A highly specific competitive enzyme-linked immunosorbent assay (ELISA) protocol has been developed to identify and classify almond products based on differential proteomic analysis. We applied two-dimensional electrophoresis to compare the differences between almond and apricot kernels to search for almond-specific proteins. The amino acid of apricot Pru-1 was sequenced and aligned to almond Pru-1. One peptide, RQGRQQGRQQQEEGR, which exists in almond but not in apricot, was used as hapten to prepare monoclonal antibody against almond Pru-1. An optimized ELISA method was established using this antibody. The assay did not exhibit cross-reactivity with the tested apricot kernels and other edible plant seeds. The limit of detection (LOD) was 2.5-100μg/g based on different food samples. The recoveries of fortified samples at levels of twofold and eightfold LOD ranged from 82% to 96%. The coefficients of variation were less than 13.0%. Using 7M urea as extracting solution, the heat-treated protein loss ratios were 2%, 5% and 15% under pasteurization (65°C for 30min), baking (150°C for 30min) and autoclaved sterilization (120°C for 15min), respectively.

中文翻译：

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基于差异蛋白质组学分析的杏仁副产物高特异性定量方法。

已经开发出一种高度特异性的竞争性酶联免疫吸附测定（ELISA）方案，以基于差异蛋白质组学分析对杏仁产品进行鉴定和分类。我们应用二维电泳来比较杏仁和杏仁之间的差异，以搜索杏仁特异的蛋白质。对杏Pru-1的氨基酸进行测序，并与杏仁Pru-1进行比对。一种存在于杏仁中但不存在于杏中的肽RQGRQQGRQQQEEGR被用作半抗原，以制备针对杏仁Pru-1的单克隆抗体。使用该抗体建立了优化的ELISA方法。该测定与被测的杏仁和其他可食用植物种子没有交叉反应。根据不同食品样品的检测限（LOD）为2.5-100μg/ g。在两倍和八倍LOD水平下，强化样品的回收率在82％至96％之间。变异系数小于13.0％。使用7M尿素作为提取液，在巴氏杀菌（65°C 30分钟），烘烤（150°C 30分钟）和高压灭菌（120°C 15分钟）下热处理的蛋白质损失率分别为2％，5％和15％。 ）， 分别。