Jeong Kyoung Yong, Park Kyung Hee, Lee Jae Hyun, Park Jung Won
Department of Internal Medicine, Institute of Allergy, Yonsei University College of Medicine, Seoul, Korea.
Allergy Asthma Immunol Res. 2017 Sep;9(5):417-422. doi: 10.4168/aair.2017.9.5.417.
Buckwheat is a major cause of anaphylaxis, and Fag e 3 is the key major allergen in buckwheat. However, an immunoassay system for the quantification of Fag e 3 has yet to be developed.
We developed a 2-site enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies (mAbs) produced against recombinant Fag e 3. We applied this ELISA to quantify native Fag e 3 in total buckwheat extract.
Four clones of mAbs were produced, and all recognized vicilin allergens not only from buckwheat, but also from peanut and walnut. However, the ELISA using these antibodies was only able to quantify Fag e 3 in the total extract after addition of 1% sodium dodecyl sulphate (SDS) and heating, which facilitated dissociation of the allergen. The detection limit of the developed 2-site ELISA was 0.8 μg/mL. The measurement of Fag e 3 in the total extract of buckwheat showed that approximately 12% of protein in total buckwheat extract was Fag e 3.
We have developed an ELISA system for the quantification of the group 3 buckwheat allergen, Fag e 3, specifically. This assay will be useful for standardization of buckwheat allergens and monitoring of buckwheat contamination in foods.
荞麦是过敏反应的主要诱因，而荞麦过敏原Fag e 3是荞麦中的关键主要过敏原。然而，用于定量Fag e 3的免疫分析系统尚未开发出来。
我们利用针对重组Fag e 3产生的单克隆抗体（mAb）开发了一种双位点酶联免疫吸附测定（ELISA）。我们应用这种ELISA来定量全荞麦提取物中的天然Fag e 3。
产生了4个单克隆抗体克隆，所有克隆均识别不仅来自荞麦，还来自花生和核桃的豌豆球蛋白过敏原。然而，使用这些抗体的ELISA仅在添加1%十二烷基硫酸钠（SDS）并加热后才能定量全提取物中的Fag e 3，这有助于过敏原的解离。所开发的双位点ELISA的检测限为0.8μg/mL。对荞麦全提取物中Fag e 3的测量表明，荞麦全提取物中约12%的蛋白质是Fag e 3。
我们专门开发了一种ELISA系统，用于定量第3组荞麦过敏原Fag e 3。该测定法将有助于荞麦过敏原的标准化以及食品中荞麦污染的监测。
