Pandey, A K and Varshney, R K and Sudini, H K and Pandey, M K (2019) An Improved Enzyme-Linked Immunosorbent Assay (ELISA) Based Protocol Using Seeds for Detection of Five Major Peanut Allergens Ara h 1, Ara h 2, Ara h 3, Ara h 6, and Ara h 8. Frontiers in Nutrition (TSI), 6 (68). pp. 1-11. ISSN 2296-861X
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Abstract
Peanut allergy is an important health concern among many individuals. As there is no effective treatment to peanut allergy, continuous monitoring of peanut-based products, and their sources is essential. Precise detection of peanut allergens is key for identification and development of improved peanut varieties with minimum or no allergens in addition to estimating the levels in peanut-based products available in food chain. The antibody based ELISA protocol along with sample preparation was standardized for Ara h 1, Ara h 2, Ara h 3, Ara h 6, and Ara h 8 to estimate their quantities in peanut seeds. Three different dilutions were optimized to precisely quantify target allergen proteins in peanut seeds such as Ara h 1 (1/1,000, 1/2,000, and 1/4,000), Ara h 2 and Ara h 3 (1/5,000, 1/10,000, and 1/20,000), Ara h 6 (1/40,000, 1/80,000, and 1/1,60,000), and Ara h 8 (1/10, 1/20, and 1/40). These dilutions were finalized for each allergen based on the accuracy of detection by achieving <20% coefficient of variation in three technical replicates. This protocol captured wide variation of allergen proteins in selected peanut genotypes for Ara h 1 (77–46,106 μg/g), Ara h 2 (265–5,426 μg/g), Ara h 3 (382–12,676 μg/g), Ara h 6 (949–43,375 μg/g), and Ara h 8 (0.385–6 μg/g). The assay is sensitive and reliable in precise detection of five major peanut allergens in seeds. Deployment of such protocol allows screening of large scale germplasm and breeding lines while developing peanut varieties with minimum allergenicity to ensure food safety.
| Item Type: | Article |
|---|---|
| Divisions: | Research Program : Asia Research Program : Genetic Gains |
| CRP: | CGIAR Research Program on Grain Legumes and Dryland Cereals (GLDC) |
| Uncontrolled Keywords: | protocol, peanut allergens, Ara h 1, Ara h 2, Ara h 3, Ara h 6, Ara h 8, ELISA, Food allergies, food safety, groundnut |
| Subjects: | Mandate crops > Groundnut |
| Depositing User: | Mr Ramesh K |
| Date Deposited: | 21 Aug 2019 05:24 |
| Last Modified: | 21 Aug 2019 05:24 |
| URI: | http://oar.icrisat.org/id/eprint/11257 |
| Official URL: | https://doi.org/10.3389/fnut.2019.00068 |
| Projects: | UNSPECIFIED |
| Funders: | UNSPECIFIED |
| Acknowledgement: | Authors are thankful to Dr. Hari D. Upadhyaya for providing seeds of genotypes included in the study. This work has been undertaken as part of the CGIAR Research Program on Grain Legumes and Dryland Cereals (GLDC). ICRISAT is a member of CGIAR Consortium. |
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