CHROMATOGRAPHIC AUTOMATION OF IMMUNOASSAYS

描述（由申请人提供）：临床和药物化学家对血液或血清中许多药物和激素的游离或非结合部分非常感兴趣，因为这被认为代表了这些药物的活性形式。这使得这种形式作为患者诊断和治疗的理想分析工具。然而，目前还没有通用和快速的方法来测量临床样品中药物和激素的游离部分。本提案的总体目标是：(1)更好地了解药物和激素与血液或血清中的结合剂的相互作用；(2)利用这些信息开发快速、可靠的游离药物和激素检测。该项目的基本假设是，基于超快速亲和萃取和层析免疫分析技术，可以创建一类新的改进的游离药物和激素检测方法，如最近对华法林、甲状腺素、苯妥英和卡马西平等化合物的检测。未来的研究将在这些努力的基础上，考虑新的分析物和改进用于这类工作的方法。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柱；创建用于快速研究药物/激素结合和/或其结合剂的微亲和柱；并利用新方法进行药物和激素与血清制剂结合的液相动力学研究。用于测量游离药物和激素组分的改进工具将包括限制进入的亲和支持；血清蛋白超快速亲和萃取柱；自由分数测量的多维方法；而反向位移免疫分析法作为超快速亲和萃取的替代方法。这项工作将更好地了解药物和激素如何与血清因子相互作用，并导致更快，更方便的免费药物和激素测量技术。反过来，这将为临床医生提供改进的方法来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.