ABSTRACT

Affinity chromatography is an often overlooked chromatographic technique regarding receptor/ligand studies. Indeed, biologists have a tendency to focus on more recent biophysical techniques such as NMR or surface plasmon resonance methods, among others. However, in order to face challenges related to the use of membrane proteins in drug discovery, alternative and/or complementary methods are welcome. In this frame, chromatographic methods, through the affinity mode, bring new perspectives due to their relative simplicity and the great diversity of information that they can provide. Affinity chromatography methods rely on the immobilization of targets on the stationary phase and this step is crucial when dealing with membrane proteins. Different immobilization strategies with their own specificities (surface density, nonspecific interaction, complex surface chemistry of the taking or release step) were developed allowing to encompass a wide range of applications: from ligand-ranking (synthetic compounds of medium to high affinity) to fragment screening (small compounds of weak affinity), adsorption, distribution, metabolism, and excretion studies or identification of active components from natural extracts.

摘要

对于受体/配体研究，亲和色谱是一种经常被忽视的色谱技术。确实，生物学家倾向于将注意力集中在最新的生物物理技术上，例如NMR或表面等离振子共振方法。然而，为了面对与在药物开发中使用膜蛋白有关的挑战，欢迎使用替代和/或互补的方法。在这种情况下，色谱方法通过亲和模式带来了新的视角，因为它们相对简单，并且可以提供多种信息。亲和层析方法依赖于将靶标固定在固定相上，这一步骤在处理膜蛋白时至关重要。具有各自特异性的不同固定策略（表面密度，非特异性相互作用，