Chemistry Department, University of Nebraska, Lincoln, NE 68588-0304, USA.
J Pharm Biomed Anal. 2012 Oct;69:93-105. doi: 10.1016/j.jpba.2012.01.004. Epub 2012 Jan 14.
Affinity chromatography is a separation technique that has become increasingly important in work with biological samples and pharmaceutical agents. This method is based on the use of a biologically related agent as a stationary phase to selectively retain analytes or to study biological interactions. This review discusses the basic principles behind affinity chromatography and examines recent developments that have occurred in the use of this method for biomedical and pharmaceutical analysis. Techniques based on traditional affinity supports are discussed, but an emphasis is placed on methods in which affinity columns are used as part of HPLC systems or in combination with other analytical methods. General formats for affinity chromatography that are considered include step elution schemes, weak affinity chromatography, affinity extraction and affinity depletion. Specific separation techniques that are examined include lectin affinity chromatography, boronate affinity chromatography, immunoaffinity chromatography, and immobilized metal ion affinity chromatography. Approaches for the study of biological interactions by affinity chromatography are also presented, such as the measurement of equilibrium constants, rate constants, or competition and displacement effects. In addition, related developments in the use of immobilized enzyme reactors, molecularly imprinted polymers, dye ligands and aptamers are briefly considered.
亲和层析是一种分离技术,在生物样品和药物制剂的工作中变得越来越重要。该方法基于使用生物相关的试剂作为固定相来选择性地保留分析物或研究生物相互作用。本文综述了亲和层析的基本原理,并考察了该方法在生物医学和药物分析中应用的最新进展。讨论了基于传统亲和支持的技术,但重点是将亲和柱用作 HPLC 系统的一部分或与其他分析方法结合使用的方法。考虑的亲和层析的一般格式包括分步洗脱方案、弱亲和层析、亲和萃取和亲和耗尽。检查的特定分离技术包括凝集素亲和层析、硼酸亲和层析、免疫亲和层析和固定化金属离子亲和层析。还介绍了通过亲和层析研究生物相互作用的方法,例如测量平衡常数、速率常数或竞争和置换效应。此外,还简要考虑了固定化酶反应器、分子印迹聚合物、染料配体和适体在使用方面的相关进展。