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基于电感耦合等离子体质谱检测的适体分析超灵敏检测β-伴大豆球蛋白食物过敏原。

Ultrasensitive determination of β-conglutin food allergen by means an aptamer assay based on inductively coupled plasma mass spectrometry detection.

机构信息

University of Alicante, Department of Analytical Chemistry, Nutrition and Food Sciences, PO Box 99, 03080, Alicante, Spain.

INTERFIBIO Consolidated Research Group, Department of Chemical Engineering, Universitat Rovira i Virgili, 43007, Tarragona, Spain.

出版信息

Anal Chim Acta. 2023 Apr 29;1252:341042. doi: 10.1016/j.aca.2023.341042. Epub 2023 Mar 3.

Abstract

The overall objective of this work is the evaluation of different competitive aptamer assays based on inductively coupled plasma mass spectrometry (ICP-MS) detection for the determination of β-conglutin (food protein allergen from lupin) in flour samples. To this end, two competitive aptamer assay schemes were developed using either thiolated aptamers chemisorbed onto gold nanoparticles (AuNPs) or biotinylated aptamers linked to streptavidin-AuNPs. The influence of ICP-MS detection mode (i.e., conventional vs single particle) on assay performance was explored. In the case of the thiolated aptamer, the limit of detection (LoD) obtained using the single particle mode was improved 2-fold as compared to the LoD provided by the conventional mode. With regards to the biotinylated aptamer, the use of the conventional mode provided a 5-fold improvement of LoD as compared to that obtained for the single particle one. Using the optimized conditions, the best LoD of 2 pM was obtained with the biotinylated aptamer operating with conventional ICP-MS detection. When compared to previous reports using the same aptamer in a competitive assay, the developed method significantly improved the LoD by at least an order of magnitude. Different flour samples containing lupin were successfully analyzed according to European Conformity guidelines for the analysis of food contaminants.

摘要

本工作的总体目标是评估基于电感耦合等离子体质谱(ICP-MS)检测的不同竞争适体分析方法,用于测定面粉样品中的β-羽扇豆球蛋白(来自羽扇豆的食物蛋白过敏原)。为此,开发了两种竞争适体分析方案,分别使用化学吸附在金纳米粒子(AuNPs)上的巯基化适体或连接到链霉亲和素-AuNPs 的生物素化适体。探讨了 ICP-MS 检测模式(即常规模式与单颗粒模式)对分析性能的影响。对于巯基化适体,与常规模式相比,单颗粒模式下获得的检测限(LoD)提高了 2 倍。对于生物素化适体,与单颗粒模式相比,常规模式的 LoD 提高了 5 倍。在优化条件下,使用常规 ICP-MS 检测的生物素化适体获得了最佳 LoD 为 2 pM。与使用相同适体在竞争性分析中报告的先前研究相比,开发的方法至少将 LoD 提高了一个数量级。根据欧洲符合性准则对食品污染物进行分析,成功分析了不同含有羽扇豆的面粉样品。

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