建立酶联免疫吸附测定法和滴板法快速分析葡萄果实中反式白藜芦醇。
Development of an enzyme-linked immunosorbent assay and a dipstick assay for the rapid analysis of trans-resveratrol in grape berries.
机构信息
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China; College of Science, China Agricultural University, Beijing 100193, China.
Tropic Crops Genetic Resources Institute, Chinese Academy of Tropic Agricultural Sciences, Danzhou, Hainan 571737, China.
出版信息
Food Chem. 2019 Sep 1;291:132-138. doi: 10.1016/j.foodchem.2019.04.023. Epub 2019 Apr 6.
trans-Resveratrol, a natural polyphenol present in grapes, has many beneficial effects to human health. However, measurement of trans-resveratrol is technically complicated and time-consuming. In the present study, we obtained a sensitive and specific monoclonal antibody (mAb), namely 3C9, against trans-resveratrol. An indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed, with 50% inhibitory concentration and working range of 1.0 ng/mL and 0.19-4.9 ng/mL, respectively. A lateral flow immunoassay (LFIA) based on 3C9 was also developed for the semiquantitative detection of trans-resveratrol in an indicator range of 50-100 ng/mL. Average recoveries of trans-resveratrol spiked in red and green grape berries samples were respectively 88-107% and 83-102% by icELISA. The icELISA and LFIA were applied for determination of trans-resveratrol in grape berries. The results were highly consistent with those determined by high-performance liquid chromatography analysis (R = 0.9997). Therefore, we conclude that the immunoassay methods are suitable for the large-scale screening of trans-resveratrol in grape quality breeding.
反式白藜芦醇是葡萄中存在的一种天然多酚，对人体健康有许多有益的影响。然而，反式白藜芦醇的测量技术复杂且耗时。在本研究中，我们获得了一种针对反式白藜芦醇的敏感和特异的单克隆抗体（mAb），即 3C9。建立了间接竞争酶联免疫吸附测定（icELISA），其 50%抑制浓度和工作范围分别为 1.0ng/mL 和 0.19-4.9ng/mL。还基于 3C9 开发了侧向流动免疫分析（LFIA），用于在 50-100ng/mL 的指示范围内对半定量检测反式白藜芦醇。icELISA 测定的红、绿葡萄浆果中添加的反式白藜芦醇的平均回收率分别为 88-107%和 83-102%。icELISA 和 LFIA 用于测定葡萄浆果中的反式白藜芦醇。结果与高效液相色谱分析（R = 0.9997）高度一致。因此，我们得出结论，免疫测定方法适用于葡萄品质育种中反式白藜芦醇的大规模筛选。
Zotero 插件