Eeltink Sebastiaan, Wouters Sam, Dores-Sousa José Luís, Svec Frantisek
Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, B-1050 Brussels, Belgium.
Vrije Universiteit Brussel, Department of Chemical Engineering, Pleinlaan 2, B-1050 Brussels, Belgium.
J Chromatogr A. 2017 May 19;1498:8-21. doi: 10.1016/j.chroma.2017.01.002. Epub 2017 Jan 4.
This review focuses on the preparation of organic polymer-based monolithic stationary phases and their application in the separation of biomolecules, including antibodies, intact proteins and protein isoforms, oligonucleotides, and protein digests. Column and material properties, and the optimization of the macropore structure towards kinetic performance are also discussed. State-of-the-art liquid chromatography-mass spectrometry biomolecule separations are reviewed and practical aspects such as ion-pairing agent selection and carryover are presented. Finally, advances in comprehensive two-dimensional LC separations using monolithic columns, in particular ion-exchange×reversed-phase and reversed-phase×reversed-phase LC separations conducted at high and low pH, are shown.
本综述重点关注基于有机聚合物的整体固定相的制备及其在生物分子分离中的应用，这些生物分子包括抗体、完整蛋白质和蛋白质异构体、寡核苷酸以及蛋白质消化产物。还讨论了色谱柱和材料特性，以及针对动力学性能对大孔结构的优化。综述了当前最先进的液相色谱 - 质谱联用生物分子分离技术，并介绍了诸如离子对试剂选择和残留等实际问题。最后，展示了使用整体柱进行的全二维液相色谱分离的进展，特别是在高pH和低pH条件下进行的离子交换×反相和反相×反相液相色谱分离。
