and facilitating more comprehensive risk assessment strategies.

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