Applications of Monoclonal Antibodies Against Natural Compounds for Functional Analysis of Crude Drugs
Applications of Monoclonal Antibodies Against Natural Compounds for Functional Analysis of Crude Drugs
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DOI：
10.1007/s40495-020-00224-7
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发表时间：
2020-07
影响因子：
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通讯作者：
T. Uto;Shunsuke Fujii;S. Sakamoto;Tomoe Ohta;Y. Shoyama
中科院分区：
文献类型：
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作者：
T. Uto;Shunsuke Fujii;S. Sakamoto;Tomoe Ohta;Y. Shoyama
综述目的本研究总结了针对生物活性天然化合物的单克隆抗体（MAb）在生药功能分析中的应用。最新研究成果迄今为止，我们已经建立了40多种针对天然化合物的单克隆抗体，并建立了酶联免疫吸附测定（ELISA）。使用抗 HT MAb 通过 ELISA 测定细胞内三尖杉酯碱 (HT) 及其衍生物的积累，这些化合物的摄取水平与其各自的抗增殖活性相符。 Eastern blotting 是一种膜上定量分析系统，使用单克隆抗体对抗天然化合物，也可用于生物样品中的药代动力学研究。通过东部印迹法在接受 GC 治疗的大鼠血清中检测到甘草甜素 (GC)。此外，使用针对 3-单葡萄糖醛酸基-甘草次酸 (3-MGA) 的 MAb 进行东部印迹分析可以确定由过量摄入 GC 引起的假性醛固酮增多症的潜在致病因子。与 MAb 结合的免疫亲和柱使得制备含有除目标化合物之外的所有成分的提取物变得可行。使用通过与抗 GC MAb 偶联的免疫亲和柱制备的去除 GC 的甘草提取物进行的细胞研究揭示了 GC 在甘草提取物中的潜在功能。此外，单克隆抗体还用于确定天然化合物的细胞定位和靶分子。使用 MAb 和 LC-MS/MS 进行的分析相结合表明，马兜铃酸 I (AA-I) 是一种天然存在的肾毒素和致癌物质，以肾细胞中的 α-actinin-4 为靶标。针对天然化合物的 MAb 是确定粗提物中天然化合物的真实药理功能、细胞定位和分子靶点的有用工具。
Purpose of ReviewThis study summarizes the applications of monoclonal antibodies (MAbs) against bioactive natural compounds for the functional analysis of crude drugs.Recent FindingsTo date, we have established more than 40 kinds of MAbs against natural compounds and set up enzyme-linked immunosorbent assay (ELISA). The intracellular accumulation of harringtonine (HT) and its derivatives into cells was determined by ELISA using anti-HT MAb, and the uptake levels of these compounds conformed to their respective antiproliferative activities. Eastern blotting is an on-membrane quantitative analytical system that uses MAbs against natural compounds and it can also be used in a biological sample for pharmacokinetics study. Glycyrrhizin (GC) was detected in the serum of rats treated with GC by eastern blotting. Additionally, eastern blotting using MAb against 3-monoglucuronyl-glycyrrhetinic acid (3-MGA) could determine a potential causative agent of pseudoaldosteronism induced by excess intake of GC. The immunoaffinity column coupled with MAb made preparing an extract containing all components feasible except the target compound. Cell-based studies using GC-removed licorice extract, which was prepared by the immunoaffinity column conjugated with anti-GC MAb, revealed the potential functions of GC in licorice extract. Moreover, MAbs were used in determining cellular localization and target molecules of natural compounds. The combination of analysis using MAb and LC-MS/MS showed that aristolochic acid I (AA-I), a naturally occurring nephrotoxin and carcinogen, targets α-actinin-4 in the kidney cells.SummaryMAbs against natural compounds are useful tools to determine real pharmacological functions, cellular localization, and molecular targets of natural compounds in crude extracts.