基于单克隆抗体的酶联免疫吸附法测定金银花中绿原酸的含量。
A monoclonal antibody-based enzyme-linked immunosorbent assay for the determination of chlorogenic acid in honeysuckle.
机构信息
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China; School of Pharmacy, Linyi University, Linyi, 276000, China.
State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
出版信息
J Pharm Biomed Anal. 2018 Jan 30;148:1-5. doi: 10.1016/j.jpba.2017.09.023. Epub 2017 Sep 18.
The edible flower buds of honeysuckle are traditionally used as herbal medicine or food additives in China. Chlorogenic acid (CGA) serves as the quality control marker of honeysuckle for its high content and antipyretic property. In this paper, a specific monoclonal antibody (mAb2E2) against CGA was produced. After optimization, mAb2E2-based indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed. The concentrations of CGA producing 50% inhibition and the calibration range of the icELISA were 0.39 and 0.10-1.51ng/mL, respectively. The icELISA had cross-reactivity with 3,5-Dicaffeoylquinic acid (17.53%), and the cross-reactivity levels with other analogs were all below 5%. The average recoveries obtained by standard CGA addition to honeysuckle samples were from 88.4 to 104.8%. The icELISA was applied to CGA detection in different honeysuckle samples and the results were confirmed by high-performance liquid chromatography analysis. The correlation coefficient between the two assays was 0.97. The proposed icELISA provides a feasible analytical method for highly sensitive and specific, simple, fast, and high-throughput determination of CGA in honeysuckle samples.
金银花的食用花蕾在中国传统上被用作草药或食品添加剂。绿原酸（CGA）因其含量高和退热特性而成为金银花的质量控制标志物。本文制备了一种针对 CGA 的特异性单克隆抗体（mAb2E2）。经过优化，建立了基于 mAb2E2 的间接竞争酶联免疫吸附测定法（icELISA）。icELISA 产生 50%抑制的 CGA 浓度和校准范围分别为 0.39 和 0.10-1.51ng/mL。icELISA 与 3,5-二咖啡酰奎宁酸（17.53%）具有交叉反应性，与其他类似物的交叉反应性均低于 5%。通过向金银花样品中添加标准 CGA 获得的平均回收率在 88.4%至 104.8%之间。该 icELISA 用于不同金银花样品中的 CGA 检测，并通过高效液相色谱分析进行了验证。两种检测方法的相关性系数为 0.97。所提出的 icELISA 为金银花样品中 CGA 的高灵敏度、特异性、简单、快速和高通量检测提供了一种可行的分析方法。
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