CHROMATOGRAPHIC AUTOMATION OF IMMUNOASSAYS

描述：（申请人摘要）游离或非蛋白结合形式的 许多药物和激素具有很大的临床意义， 据信代表这些化合物的生物活性部分。 这意味着这些化合物的游离浓度应该是 理想地是测量用于患者诊断或治疗。 但有 与用于此的当前方法相关的几个问题， 包括它们的速度和/或分离的单独步骤的需要， 检测游离药物或激素部分。 这项工作的目标是 开发改进的游离药物和激素的分析方法， 基于色谱的免疫测定。 这些系统的设计将基于 获得的关于模型药物和激素的速率的信息 临床样品中的结合和释放，以及它们在 固定化抗体柱。 以前的工作为我们提供了初步的 L-甲状腺素与蛋白质结合的速率常数估计值， R/S-华法林。 我们未来研究的一部分将旨在获得 更具体的信息，这些相互作用的动力学， 停流分析和使用固定化蛋白质柱。 第二 这项研究的一部分将使用计算机模拟和动力学信息 设计固定化抗体柱，用于快速提取游离 药物和激素 本研究的最后一部分将探讨使用 这些萃取柱在自动化系统的发展， 游离药物和激素的分析 在这些方法中的一些中， 提取的游离分析物将直接测定，而在其他情况下， 免疫亲和系统和柱后检测方案 荧光标记将用于定量。 以后的工作将 研究修改这些方法的方法，以具体确定 游离的药物和激素，发生在结构相似的存在下， 化合物或以不同的手性形式存在。 由于这些分析系统 发展和评估，其延伸到确定其他 游离药物或激素，加上同时测量游离和结合 溶质分数，也将被考虑。

DESCRIPTION: (applicant's abstract) The free, or non-protein bound, form of many drugs and hormones is of great clinical interest since this form is believed to represent the biologically-active fraction of these compounds. This means that the free concentrations of such compounds is what should ideally be measured for patient diagnosis or treatment. However, there are several problems associated with the current methods used for this, including their speed and/or need for separate steps for the isolation and detection of the free drug or hormone fraction. The goal of this work is to develop improved analytical methods for free drugs and hormones by using chromatographic-based immunoassays. These systems will be designed based on information obtained for model drugs and hormones regarding their rates of binding and release in clinical samples, and their rate of extraction on immobilized antibody columns. Previous work has provided us with initial estimates of rate constants for the protein-binding of L-thyroxine and R/S-warfarin. A portion of our future studies will be aimed at obtaining more specific information on the kinetics of these interactions by using stopped-flow analysis and work with immobilized protein columns. A second part of this study will use computer simulations and the kinetic information to design immobilized antibody columns for the rapid extraction of free drugs and hormones. The last portion of this study will explore the use of these extraction columns in the development of automated systems for the analysis of free drugs and hormones. In some of these methods, the extracted free analyte will be determined directly, while in others a second immunoaffinity system and a post-column detection scheme based on chemiluminescent labels will be used for quantitation. Later work will examine ways of modifying these methods for the specific determination of free drugs and hormones that occur in the presence of structurally-similar compounds or that exist in different chiral forms. As these assay systems are developed and evaluated, their extension to the determination of other free drugs or hormones, plus the simultaneous measurement of free and bound solute fractions, will also be considered.