Division of Allergology, Paul-Ehrlich-Institut, 63225 Langen, Germany.
Division of Hematology, Cell and Gene Therapy, Paul-Ehrlich-Institut, 63225 Langen, Germany.
J Agric Food Chem. 2024 Aug 14;72(32):18225-18233. doi: 10.1021/acs.jafc.4c03948. Epub 2024 Jul 30.
Allergen detection methods support food labeling and quality assessment at the allergen component level of allergen preparations used for allergy diagnosis and immunotherapy (AIT). Commonly applied enzyme-linked immunosorbent assay (ELISA) requires animal antibodies but potentially shows batch variations. We developed synthetic aptamers as alternative binders in allergen detection to meet the replacement, reduction, and refinement (3R) principle on animal protection in science. ssDNA aptamers were specifically selected against the major peanut allergen Ara h 1 and identified by next-generation sequencing. Application in various detection systems (ELISA-like assays, western blot, and surface plasmon resonance) was demonstrated. The ELISA-like assay comprised a sensitivity of 10 ng/mL Ara h 1, comparable to published antibody-based ELISA, and allowed Ara h 1 detection in various peanut flours, similar to those used in peanut AIT as well as in processed food. This ELISA-like aptamer-based assay proofs antibody-free allergen detection for food labeling or quality assessment of diagnostic and therapeutic allergen products.
过敏原检测方法支持过敏原制剂的过敏原成分水平的食品标签和质量评估，这些制剂用于过敏诊断和免疫治疗（AIT）。常用的酶联免疫吸附测定（ELISA）需要动物抗体，但可能会出现批次变化。我们开发了合成适体作为过敏原检测中的替代结合物，以满足科学中动物保护的替代、减少和优化（3R）原则。ssDNA 适体针对主要的花生过敏原 Ara h 1 进行了特异性选择，并通过下一代测序进行了鉴定。在各种检测系统（ELISA 样测定、western blot 和表面等离子体共振）中的应用得到了证明。ELISA 样测定的灵敏度为 10ng/mL Ara h 1，与已发表的基于抗体的 ELISA 相当，并允许在各种花生粉中检测 Ara h 1，类似于用于花生 AIT 以及加工食品的那些。这种基于 ELISA 样适体的测定证明了无需抗体的过敏原检测可用于食品标签或诊断和治疗性过敏原产品的质量评估。
