Development of validated sandwich ELISA for detecting peanut allergen Ara h 3 in food

• Published in: Food chemistry Vol. 445; p. 138757
• Main Authors: Lin, Jia-Fong, Chang, Kee-Lung, Hsieh, Bau-Shan, Hu, Yu-Chen, Huang, Edward S., Yu, Hsu-Sheng
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.07.2024
• Subjects: Allergen; allergens; analytical methods; Antibody; Ara h 3; asthma; ELISA; enzyme-linked immunosorbent assay; food chemistry; Peanut; peanuts; Purification; Peanut; Purification; Antibody; Allergen; Ara h 3; ELISA
• ISSN: 0308-8146; 1873-7072; 1873-7072
• DOI: 10.1016/j.foodchem.2024.138757

•The recovery of the ELISA was 0.63%-4.08%.•The intra-day and inter-day precision were 0.63–4.08% and 3.56%-5.78%.•The LOD and LOQ were 0.023 μg/mL and 0.033 μg/mL.•The method showed no cross-reactivity with soybeans, cashew nuts and sesame.•Ara h 3 can be detected in different food matrices containing peanuts. Peanut is an important food that can cause food allergies, often leading to moderate and severe allergic symptoms such as skin rashes, asthma, and even anaphylactic shock.Research indicates that Ara h 3 is one of the major peanut allergen. In order to establish a simple analytical method for detecting Ara h 3, we developed a sandwich enzyme-linked immunosorbent assay (ELISA) with antibodies that were induced from purified Ara h 3. The experimental results showed that the purified Ara h 3 had good purity, and we successfully prepared capture and detection antibodies. The method established in this study exhibited high specificity and did not cross-react with soybeans, cashew nuts, and sesame. For validation, including precision, recovery and sensitivity were in good condition. We also detected the Ara h 3 in peanut related foods. Overall, the ELISA developed in this study is a reliable method for Ara h 3 detection.