Central Laboratory, Shaanxi Provincial People's Hospital, Research Center of Cell Immunological Engineering and Technology of Shaanxi Province, Xi'an, Shaanxi 710068, P.R. China.
School of Pharmacy, Lanzhou University, Lanzhou, Gansu 730000, P.R. China.
Mol Med Rep. 2019 Aug;20(2):959-966. doi: 10.3892/mmr.2019.10342. Epub 2019 Jun 5.
The use of camptothecin and its analogues has increased in clinical settings and in agriculture. Therefore, camptothecins and their derivatives, metabolites and degradation products are frequently found in the environment. Therefore, it is important to develop an ELISA for the quantification of camptothecins in human plasma, plants, animal tissues and other matrices. The present study developed a novel competitive indirect ELISA for camptothecin using a monoclonal antibody (MAb). In total, two haptens and various carrier proteins were tested to select the most suitable immunogen for the production of MAbs against camptothecin. Hapten 1 conjugated with keyhole limpet hemocyanin was selected for the preparation of MAb 5A3, and was used to establish a competitive indirect ELISA for camptothecin. A total of three derivatives of camptothecin used in clinical practice were examined. Topotecan showed an IC50 value of 0.68 µg/ml with a detection limit of 0.19 µg/ml, belotecan showed an IC50 value of 0.87 µg/ml with a detection limit of 0.22 µg/ml and irinotecan showed an IC50 value of 2.85 µg/ml with a detection limit of 0.47 µg/ml. The cross‑reactivity results suggested that the assay developed in the present study possessed a high sensitivity to camptothecin. Therefore, this immunoassay technique may be suitable for monitoring the levels of camptothecin in compound analysis, clinical applications, and analyses of food and environmental samples.
喜树碱及其类似物在临床和农业领域的应用日益增多。因此，喜树碱及其衍生物、代谢物和降解产物经常在环境中被发现。因此，开发一种用于定量检测人血浆、植物、动物组织和其他基质中喜树碱的 ELISA 方法非常重要。本研究使用单克隆抗体（MAb）开发了一种新型的竞争性间接 ELISA 方法来检测喜树碱。总共测试了两种半抗原和各种载体蛋白，以选择最适合用于制备抗喜树碱 MAb 的免疫原。将半抗原 1 与血蓝蛋白偶联，用于制备 MAb 5A3，并用于建立竞争性间接 ELISA 方法来检测喜树碱。本研究还检测了三种临床使用的喜树碱衍生物。拓扑替康的 IC50 值为 0.68µg/ml，检测限为 0.19µg/ml；贝洛替康的 IC50 值为 0.87µg/ml，检测限为 0.22µg/ml；伊立替康的 IC50 值为 2.85µg/ml，检测限为 0.47µg/ml。交叉反应性结果表明，本研究中建立的测定法对喜树碱具有高灵敏度。因此，这种免疫分析技术可能适用于监测化合物分析、临床应用和食品与环境样品中喜树碱的水平。
