College of Agronomy and Biotechnology, China Agricultural University, Beijing, China.
PLoS One. 2013 Nov 13;8(11):e79154. doi: 10.1371/journal.pone.0079154. eCollection 2013.
Artemether is one of the artemisinin derivatives that are active ingredients in antimalarial drugs. Counterfeit and substandard antimalarial drugs have become a serious problem, which demands reliable analytical tools and implementation of strict regulation of drug quality. Structural similarity among artemisinin analogs is a challenge to develop immunoassays that are specific to artemisinin derivatives. To produce specific antibodies to artemether, we used microbial fermentation of artemether to obtain 9-hydroxyartemether, which was subsequently used to prepare a 9-O-succinylartemether hapten for conjugation with ovalbumin as the immunogen. A monoclonal antibody (mAb), designated as 2G12E1, was produced with high specificity to artemether. 2G12E1 showed low cross reactivities to dihydroartemisinin, artemisinin, artesunate and other major antimalarial drugs. An indirect competitive enzyme linked immunosorbent assay (icELISA) developed showed a concentration causing 50% of inhibition for artemether as 3.7 ng mL⁻¹ and a working range of 0.7-19 ng mL⁻¹. The icELISA was applied for determination of artemether content in different commercial drugs and the results were comparable to those determined by high-performance liquid chromatography analysis. In comparison with reported broad cross activity of anti-artemisinin mAbs, the most notable advantage of the 2G12E1-based ELISA is its high specificity to artemether only.
蒿甲醚是青蒿素类药物的有效成分之一。假冒伪劣抗疟药物已成为一个严重问题，这就需要可靠的分析工具，并实施严格的药品质量监管。青蒿素类似物的结构相似性给青蒿素衍生物的免疫分析带来了挑战。为了生产针对蒿甲醚的特异性抗体，我们使用微生物发酵蒿甲醚获得 9-羟基蒿甲醚，随后将其用于制备与卵清蛋白偶联的 9-O-琥珀酰蒿甲醚半抗原作为免疫原。我们制备了一种特异性识别蒿甲醚的单克隆抗体（mAb），命名为 2G12E1。2G12E1 对双氢青蒿素、青蒿素、青蒿琥酯和其他主要抗疟药物的交叉反应性较低。建立的间接竞争酶联免疫吸附测定（icELISA）显示，蒿甲醚的半数抑制浓度（IC50）为 3.7ng mL⁻¹，工作范围为 0.7-19ng mL⁻¹。该 icELISA 用于测定不同商业药物中的蒿甲醚含量，结果与高效液相色谱分析结果相当。与报道的广谱抗青蒿素 mAb 相比，基于 2G12E1 的 ELISA 的最显著优势是其对蒿甲醚的高特异性。
