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以卵清蛋白为例,通过免疫亲和液相色谱-串联质谱法提高过敏原检测的灵敏度

Improved Sensitivity of Allergen Detection by Immunoaffinity LC-MS/MS Using Ovalbumin as a Case Study.

作者信息

Röder Martin, Wiacek Claudia, Lankamp Frauke, Kreyer Jonathan, Weber Wolfgang, Ueberham Elke

机构信息

ifp Institut für Produktqualität GmbH, Wagner-Régeny-Str. 8, 12489 Berlin, Germany.

Institute of Food Hygiene, Leipzig University, An den Tierkliniken 1, 04103 Leipzig, Germany.

出版信息

Foods. 2021 Nov 27;10(12):2932. doi: 10.3390/foods10122932.

Abstract

Food allergies are caused by severe hypersensitivity to specific food allergens such as the egg protein ovalbumin. It is therefore important to test food products for the presence of allergens to protect allergic people from accidental ingestion. For egg detection, ELISA is the only reasonable commercially available test format, although the recognition of target allergens can be affected by food processing, which may lead to false negative results. Current mass spectrometry-based detection methods may overcome this issue, but these approaches are often less sensitive. Here we combined the advantages of antibody-based and MS-based methods by developing an immunoaffinity LC-MS/MS technique to detect the common egg allergen Gal d 2. We investigated the principal functionality of this method with incurred cookie material containing whole egg powder. We found that the new method matched easily the sensitivity of egg specific ELISA tests. Further western blot experiments indicated that this strategy may be unaffected by food processing, providing an important alternative strategy for the detection and quantification of allergens in food.

摘要

食物过敏是由对特定食物过敏原(如鸡蛋蛋白卵清蛋白)的严重超敏反应引起的。因此,检测食品中是否存在过敏原对于保护过敏人群避免意外摄入至关重要。对于鸡蛋检测,酶联免疫吸附测定(ELISA)是唯一合理的市售检测形式,尽管目标过敏原的识别可能会受到食品加工的影响,这可能导致假阴性结果。目前基于质谱的检测方法可能会克服这个问题,但这些方法通常灵敏度较低。在这里,我们通过开发一种免疫亲和液相色谱-串联质谱(LC-MS/MS)技术来检测常见的鸡蛋过敏原Gal d 2,结合了基于抗体和基于质谱方法的优点。我们用含有全蛋粉的实际饼干材料研究了该方法的主要功能。我们发现新方法轻松达到了鸡蛋特异性ELISA检测的灵敏度。进一步的蛋白质印迹实验表明,该策略可能不受食品加工的影响,为食品中过敏原的检测和定量提供了一种重要的替代策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03be/8700968/6313d0da56a6/foods-10-02932-g001.jpg

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