Abstract：
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.
摘要：
过敏原检测方法支持用于过敏诊断和免疫治疗（AIT）的过敏原制剂的过敏原成分水平的食品标签和质量评估。常用的酶联免疫吸附测定（ELISA）需要动物抗体，但可能显示批次变化。我们开发了合成适体作为过敏原检测的替代结合剂，以满足替代，reduction,和科学中动物保护的细化（3R）原则。针对主要的花生过敏原Arah1特异性选择ssDNA适体，并通过下一代测序进行鉴定。在各种检测系统中的应用(ELISA样测定，westernblot,和表面等离子体共振）得到了证明。类似ELISA的测定包括10ng/mLArah1的灵敏度，与公开的基于抗体的ELISA相当，并允许在各种花生粉中检测到Arah1，与花生AIT和加工食品中使用的类似。这种基于ELISA类适体的测定证明了用于食品标签或诊断和治疗过敏原产品的质量评估的无抗体过敏原检测。
