CHROMATOGRAPHIC AUTOMATION OF IMMUNOASSAYS

在临床样本中，药物或药物的游离或非蛋白质结合部分 人们对荷尔蒙非常感兴趣，因为它代表着化合物的 具有生物活性的形式。这使得对这一分数的测量 非常适合患者诊断和治疗。但目前的分析方法 因为这些测量(通常基于免疫分析)是缓慢的，分娩- 密集，不能由现有的自动化系统执行。这是 由于必须测定的低浓度，以及需要 在分析前分离分析物的结合部分和游离部分。在……里面 此外，分离过程中结合分析物解离的影响 过程没有被很好地理解。这项研究的目的是获得一个 更好地理解这种分离，并开发新的、自动化的 免费药物/荷尔蒙测量系统。解离效果将是 通过检测两种模型化合物甲状腺激素的蛋白质结合进行了研究 和皮质醇。用于自动检测这些药物的免费药物/激素的系统 基于高效免疫亲和力的化合物将被开发 层析和化学发光检测(HPIAC/CL)。在这种方法中， 固定化抗体柱将用于快速提取游离分析物 从样品中提取，然后使用化学发光检测分析物 吖啶酸酯标签。 这项工作的第一部分将研究相互作用动力学 甲状腺激素和皮质醇及其结合蛋白。这将通过以下方式完成 研究其结合蛋白AS的保留和扩宽 它们通过含有固定的甲状腺激素或皮质醇的柱子。 目的是为了更好地了解解离效应。 免费药物/激素分析，并提供将这些影响降至最低的数据 在HPIAC/CL系统中。第二部分将研究所涉及的过程。 将分析物吸附到固定化抗体柱上。我们的目标是 为了确定需要什么最佳的层析条件 使用HPIAC/CL自动进行免费药物/激素免疫分析。这将是 使用计算机模拟和使用模型系统进行研究， 刀豆蛋白A与荧光和非荧光糖的结合。这个 最后一节将使用以前研究中的数据来设计系统 根据HPIAC/CL测定游离甲状腺激素和皮质醇。一个 将这种方法与目前的方法进行比较，以确定 它的潜在优势和局限性。该系统在中国的应用 然后还将考虑测量其他药物/荷尔蒙。

In clinical samples the free, or non-protein bound, fraction of a drug or hormone is of great interest since it represents the compound's biologically-active form. This makes the measurement of this fraction ideal for patient diagnosis and treatment. But current analytical methods for these measurements (typically based on immunoassays) are slow, labor- intensive, and cannot be performed by existing automated systems. This is due to the low concentrations which must be determined and the need to separate the bound and free fractions of the analyte prior to analysis. In addition, the effects of bound analyte dissociation during the separation process are not well understood. The aim of this study is to obtain a better understanding of this dissociation and to develop new, automated systems for free drug/hormone measurements. Dissociation effects will be studied by examining the protein binding of two model compounds, thyroxine and cortisol. Systems for automating free drug/hormone assays for these compounds will be developed based on high-performance immunoaffinity chromatography and chemiluminescent detection (HPIAC/CL). In this method, an immobilized antibody column will be used to rapidly extract free analyte from a sample, followed by analyte detection using chemiluminescent acridinium ester labels. The first section of this work will examine the interaction kinetics of thyroxine and cortisol with their binding proteins. This will be done by studying the retention and band-broadening of their binding proteins as they pass through columns containing immobilized thyroxine or cortisol. The goal is to obtain a better understanding of dissociation effects in free drug/hormone assays and to provide data for minimizing these effects in HPIAC/CL systems. The second section will study the processes involved in the adsorption of analytes to immobilized antibody columns. The goal is to determine what optimum chromatographic conditions are needed for automating free drug/hormone immunoassays using HPIAC/CL. This will be studied using computer simulations and work with a model system, the binding of concanavalin A to fluorescent and nonfluorescent sugars. The last section will use data from the previous studies to design systems based on HPIAC/CL for free thyroxine and cortisol measurements. A comparison of this approach to current methods will be made to determine its potential advantages and limitations. The use of this system in measuring other drugs/hormones will then also be considered.