Abstract In this study, a cashew allergen, Ana o 3 (2S albumin), was used as a protein marker to develop two sensitive and specific sandwich ELISAs (denoted as CAS-ELISA-2 and Ano3-ELISA-1) with the limits of detection (LOD, determined as a concentration that yield 20% of maximum colour development) at 0.04 and 0.06 mg protein kg−1, respectively. The ELISAs were highly specific to cashew nut with very low cross-reactivity with pistachio, pecan, almond, peanut, and hazelnut. Assay validation on the model cookies observed lower cashew protein recovery of 40.2–44.9% with the CAS–ELISA–2 assay and 25.2–79.8% with the Ano3–ELISA–1 assay. Both assays exhibited a limit of quantitation (LOQ) of 40 mg protein kg−1 cookies. While for the model chocolate, the acceptable protein recoveries were obtained with both CAS–ELISA–2 (74.0–87.7%) and Ano3–ELISA–1 (62.8–136.2%). The LOQ of 4 mg protein kg−1 chocolate were achieved.

中文翻译：

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开发特异性针对 2S 白蛋白 (Ana o 3) 的灵敏夹心 ELISA，作为预包装食品中腰果残留检测的稳定蛋白质标记

摘要 在本研究中，以腰果过敏原 Ana o 3（2S 白蛋白）为蛋白质标记物，开发了两种灵敏且特异的夹心 ELISA（表示为 CAS-ELISA-2 和 Ano3-ELISA-1），其限值为检测（LOD，确定为产生最大显色的 20% 的浓度）分别为 0.04 和 0.06 毫克蛋白质 kg-1。ELISA 对腰果具有高度特异性，与开心果、山核桃、杏仁、花生和榛子的交叉反应性非常低。对模型饼干的测定验证观察到，CAS-ELISA-2 测定的腰果蛋白回收率较低，为 40.2-44.9%，Ano3-ELISA-1 测定为 25.2-79.8%。两种测定都显示出 40 毫克蛋白质 kg-1 饼干的定量限 (LOQ)。而对于模型巧克力，使用 CAS-ELISA-2 (74.0-87.7%) 和 Ano3-ELISA-1 (62.0%) 均获得了可接受的蛋白质回收率。8–136.2%）。达到了 4 mg 蛋白质 kg-1 巧克力的 LOQ。