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

以青蒿素为基础的联合疗法在目前的疟疾控制中发挥着重要作用， 消除运动。然而，假冒伪劣青蒿素的流通极大地威胁着 控制疟疾的努力。不含或含有少量acfive成分的假青蒿素会危及生命， 而不合格的药物会导致抗药性的产生。这个“致命问题”的规模在一些东南亚国家尤其巨大，在那里，市场上销售的青蒿琥酯多达64%是假的。为延长青蒿素类抗疟药的使用寿命，需要对这些国家的青蒿素类抗疟药进行严格的质量控制。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，而快速定性方法用于筛选假青蒿素衍生物的灵敏度和可变性较低。基于我们最近成功地建立了一种高灵敏度和可靠的青蒿素单克隆抗体酶联免疫吸附试验（ELISA），我们提出：1）研制一系列对主要青蒿素衍生物具有特异性的单克隆抗体（双氢青蒿素、蒿甲醚、蒿乙醚和青蒿琥酯），2）建立并优化了青蒿素的ELISA定量方法和抗疟药中青蒿素的快速半定量分析方法，3）使用这些新工具，在三个东南亚国家对青蒿素类抗疟药物的药物质量进行系统的实地调查，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药代动力学研究中定量分析的替代工具。 这些免疫分析方法为青蒿素类药物的准确定量和快速检测伪、劣药提供了方便的手段，有助于抗疟药的质量控制。

Artemisinin-based combination therapies (ACTs) play an essenfial 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 acfive ingredients are life-threatening, while substandard drugs encourage resistance development. The scale of this 'lethal problem' is particulariy 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 nafions. For this purpose, fast and reliable methods of artemisinin detection and quanfitation 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 sensifive and variable. Based on our recent success of developing a highly sensifive and reliable enzyme-linked immunosorbent assay (ELISA) using a specific monoclonal anfibody 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 dipsfick assay for rapid, semi-quanfitafive analysis of artemisinins in anfimalarial drugs, 3) perform systematic field surveys of the drug quality of artemisinin-based anfimalarial 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 quanfificafion of artemisinins and rapid detecfion of counterfeit and substandard artemisinin derivative drugs, contribufing to anfimalarial drug quality control.