THERAPEUTIC DRUG MONITORING BY CAPILLARY IMMUNOPHORESIS

我们提出的研究的主要目标是整合一个在线 毛细管电泳免疫检测技术在多残留分析中应用 结构相关的药物及其代谢物在体液中。 毛细管免疫电泳作为一种混合分析技术， 电泳分离的分析能力和免疫选择性， 微量治疗药物的鉴定和定量 复杂样品基质中的药物。类似于二维的 分离系统，毛细管电泳作为第一维， 根据药物分析物的大小、电特性和 电荷和疏水性。免疫反应，作为第二个维度 分离利用抗体的交叉反应性来识别 具有共同结构元件（表位）的药物分析物。 为了实现我们的研究目标，毛细管的三个技术特点， 免疫电泳必须解决：（i）分离模式的选择 以及电渗透控制对分离分辨率的影响 毛细管电泳中的药物分析物（ii）定量 所施加电场强度与毛细管尺寸的相关性，以及 溶液离子强度对分析物转移有效性的影响， 在毛细管后免疫反应器中将药物分析物与抗体混合， 和（iii）抗原（药物分析物）-抗体的直接感测 通过监测半抗原的荧光偏振反应， 荧光素缀合物（示踪剂）在竞争性免疫测定形式中的应用。到 进一步简化药物监测程序，减少与健康有关的 我们的长期目标是展示 毛细管免疫电泳同时检测几种 通常滥用的或治疗性的药物。 毛细管免疫电泳作为治疗药物的革命性方法 监测显示出选择性量子跳跃的显著前景， 灵敏度和速度超过目前的分析方法。的 毛细管免疫电泳的发展及其成功依赖于 在不同领域建立了许多技术里程碑，包括 本项目中概述的以及未来的整合 微机械加工和阵列检测技术。

The main goal of our proposed research is to integrate an on-line immunodetection with capillary electrophoresis for multiresidue analysis of structurally related drugs and their metabolites in body fluids. Capillary immunophoresis as a hybrid analytical technique exhibits the analytical power of electrophoretic separation and immunoselectivity for the identification and quantification of trace amounts of therapeutic drugs in complex sample matrices. In analogy to a two-dimensional separation system, capillary electrophoresis as the first dimension, resolves drug analytes based on differences in their size, electrical charge, and hydrophobicity. Immunoreaction, as the second dimension of separation utilizes the antibody's cross-reactivity for the recognition of drug analytes with a common structural element (epitope). To achieve our research goal, three technical features of capillary immunophoresis have to be addressed: (i) the selection of separation mode and the effect of electroosmotic control on the separation resolution of drug analytes in capillary electrophoresis (ii) the quantitative dependence of applied electric field strengths capillary dimensions, and solution ionic strengths on the effectiveness of analyte transfer and mixing of drug analytes with antibodies in a post-capillary immunoreactor, and (iii) the direct sensing of the antigen (drug analyte)-antibody reaction by monitoring the fluorescence polarization of the hapten- fluorescein conjugate (tracer) in a competitive immunoassay format. To further simplify the drug monitoring procedures and reduce health-related costs, our long-term objective is to demonstrate the potential of capillary immunophoresis for the simultaneous detection of several commonly abused or therapeutic drugs in a single biological fluid sample. Capillary immunophoresis as a revolutionary method for therapeutic drug monitoring shows significant promise for quantum jumps in selectivity, sensitivity, and speed over the current analytical methods. The development of capillary immunophoresis and its success relies on establishing many technical milestones in diverse fields including those outlined in this project as well as the future integration of micromachining and array detection techniques.