[Functional analysis of bioactive natural compounds using monoclonal antibodies against natural compounds].
[Functional analysis of bioactive natural compounds using monoclonal antibodies against natural compounds].
复制标题
[使用针对天然化合物的单克隆抗体对生物活性天然化合物进行功能分析]。
DOI：
10.1248/yakushi.14-00178
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
T. Uto
中科院分区：
文献类型：
--
作者：
T. Uto
由于人们对草药的健康益处越来越感兴趣，草药最近受到了极大的重视。因此，有必要进一步阐明这些天然化合物的功能和作用机理。我们实验室已经建立了30多种针对具有生物活性的天然化合物的单抗。此外，我们还开发了天然化合物的高灵敏度测量系统，如酶联免疫吸附试验(ELISA)和使用单抗的East blotting。为了扩大这些单抗在天然化合物功能分析中的应用，我们建立了一种从植物提取物中分离目标化合物的新方法，使用与抗天然化合物单抗结合的免疫亲和柱。通过单抗偶联免疫亲和柱的一步纯化，我们成功地制备了包含除目标化合物外的所有组分的敲除(KO)提取物，用作半抗原。此外，我们还考察了KO提取物的药理作用，以确定该生物活性化合物在植物提取物中的确切作用。为了证实单抗的另一个有益用途，我们利用单抗通过免疫细胞化学(ICC)和Western blotting研究了天然化合物的细胞定位和靶分子。我们的结果表明，单抗清楚地决定了天然化合物的细胞定位和靶分子。这些方法可能使确定草药中具有生物活性的天然化合物的潜在功能和靶分子成为可能。
Herbal medicines have recently attracted much importance owing to the rising interest in their health benefits. Hence, further elucidation of the functions and mechanisms of these natural compounds is necessary. Our laboratory has established more than 30 kinds of monoclonal antibodies (MAbs) against bioactive natural compounds. Moreover, we have developed highly sensitive measurement systems for natural compounds, such as enzyme-linked immunosorbent assay (ELISA) and eastern blotting using MAbs. To expand the application of these MAbs to the functional analysis of natural compounds, we established a new approach for the isolation of the target compound from plant extracts using an immunoaffinity column conjugated with an anti-natural compound MAb. Through one-step purification using a MAb-conjugated immunoaffinity column, we have succeeded in preparing a knockout (KO) extract containing all components except the target compound, used as a hapten. Furthermore, we examined the pharmacological effects of the KO extract to identify the precise roles of the bioactive compound in the plant extract. To confirm another beneficial use of MAbs, we investigated the cellular localization and target molecules of natural compounds by immunocytochemistry (ICC) and Western blotting using MAbs. Our results demonstrated that MAbs clearly determined the cellular localization and target molecules of the natural compounds. These approaches may make it possible to determine the potential functions and target molecules of bioactive natural compounds in herbal medicines.