Establishment of monoclonal antibodies of BW10kDa distinguish it from Fag e 2 related to anaphylaxis

IF 4.1 Q2 FOOD SCIENCE & TECHNOLOGY Food Chemistry Molecular Sciences Pub Date : 2024-05-26 DOI:10.1016/j.fochms.2024.100207

Masaya Kimura , Rie Satoh , Reiko Teshima

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Abstract

BW10kDa, which is a buckwheat (BW) allergen, belongs to the 2S-albumin protein family, akin to Fag e 2. Detailed analyses of BW10kDa were lacking until this study. Herein, we conducted these analyses using monoclonal antibodies (mAbs) to recombinant BW10kDa (rBW10kDa). We successfully generated anti-rBW10kDa mAbs capable of distinguishing between Fag e 2 and BW10kDa. These mAbs were categorised into two types (type 1 and type 2) based on their reactivity to BW plant seed extracts in western blot analyses. Type 1 mAbs revealed two bands (15 kDa and 10 kDa), while type 2 mAbs showed a single band (15 kDa). Spot analyses using these mAbs confirmed that type 1 mAbs recognised epitopes near the C-terminal region, with the 10 kDa band representing the C-terminal subunit cleaved by protease. The mAbs targeting rBW10kDa enabled to assess the concentration of BW10kDa in wild type and also in diagnostic buckwheat extracts.

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建立 BW10kDa 的单克隆抗体，将其与过敏性休克相关的 Fag e 2 区分开来

BW10kDa是一种荞麦（BW）过敏原，属于2S-白蛋白家族，与Fag e 2相似。在本研究之前，还缺乏对 BW10kDa 的详细分析。在此，我们使用重组 BW10kDa（rBW10kDa）的单克隆抗体（mAbs）进行了这些分析。我们成功生成了能够区分 Fag e 2 和 BW10kDa 的抗 rBW10kDa mAbs。根据这些 mAbs 在 Western 印迹分析中对 BW 植物种子提取物的反应性，将其分为两种类型（1 型和 2 型）。1 型 mAbs 显示两条带（15 kDa 和 10 kDa），而 2 型 mAbs 显示一条带（15 kDa）。使用这些 mAbs 进行的斑点分析证实，1 型 mAbs 可识别 C 端附近的表位，10 kDa 条带代表被蛋白酶裂解的 C 端亚基。靶向 rBW10kDa 的 mAbs 能够评估野生型和诊断荞麦提取物中 BW10kDa 的浓度。

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来源期刊

Food Chemistry Molecular Sciences Agricultural and Biological Sciences-Food Science

CiteScore

6.00

自引率

0.00%

发文量

审稿时长

82 days

期刊介绍： Food Chemistry: Molecular Sciences is one of three companion journals to the highly respected Food Chemistry. Food Chemistry: Molecular Sciences is an open access journal publishing research advancing the theory and practice of molecular sciences of foods. The types of articles considered are original research articles, analytical methods, comprehensive reviews and commentaries. Topics include: Molecular sciences relating to major and minor components of food (nutrients and bioactives) and their physiological, sensory, flavour, and microbiological aspects; data must be sufficient to demonstrate relevance to foods and as consumed by humans Changes in molecular composition or structure in foods occurring or induced during growth, distribution and processing (industrial or domestic) or as a result of human metabolism Quality, safety, authenticity and traceability of foods and packaging materials Valorisation of food waste arising from processing and exploitation of by-products Molecular sciences of additives, contaminants including agro-chemicals, together with their metabolism, food fate and benefit: risk to human health Novel analytical and computational (bioinformatics) methods related to foods as consumed, nutrients and bioactives, sensory, metabolic fate, and origins of foods. Articles must be concerned with new or novel methods or novel uses and must be applied to real-world samples to demonstrate robustness. Those dealing with significant improvements to existing methods or foods and commodities from different regions, and re-use of existing data will be considered, provided authors can establish sufficient originality.