Preparation of Anti-Glycyrrhetinic Acid Monoclonal Antibody for Application in an Indirect Competitive Enzyme-Linked Immunosorbent Assay
Preparation of Anti-Glycyrrhetinic Acid Monoclonal Antibody for Application in an Indirect Competitive Enzyme-Linked Immunosorbent Assay
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DOI：
10.1080/00032719.2017.1370598
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发表时间：
2018-02
影响因子：
2
通讯作者：
Shunsuke Fujii;T. Uto;Shuichi Nomura;Y. Shoyama
中科院分区：
文献类型：
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作者：
Shunsuke Fujii;T. Uto;Shuichi Nomura;Y. Shoyama
摘要 甘草次酸是甘草甜素的主要代谢产物，是甘草根的主要成分之一，被认为是甘草中的药理活性物质之一。通过融合小鼠骨髓瘤细胞和脾细胞产生了一种新的杂交瘤细胞系，命名为 G-2A6，并使用甘草次酸-匙孔血蓝蛋白进行免疫，产生抗甘草次酸的单克隆抗体 (mAb)。使用抗甘草次酸单克隆抗体，我们尝试开发一种简单、快速、高灵敏度的间接竞争性酶联免疫吸附测定（icELISA）。开发的 icELISA 范围为 3.91 至 125 ng/mL，变异系数低（小于 5%），并且表明甘草粉中掺入甘草次酸的回收率很高（平均值 = 101.76%）。此外，icELISA 可以通过简单的预处理来测定加有甘草次酸的人血清中的甘草次酸浓度，这表明使用抗甘草次酸单克隆抗体开发的 ELISA 系统将被证明是在甘草植物和甘草次酸分析等各个领域中测定甘草次酸的有效且有用的工具。 甘草次酸、甘草甜素和/或甘草类药物给药过程中甘草次酸的药代动力学研究。
ABSTRACT Glycyrrhetinic acid is a major metabolite of glycyrrhizin, which is one of the main components of licorice roots and is considered to be one of the pharmacologically active substances in licorice. A new hybridoma cell line, named G-2A6, was generated by fusing mouse myeloma cells and splenocytes, which were immunized using glycyrrhetinic-acid–keyhole limpet hemocyanin to produce a monoclonal antibody (mAb) against glycyrrhetinic acid. Using the anti-glycyrrhetinic acid mAb, we attempted to develop a simple, rapid, and highly sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA). The developed icELISA had a range from 3.91 to 125 ng/mL with low coefficients of variation (less than 5%) and demonstrated a high recovery rate of glycyrrhetinic acid spiked into licorice powder (average = 101.76%). In addition, the icELISA could determine the glycyrrhetinic acid concentration in glycyrrhetinic-acid-spiked human serum with simple pretreatment, which suggests that the developed ELISA system using anti-glycyrrhetinic acid mAb would prove to be an effective and useful tool for determining glycyrrhetinic acid in various fields such as the analysis of Glycyrrhiza plants and pharmacokinetic studies of glycyrrhetinic acid during the administration of glycyrrhetinic acid, glycyrrhizin, and/or licorice-based medical agents.