Frostell Asa, Vinterbäck Lena, Sjöbom Hans
GE Healthcare Bio-Sciences AB, Uppsala, Sweden.
Methods Mol Biol. 2013;1008:139-65. doi: 10.1007/978-1-62703-398-5_6.
Surface plasmon resonance (SPR) biosensor technology has become an important tool for drug discovery and basic research. SPR instruments are used for a wide variety of applications including determining the binding kinetics and affinity of an interaction, specificity studies, screening, assay development as well as concentration measurements. The interacting molecules may be proteins, peptides, lipids, viruses, nucleic acids, or small organic molecules such as fragments or drug candidates. The ease with which real time information can be obtained has changed many customer workflows in both antibody and small molecule/fragment interaction analysis, from label based and affinity/IC50 based workflows towards a label free and kinetic based workflow. This chapter focuses on applications for drug discovery, and outlines the experimental design for screening and selection of small molecules from a focused library. Also, determination of kinetics and/or affinity constants of selected ligands, using established SPR methodology is described, together with potential issues during assay development, running of the assay, and results interpretation.
表面等离子体共振（SPR）生物传感器技术已成为药物发现和基础研究的重要工具。SPR仪器可用于多种应用，包括确定相互作用的结合动力学和亲和力、特异性研究、筛选、分析方法开发以及浓度测量。相互作用的分子可能是蛋白质、肽、脂质、病毒、核酸或小分子，如片段或候选药物。能够轻松获取实时信息，改变了抗体和小分子/片段相互作用分析中许多客户的工作流程，从基于标记和基于亲和力/IC50的工作流程转向基于无标记和动力学的工作流程。本章重点介绍药物发现的应用，并概述从聚焦文库中筛选和选择小分子的实验设计。此外，还描述了使用既定的SPR方法测定所选配体的动力学和/或亲和力常数，以及分析方法开发、分析运行和结果解释过程中的潜在问题。
