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。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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