Abstract:Jug r 2 is the major allergen of walnut. And the linear epitopes on Jug r 2 is a key factor in triggering a severe allergic reaction. Hence, the research on the efficient strategy for analyzing the linear epitopes of Jug r 2 is necessary. In our study, two immunoinformatic tools including DNAStar Protean and ABCpred were used to predict the B cells linear epitopes on Jug r 2. In vitro simulated gastrointestinal digestion and HPLC-MS/MS were used to identify anti-digestion peptides. And anti-digestion peptides were located on the 3D model of Jug r 2. The results showed that eight predicted B cells linear epitopes were AA186-199, AA226-230, AA257-263, AA284-289, AA373-382, AA389-398, AA408-414, AA472-487; eleven anti-digestion peptides were AA215~220, AA250~260, AA323~337, AA351~356, AA363~388, AA428~438, AA470~483, AA488~513, AA514~526, AA527~541, AA545~572. Of all anti-digestion peptides, four coincided with the predicted or true B cells linear epitope sequences and eight coincided with true T cells epitopes sequences. Anti-digestion peptides showed good antigenic properties in hydrophilic and flexible on the localization of Jug r 2 3D structure. Therefore, the anti-digestion property of food allergens may be a promising direction for studying linear epitopes of Jug r 2.