Development of Monoclonal Antibody-based Assays for Detection and Qualification o

以青蒿素为基础的联合疗法在当前的疟疾控制和消除运动中发挥着至关重要的作用。然而，假冒和不合格青蒿素的流通极大地威胁着疟疾控制工作。不含或几乎不含有效成分的假冒青蒿素会危及生命，而不合格的药物会导致耐药性的产生。这个致命问题的规模在一些东南亚国家尤其巨大，在这些国家，市场上出售的青蒿琥酯中有高达%的产品是假冒的。为了延长治疗疟疾的青蒿素药物的寿命，需要在这些国家实施严格的药物质量控制。为此，需要快速、可靠的青蒿素检测和定量方法。传统的青蒿素定量方法往往需要昂贵的仪器和大量的技术支持，而快速定性筛选假冒青蒿素衍生物的方法不那么灵敏和可变。基于我们最近成功地利用抗青蒿素的特异性单抗开发了一种高灵敏和可靠的酶联免疫吸附试验(ELISA)，我们建议：1)利用免疫原的创新设计，开发一系列针对主要青蒿素衍生物(双氢青蒿素、蒿甲醚、青蒿醚和青蒿琥酯)的单抗；2)开发和优化青蒿素准确定量的ELISA法和快速、半定量分析抗疟疾药物中青蒿素的侧向流动试剂盒；3)使用这些新工具对东南亚三个国家的青蒿素类抗疟疾药物的药物质量进行系统的现场调查，优化酶联免疫吸附试验条件，使其可作为青蒿素及其衍生物在药代动力学研究中定量的替代工具。 这些免疫测定将为准确定量青蒿素和快速检测假冒和不合格的青蒿素衍生物药物提供便利的工具，有助于抗疟疾药物的质量控制。

Artemisinin-based combination therapies (ACTs) play an essential role in the current malaria control and elimination campaign. However, the circulation of counterfeit and substandard artemisinins greatly threatens the malaria control efforts. Fake artemisinins containing no or little active ingredients are life-threatening, while substandard drugs encourage resistance development. The scale of this 'lethal problem' is particularly huge in some Southeast Asian countries, where as much as 64% of the artesunate sold on the market was fake. To prolong the life span of artemisinin drugs for treating malaria, strict drug quality control needs to be implemented in these nations. For this purpose, fast and reliable methods of artemisinin detection and quantitation are needed. Traditional methods for quantitation of artemisinins often require expensive instruments and substantial technical support, whereas the rapid qualitative method for screening of fake artemisinin derivatives is less sensitive and variable. Based on our recent success of developing a highly sensitive and reliable enzyme-linked immunosorbent assay (ELISA) using a specific monoclonal antibody against artemisinins, we propose to 1) develop a series of monoclonal antibodies with specificities for the major artemisinin derivatives (dihydroartemisinin, artemether, arteether, and artesunate) using an innovative design of immunogens, 2) develop and optimize an ELISA for accurate quantification of artemisinins, and a lateral flow dipstick assay for rapid, semi-quantitative analysis of artemisinins in antimalarial drugs, 3) perform systematic field surveys of the drug quality of artemisinin-based antimalarial medicines in three Southeast Asian countries using these new tools, and 4) optimize the ELISA conditions so that it can be used as an alternative tool for quantification of artemisinins and its derivatives in pharmacokinetic studies. These immunoassays will offer convenient tools for both accurate quantification of artemisinins and rapid detection of counterfeit and substandard artemisinin derivative drugs, contributing to antimalarial drug quality control.