![]() ![]() From Figure 1B, we found that between 180 and 195 nm, the changes in the α–helix, β–turn, and random coil were relatively stable. The CD was used to determine the structure of the protein solution, and the CD spectrum in the range of 180–260 nm was provided in the protein secondary structure. The sequences of Ana o 2 and Ana o 3 are as follows: The measured fragment sequences were compared with those obtained from the NCBI database, and two of them were identified to match the sequences of Ana o 2 and Ana o 3. The number of peptides obtained from UniProt- Anacardium occidentale was 147, and from the NCBI- Anacardium occidentale, it was 148. A total of 60,842 profiles and 5 proteins were obtained. The mass spectrometric profile data were compared to the UniProt-Anacardium Occidentale (downloaded on 20 April 2019) and NCBI-Anacardium Occidentale databases. The Ana o 2 and Ana o 3 allergens were also identified. The impact of detection during processing can thus be effectively avoided. The obtained data were applied to the database to obtain the corresponding known protein and to determine the sequence of the protein to be measured, as shown in Figure 1A. Specifically, a protein was cut into small fragments by specific enzymatic or chemical hydrolysis, and the molecular weight of each product was detected using mass spectrometry. The main advantage of mass spectrometry in the detection of food allergens is the sequence determination of the peptides and proteins by the protein spectrum. The key allergenic epitopes were chosen, providing important information for the study of cashew nut allergens. Furthermore, the phage display method was extremely effective in identifying the allergenic epitopes of cashew nut proteins. Conclusions: The bioinformatic epitope prediction produced subpar results in this study. Discussion: The Ana o 3 epitopes accounted for more than 40% of the total amino acid sequence of the protein thus, Ana o 3 is potentially more allergenic than Ana o 2. Furthermore, the identified Ana o 3 epitopes corresponding to amino acids 10–24, 13–27, 39–49, 66–70, 101–106, 107–114, and 115–122 were also screened out and chosen as the key allergenic epitopes. Results: The Ana o 2 epitopes were represented by four linear peptides, with the epitopes corresponding to amino acids 108–111, 113–119, 181–186, and 218–224. The expressed antigens were tested and confirmed using microtiter plates coated with pooled human sera from patients with cashew nut allergies or healthy controls. The peptides were also tested using phage display technology. The predicted weak and strong epitopes were synthesized as peptides. The Ana o 2 and Ana o 3 epitopes were predicted using DNAstar and PyMoL (incorporated in the Swiss-model package). Design: We predicted and experimentally confirmed cashew nut allergen antigenic peptides, which we named Ana o 2 (cupin superfamily) and Ana o 3 (prolamin superfamily). Aim: We aimed to identify the key allergenic epitopes of cashew nut proteins by correlating the phage display epitope prediction results with bioinformatics analysis. Tree nut allergies are frequently fatal and are becoming more common. Allergies to cashew nuts cause severe and systemic immune reactions. Tree nuts are one of the eight major food allergens identified by the Food and Drug Administration in the USA. Cashew nuts are a popular food source that belong to the tree nut family. Background: Cashew ( Anacardium occidentale L.) is a commercially important plant.
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