CHROMATOGRAPHIC AUTOMATION OF IMMUNOASSAYS

描述(由申请人提供)：血液或血清中许多药物和激素的自由或非结合部分引起临床和药物化学家的极大兴趣，因为这被认为代表了这些药物的活性形式。这使得该表格成为患者诊断和治疗的理想分析工具。然而，目前还没有一种通用的、快速的方法来测量临床样品中药物和激素的游离部分。这项建议的总体目标是1)更好地了解药物和激素与其在血液或血清中的结合剂之间的相互作用，以及2)利用这些信息开发快速、可靠的免费药物和激素检测方法。该项目的基本假设是，可以基于超快亲和提取和层析免疫分析技术创建一类新的改进的游离药物和激素分析，就像最近对华法林、甲状腺素、苯妥英和卡马西平等化合物所证明的那样。未来的研究将在这些努力的基础上，考虑新的分析物和用于这类工作的方法的改进。有待探索的进展包括开发用于研究药物与血清脂蛋白结合的亲和色谱柱；创建用于快速研究药物/激素结合和/或其结合剂包埋的微亲和色谱柱；以及使用新方法研究药物和激素与血清试剂结合的溶液相动力学。将创建用于测量游离药物和激素部分的改进工具，包括限制访问的亲和支撑物；用于超快亲和提取的血清蛋白质柱；用于测量游离部分的多维方法；以及作为超快亲和提取的替代方法的反向置换免疫分析。这项工作将更好地了解药物和荷尔蒙如何与血清试剂相互作用，并产生更快、更方便的免费药物和荷尔蒙测量技术。反过来，这将为临床医生提供改进的方法，研究这些化合物在体内的行为和相互作用。

DESCRIPTION (provided by applicant): The free, or non-bound, fraction of many drugs and hormones in blood or serum is of great interest to clinical and pharmaceutical chemists since this is believed to represent the active form of such agents. This makes this form ideal as an analytical tool for patient diagnosis and treatment. However, there is currently no general and fast approach for measuring the free fractions of drugs and hormones in clinical samples. The overall goals of this proposal are 1) to obtain a better understanding of drug and hormone interactions with their binding agents in blood or serum and 2) to use this information to develop fast, reliable free drug and hormone assays. The underlying hypothesis of this project is that a new class of improved free drug and hormone assays can be created based on the techniques of ultrafast affinity extraction and chromatographic immunoassays, as recently demonstrated for compounds such as warfarin, thyroxine, phenytoin and carbamazepine. Future studies will build on these efforts by considering new analytes and improvements in the methods used for such work. Advances to be explored will include the development of affinity columns for studying the binding of drugs to serum lipoproteins; the creation of microaffinity columns for rapid studies of drug/hormone binding and/or the entrapment of their binding agents; and the use of new methods for solution-phase kinetic studies of drug and hormone binding to serum agents. Improved tools to be created for the measurement of free drug and hormone fractions will include restricted access affinity supports; serum protein columns for ultrafast affinity extractions; multidimensional methods for free fraction measurements; and the reverse displacement immunoassay as an alternative to ultrafast affinity extraction. This work will provide a better understanding of how drugs and hormones interact with serum agents and result in faster, more convenient techniques for free drug and hormone measurements. This, in turn, will provide clinicians with improved methods for studying how such compounds behave and interact in the body.