Leurs Ulrike, Mistarz Ulrik H, Rand Kasper D
Department of Pharmacy, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
Department of Pharmacy, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen, Denmark.
Eur J Pharm Biopharm. 2015 Jun;93:95-109. doi: 10.1016/j.ejpb.2015.03.012. Epub 2015 Mar 17.
Protein pharmaceuticals are the fastest growing class of novel therapeutic agents, and have been a major research and development focus in the (bio)pharmaceutical industry. Due to their large size and structural diversity, biopharmaceuticals represent a formidable challenge regarding analysis and characterization compared to traditional small molecule drugs. Any changes to the primary, secondary, tertiary or quaternary structure of a protein can potentially impact its function, efficacy and safety. The analysis and characterization of (structural) protein heterogeneity is therefore of utmost importance. Mass spectrometry has evolved as a powerful tool for the characterization of both primary and higher order structures of protein pharmaceuticals. Furthermore, the chemical and physical stability of protein drugs, as well as their pharmacokinetics are nowadays routinely determined by mass spectrometry. Here we review current techniques in primary, secondary and tertiary structure analysis of proteins by mass spectrometry. An overview of established top-down and bottom-up protein analyses will be given, and in particular the use of advanced technologies such as hydrogen/deuterium exchange mass spectrometry (HDX-MS) for higher-order structure analysis will be discussed. Modification and degradation pathways of protein drugs and their detection by mass spectrometry will be described, as well as the growing use of mass spectrometry to assist protein design and biopharmaceutical development.
蛋白质药物是新型治疗剂中增长最快的类别，并且一直是（生物）制药行业主要的研发重点。由于其尺寸大且结构多样，与传统小分子药物相比，生物制药在分析和表征方面面临巨大挑战。蛋白质的一级、二级、三级或四级结构的任何变化都可能潜在地影响其功能、功效和安全性。因此，（结构）蛋白质异质性的分析和表征至关重要。质谱已发展成为表征蛋白质药物一级和高级结构的强大工具。此外，蛋白质药物的化学和物理稳定性以及它们的药代动力学如今通常通过质谱来确定。在此，我们综述了通过质谱分析蛋白质一级、二级和三级结构的当前技术。将给出已确立的自上而下和自下而上蛋白质分析的概述，尤其将讨论使用诸如氢/氘交换质谱（HDX-MS）等先进技术进行高级结构分析。将描述蛋白质药物的修饰和降解途径及其通过质谱的检测，以及质谱在辅助蛋白质设计和生物制药开发方面日益增加的应用。
