Department of Food Science and Human Nutrition, Faculty of Agriculture, Ryukoku University.
Department of Food and Nutrition, Kyoto Women's University.
J Nutr Sci Vitaminol (Tokyo). 2022;68(3):221-227. doi: 10.3177/jnsv.68.221.
In this study, monoclonal antibodies against two major fruit allergens-gibberellin-regulated protein (GRP) and lipid transfer protein (LTP)-were established. Sandwich enzyme-linked immunosorbent assays (ELISAs) for the quantification of peach GRP and LTP were constructed using these antibodies. Both ELISAs reacted with the respective antigens when heated at 100ºC for 20 min, but not when reduced with sodium sulfite, indicating that GRP and LTP are heat-stable, while disulfide bonds play an important role in their native steric structures. GRP and LTP in peaches and peach-containing foods were quantified by these ELISAs. In both cases, there were few differences among peach cultivars normally available on the market; however, concentrations were higher when the peach was ripe. GRP was localized in the pulp of the peach, while LTP was present in the peel. They could be quantified in peach-containing beverages, as well as in dried and canned peaches. GRP in Japanese apricots could also be determined using this ELISA, as its amino acid sequence is the same as that of peach GRP. Then, high concentrations of GRP were detected in umeboshi, a traditional Japanese pickled apricot. Peach leaves were found to have a high LTP content, accordingly, LTP was also observed in lotions containing peach leaf extract. The ability to quantitatively detect GRP and LTP in this study will, therefore, contribute to the improvement of component-resolved diagnoses and quality of life in patients allergic to peaches.
在这项研究中，建立了针对两种主要水果过敏原——赤霉素调节蛋白（GRP）和脂转移蛋白（LTP）的单克隆抗体。使用这些抗体构建了用于定量检测桃 GRP 和 LTP 的夹心酶联免疫吸附测定法（ELISA）。当在 100°C 加热 20 分钟时，两种 ELISA 都与各自的抗原发生反应，但当用亚硫酸钠还原时则不发生反应，这表明 GRP 和 LTP 是热稳定的，而二硫键在其天然空间结构中起着重要作用。通过这些 ELISA 定量检测了桃和含桃食品中的 GRP 和 LTP。在这两种情况下，市场上通常可买到的桃品种之间差异不大；但是，当桃子成熟时，浓度会更高。GRP 定位在桃的果肉中，而 LTP 存在于果皮中。它们可以在含桃的饮料中，以及在干桃和罐装桃中进行定量检测。使用这种 ELISA 还可以测定日本李脯中的 GRP，因为它的氨基酸序列与桃 GRP 相同。然后，在传统的日本腌制李脯 umeboshi 中检测到 GRP 的高浓度。发现桃叶含有高含量的 LTP，因此，在含有桃叶提取物的乳液中也观察到了 LTP。因此，本研究中定量检测 GRP 和 LTP 的能力将有助于改善对桃过敏患者的成分分辨诊断和生活质量。
