Department of Chemistry, University of Nebraska, Lincoln, NE 68588-0304, USA.
Analyst. 2018 Jan 15;143(2):374-391. doi: 10.1039/c7an01469d.
The last few decades have witnessed the development of many high-performance separation methods that use biologically related binding agents. The combination of HPLC with these binding agents results in a technique known as high performance affinity chromatography (HPAC). This review will discuss the general principles of HPAC and related techniques, with an emphasis on their use for the analysis of biological compounds and pharmaceutical agents. Various types of binding agents for these methods will be considered, including antibodies, immunoglobulin-binding proteins, aptamers, enzymes, lectins, transport proteins, lipids, and carbohydrates. Formats that will be discussed for these methods range from the direct detection of an analyte to indirect detection based on chromatographic immunoassays, as well as schemes based on analyte extraction or depletion, post-column detection, and multi-column systems. The use of biological agents in HPLC for chiral separations will also be considered, along with the use of HPAC as a tool to screen or study biological interactions. Various examples will be presented to illustrate these approaches and their applications in fields such as biochemistry, clinical chemistry, and pharmaceutical research.
过去几十年见证了许多使用生物相关结合剂的高性能分离方法的发展。将 HPLC 与这些结合剂相结合，产生了一种称为高效亲和色谱（HPAC）的技术。本文综述将讨论 HPAC 及相关技术的一般原理，重点介绍它们在分析生物化合物和药物制剂中的应用。将考虑用于这些方法的各种类型的结合剂，包括抗体、免疫球蛋白结合蛋白、适体、酶、凝集素、转运蛋白、脂质和碳水化合物。本文将讨论这些方法的各种形式，从直接检测分析物到基于色谱免疫测定的间接检测，以及基于分析物提取或耗尽、柱后检测和多柱系统的方案。本文还将考虑在 HPLC 中使用生物试剂进行手性分离，以及将 HPAC 用作筛选或研究生物相互作用的工具。将提供各种实例来说明这些方法及其在生物化学、临床化学和药物研究等领域的应用。
