Hage David S
Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE.
Clin Chem. 2017 Jun;63(6):1083-1093. doi: 10.1373/clinchem.2016.262253. Epub 2017 Apr 10.
The interactions between biochemical and chemical agents in the body are important in many clinical processes. Affinity chromatography and high-performance affinity chromatography (HPAC), in which a column contains an immobilized biologically related binding agent, are 2 methods that can be used to study these interactions.
This review presents various approaches that can be used in affinity chromatography and HPAC to characterize the strength or rate of a biological interaction, the number and types of sites that are involved in this process, and the interactions between multiple solutes for the same binding agent. A number of applications for these methods are examined, with an emphasis on recent developments and high-performance affinity methods. These applications include the use of these techniques for fundamental studies of biological interactions, high-throughput screening of drugs, work with modified proteins, tools for personalized medicine, and studies of drug-drug competition for a common binding agent.
The wide range of formats and detection methods that can be used with affinity chromatography and HPAC for examining biological interactions makes these tools attractive for various clinical and pharmaceutical applications. Future directions in the development of small-scale columns and the coupling of these methods with other techniques, such as mass spectrometry or other separation methods, should continue to increase the flexibility and ease with which these approaches can be used in work involving clinical or pharmaceutical samples.
体内生化试剂与化学试剂之间的相互作用在许多临床过程中都很重要。亲和色谱法和高效亲和色谱法(HPAC),其中柱中含有固定化的生物相关结合剂,是可用于研究这些相互作用的两种方法。
本综述介绍了可用于亲和色谱法和高效亲和色谱法的各种方法,以表征生物相互作用的强度或速率、参与该过程的位点数量和类型,以及同一结合剂与多种溶质之间的相互作用。研究了这些方法的一些应用,重点是近期的发展和高效亲和方法。这些应用包括将这些技术用于生物相互作用的基础研究、药物的高通量筛选、修饰蛋白的研究、个性化医疗工具以及针对共同结合剂的药物-药物竞争研究。
可与亲和色谱法和高效亲和色谱法一起用于研究生物相互作用的多种形式和检测方法,使这些工具在各种临床和制药应用中具有吸引力。小型柱开发的未来方向以及这些方法与其他技术(如质谱或其他分离方法)的联用,应会继续提高这些方法在涉及临床或制药样品的工作中的灵活性和易用性。