Dinh Ngoc Phuoc, Cam Quach Minh, Nguyen Anh Mai, Shchukarev Andrei, Irgum Knut
Department of Chemistry, Umeå University, Umeå, Sweden.
J Sep Sci. 2009 Aug;32(15-16):2556-64. doi: 10.1002/jssc.200900243.
Macroporous epoxy-based monoliths prepared by emulsion polymerization have been modified for use in ion exchange chromatography (IEC) of proteins. Strong anion exchange functionality was established by iodomethane quaternization of tertiary amine present on the monolith surface as a part of the polymer backbone. The modification pathway to cation exchange materials was via incorporation of glycidyl methacrylate (GMA) brushes which were coated using atom transfer radical polymerization (ATRP). Strong (SO(3)(-)) and weak (COO(-)) cation exchange groups were thereafter introduced onto the GMA-grafted monoliths by reactions with sodium hydrogen sulfite and iminodiacetic acid, respectively. Grafting was confirmed by XPS, gravimetric measurement, and chromatographic behavior of the modified materials toward model proteins. In incubation experiments the proteins were recovered quantitatively with no obvious signs of unfolding after contact with the stationary phase for >2 h. Chromatographic assessments on the functionalized columns as well as problems associated with flow-through modification by ATRP are discussed.
通过乳液聚合制备的大孔环氧基整体柱已被改性，用于蛋白质的离子交换色谱（IEC）。通过对作为聚合物主链一部分存在于整体柱表面的叔胺进行碘甲烷季铵化，建立了强阴离子交换功能。阳离子交换材料的改性途径是通过使用原子转移自由基聚合（ATRP）涂覆甲基丙烯酸缩水甘油酯（GMA）刷。此后，分别通过与亚硫酸氢钠和亚氨基二乙酸反应，将强（SO(3)(-)）和弱（COO(-)）阳离子交换基团引入到接枝了GMA的整体柱上。通过XPS、重量测量以及改性材料对模型蛋白质的色谱行为证实了接枝。在孵育实验中，蛋白质在与固定相接触>2小时后被定量回收，且没有明显的展开迹象。讨论了功能化柱的色谱评估以及与ATRP流通改性相关的问题。
