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基于免疫磁珠脂质体荧光免疫分析的无麸质食品中麦胶蛋白污染的灵敏检测和定量

Sensitive detection and quantification of gliadin contamination in gluten-free food with immunomagnetic beads based liposomal fluorescence immunoassay.

机构信息

Department of Food Science and Biotechnology, National Chung Hsing University, Taichung, Taiwan, ROC.

出版信息

Anal Chim Acta. 2013 Jul 17;787:246-53. doi: 10.1016/j.aca.2013.05.014. Epub 2013 May 27.

Abstract

Gliadin from wheat is a common food allergen that can induce baker's asthma, wheat-dependent exercise-induced anaphylaxis, atopic dermatitis, and celiac disease. This gliadin assay focuses on rapidly screen and check for gluten contamination in raw materials and in the gluten-free food production process, not only for wheat-sensitive patients but also for the industries producing gluten-free foodstuffs. The developed assay incorporates the use of anti-gliadin antibody-conjugated immunomagnetic beads (IMBs) to capture the gliadin in samples and fluorescent dyes-loaded immunoliposomal nanovesicles (IMLNs) to produce and enhance the detection signal. Hence, a sandwich complex is formed as "IMBs-gliadin-IMLNs". Experimental results indicate that this detection platform exhibits good sensitivity for gliadin with a detection limit as low as 0.6 μg mL(-1) of gliadin; as the polyclonal antibody showed slight cross-reactions with barley and rye. Excellent recovery rates were found ranging from 83.5 to 102.6% as testing the spiked samples. Moreover, the CV (%) of intra- and inter-assay of this developed assay are 4.8-10.6% and 3.5-9.9%, respectively. Based on a parallel analysis of twenty food samples, the results of this developed assay provide a good consistency with those of an AOAC-approved ELISA kit without any false-negative results. The proposed assay method is thus a highly promising alternative method for detecting the contamination of gliadin in the food industry.

摘要

小麦醇溶蛋白是一种常见的食物过敏原,可引起烘焙师哮喘、小麦依赖运动诱发的过敏反应、特应性皮炎和乳糜泻。该醇溶蛋白检测法主要用于快速筛选和检测原材料及无麸质食品生产过程中的麸质污染,不仅适用于小麦敏感患者,也适用于生产无麸质食品的行业。该检测法结合使用抗醇溶蛋白抗体偶联免疫磁珠(IMB)来捕获样品中的醇溶蛋白和负载荧光染料的免疫脂质体纳米囊泡(IMLN)以产生和增强检测信号。因此,形成了“IMB-醇溶蛋白-IMLN”的三明治复合物。实验结果表明,该检测平台对醇溶蛋白具有良好的灵敏度,检测限低至 0.6μg mL(-1)的醇溶蛋白;由于多克隆抗体与大麦和黑麦略有交叉反应。在测试加标样品时,发现回收率非常高,范围在 83.5%至 102.6%之间。此外,该检测法的内、间测定的 CV(%)分别为 4.8-10.6%和 3.5-9.9%。通过对二十个食品样品的平行分析,该检测法的结果与经 AOAC 批准的 ELISA 试剂盒的结果非常一致,没有出现任何假阴性结果。因此,该检测方法是一种很有前途的替代方法,可用于检测食品工业中的醇溶蛋白污染。

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