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使用市售抗体实施酶联免疫吸附测定法以定量红酒中致敏性鸡蛋残留量。

Implementation of an Enzyme Linked Immunosorbent Assay for the Quantification of Allergenic Egg Residues in Red Wines Using Commercially Available Antibodies.

作者信息

Koestel Carole, Simonin Céline, Belcher Sandrine, Rösti Johannes

机构信息

Inst. of Plant Production Sciences, Swiss Federal Research Station Agroscope, Route de Duillier 50, Case Postale 1012, 1260, NYON, Switzerland.

Inst. of Food Sciences, Swiss Federal Research Station Agroscope, Route de Duillier 50, Case Postale 1012, 1260, NYON, Switzerland.

出版信息

J Food Sci. 2016 Aug;81(8):T2099-106. doi: 10.1111/1750-3841.13378. Epub 2016 Jun 29.

Abstract

Since the early 2000s, labeling of potentially allergenic food components to protect people who suffer from food allergies is compulsory in numerous industrialized countries. In Europe, milk and egg components used during the winemaking process must be indicated on the label since July 1, 2012. Several ELISA procedures have been developed to detect allergenic residues in wines. However, the complexity of the wine matrix can inhibit the immunoenzymatic reaction. The aim of this study was to implement an ELISA assay for the detection of ovalbumin in red wines using commercially available antibodies. The specificity of the acquired antibodies and the absence of cross reactivity were assessed by immunoblotting and ELISA. An ELISA assay with a LOD of 14.2 μg/L and a LOQ of 56.4 μg/L of ovalbumin in aqueous solution was obtained. Differences in ELISA signals were observed when analyzing various fining agents, although reproducible conformation of the antigen could be reached for the comparison of ovalbumin and Ovicolle. The differences between samples in terms of pH could be leveled but the inhibition of the ELISA signal, positively correlated to the tannin content of the wines, could not be suppressed. Thus, standard curves of ovalbumin in several wines were obtained by relative quantification. The control steps and the difficulties encountered presented in this study should be considered by anybody working toward the development of ELISA assays for the detection of allergenic residues in complex food matrices.

摘要

自21世纪初以来,在许多工业化国家,强制标注可能引起过敏的食品成分以保护食物过敏患者。在欧洲,自2012年7月1日起,酿酒过程中使用的牛奶和鸡蛋成分必须在标签上标明。已经开发了几种酶联免疫吸附测定(ELISA)方法来检测葡萄酒中的过敏原残留。然而,葡萄酒基质的复杂性可能会抑制免疫酶反应。本研究的目的是使用市售抗体实施一种ELISA检测方法来检测红葡萄酒中的卵清蛋白。通过免疫印迹和ELISA评估所获得抗体的特异性以及是否存在交叉反应。在水溶液中获得了一种ELISA检测方法,其卵清蛋白的检测限(LOD)为14.2μg/L,定量限(LOQ)为56.4μg/L。在分析各种澄清剂时观察到ELISA信号存在差异,尽管在比较卵清蛋白和卵黏蛋白时可以达到可重复的抗原构象。样品之间的pH差异可以消除,但与葡萄酒单宁含量呈正相关的ELISA信号抑制无法消除。因此,通过相对定量获得了几种葡萄酒中卵清蛋白的标准曲线。任何致力于开发用于检测复杂食品基质中过敏原残留的ELISA检测方法的人都应考虑本研究中提出的控制步骤和遇到的困难。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/01ff/5129457/78b25a716960/JFDS-81-T2099-g001.jpg

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