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

以青蒿素为基础的联合疗法(ACTs)在当前疟疾控制和治疗中发挥着至关重要的作用 淘汰赛。然而，假冒和不合格青蒿素的流通极大地威胁到 疟疾控制的努力。不含或几乎不含青蒿素成分的假青蒿素会危及生命， 而不合格的药物则会鼓励耐药性的发展。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NAfions中实施严格的药品质量控制。为此，需要一种快速、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灵敏度较低，变量较少。基于我们最近成功地利用针对青蒿素的特异性单抗开发了一种高灵敏度和可靠的酶联免疫吸附试验(ELISA)，我们建议1)利用创新的免疫原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)开发和优化一种准确测定青蒿素含量的ELISA和一种用于快速、半定量分析抗疟疾药物青蒿素的横向流动扩散法，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品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在药代动力学研究中定量的替代工具。 这些免疫分析方法为准确定量检测青蒿素类药物和快速检测假冒伪劣青蒿素类药物提供了方便的工具，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.