Alterations in protein structure may have profound effects on biological function. Analytical techniques that permit characterization of proteins while maintaining their conformational and functional state are crucial for studying changes in the higher order structure of proteins and for establishing structure–function relationships. Coupling of native protein separations with mass spectrometry is emerging rapidly as a powerful approach to study these aspects in a reliable, fast and straightforward way. This Review presents the available native separation modes for proteins, covers practical considerations on the hyphenation of these separations with mass spectrometry and highlights the involvement of affinity-based separations to simultaneously obtain structural and functional information of proteins. The impact of these approaches is emphasized by selected applications addressing biomedical and biopharmaceutical research questions.

蛋白质结构的改变可能对生物功能产生深远影响。允许表征蛋白质同时保持其构象和功能状态的分析技术对于研究蛋白质高级结构的变化和建立结构-功能关系至关重要。天然蛋白质分离与质谱联用正在迅速成为一种以可靠、快速和直接的方式研究这些方面的强大方法。本综述介绍了可用的蛋白质天然分离模式，涵盖了这些分离与质谱联用的实际考虑，并强调了基于亲和力的分离的参与，以同时获取蛋白质的结构和功能信息。