Wu Longjiang, Dang Mei, Wu Rao, Isah Murtala Bindawa, Zhang Xiaoying
Chinese-German Joint Institute for Natural Product Research, Shaanxi International Cooperation Demonstration Base, Shaanxi University of Technology, Hanzhong, China.
Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore, Singapore.
Front Nutr. 2024 Aug 21;11:1439287. doi: 10.3389/fnut.2024.1439287. eCollection 2024.
3-caffeoylquinic acid (3-CQA), a member of the chlorogenic acid family, possesses diverse pharmacological properties, such as scavenging, antioxidant, and antiapoptotic activity, rendering substantial value to alimentary consumables and therapeutic substances. However, the pervasiveness of non-standard practices, notably the misuse and abuse of indigenous botanicals, coupled with the inherent susceptibility of 3-CQA to degradation under light and heat exposure, engenders discernible disparateness in the quality profiles of the same kinds of herbs. Consequently, precise quantification of 3-CQA becomes imperative.
In this context, an artificial antigen was synthesized as a specific conjugate of 3-CQA and bovine serum albumin (3-CQA-BSA), followed by the generation of a monoclonal antibody (mAb) against the conjugate. Through optimization, a mAb-based indirect competitive chemiluminescence enzyme immunoassay (ic-CLEIA) was developed.
It demonstrated an IC and the calibration range of 2.97 ng/mL and 0.64-13.75 ng/mL, respectively, outperforming the conventional enzyme-linked immunosorbent assay (ELISA). Notably, the ic-CLEIA displayed 10.71% cross-reactivity with 3,5-dicaffeoylquinic acid, alongside minimal cross-reactivity toward other isomeric counterparts and analogs. Validation experiments on herbs and Chinese patent medicines using ic-CLEIA, confirmed by high-performance liquid chromatography (HPLC) analysis, revealed a robust correlation coefficient of 0.9667 between the two modalities.
These findings unequivocally demonstrated that the proposed ic-CLEIA represents a viable and reliable analytical method for 3-CQA determination. This method holds significant potential for ensuring the quality control and therapeutic efficacy germane to herbs and patent medicines, spanning diverse therapeutic milieus and applications.
3-咖啡酰奎宁酸（3-CQA）是绿原酸家族的一员，具有多种药理特性，如清除、抗氧化和抗凋亡活性，对食品和治疗物质具有重要价值。然而，非标准做法的普遍存在，特别是对本土植物药的滥用和误用，再加上3-CQA在光照和热暴露下易降解的固有特性，导致同类草药的质量特征存在明显差异。因此，精确定量3-CQA变得至关重要。
在此背景下，合成了一种人工抗原，即3-CQA与牛血清白蛋白的特异性偶联物（3-CQA-BSA），随后产生了针对该偶联物的单克隆抗体（mAb）。通过优化，开发了一种基于mAb的间接竞争化学发光酶免疫分析方法（ic-CLEIA）。
该方法的IC50和校准范围分别为2.97 ng/mL和0.64 - 13.75 ng/mL，优于传统的酶联免疫吸附测定（ELISA）。值得注意的是，ic-CLEIA与3,5-二咖啡酰奎宁酸的交叉反应率为10.71%，对其他异构体和类似物的交叉反应率极低。使用ic-CLEIA对草药和中成药进行的验证实验，经高效液相色谱（HPLC）分析证实，两种方法之间的相关系数高达0.9667。
这些结果明确表明，所提出的ic-CLEIA是一种可行且可靠的3-CQA测定分析方法。该方法在确保草药和中成药的质量控制和治疗效果方面具有巨大潜力，适用于各种治疗环境和应用。
