Qu Jian-Bo, Xu Yu-Liang, Liu Jun-Yi, Zeng Jing-Bin, Chen Yan-Li, Zhou Wei-Qing, Liu Jian-Guo
State Key Laboratory of Heavy Oil Processing, Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, PR China.
State Key Laboratory of Heavy Oil Processing, Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, PR China.
J Chromatogr A. 2016 Apr 8;1441:60-7. doi: 10.1016/j.chroma.2016.02.072. Epub 2016 Mar 2.
Dual thermo- and pH-responsive chromatography has been proposed using poly(N-isopropylacrylamide-co-butyl methacrylate-co-N,N-dimethylaminopropyl acrylamide) (P(NIPAM-co-BMA-co-DMAPAAM)) brushes grafted gigaporous polystyrene microspheres (GPM) as matrix. Atom transfer radical polymerization (ATRP) initiator was first coupled onto GPM through Friedel-Crafts acylation with 2-bromoisobutyryl bromide. The dual-responsive polymer brushes were then grafted onto GPM via surface-initiated ATRP. The surface composition, gigaporous structure, protein adsorption and dual-responsive chromatographic properties of the matrix (GPM-P(NIPAM-co-BMA-co-DMAPAAM) were characterized in detail. Results showed that GPM were successfully grafted with thermoresponsive cationic polymer brushes and that the gigaporous structure was well maintained. A column packed with GPM-P(NIPAM-co-BMA-co-DMAPAAM presented low backpressure, good permeability and appreciable thermo-responsibility. By changing pH of the mobile phase and temperature of the column in turn, the column can separate three model proteins at the mobile phase velocity up to 2528cmh(-1). A separation mechanism of this matrix was also proposed. All results indicate that the dual thermo- and pH-responsive chromatography matrix has great potentials in 'green' high-speed protein chromatography.
已提出使用接枝有聚（N-异丙基丙烯酰胺-共-甲基丙烯酸丁酯-共-N，N-二甲基氨基丙基丙烯酰胺）（P（NIPAM-共-BMA-共-DMAPAAM））刷的超大孔聚苯乙烯微球（GPM）作为基质的双温敏和pH响应色谱法。原子转移自由基聚合（ATRP）引发剂首先通过与2-溴异丁酰溴的傅克酰化反应偶联到GPM上。然后通过表面引发的ATRP将双响应聚合物刷接枝到GPM上。详细表征了基质（GPM-P（NIPAM-共-BMA-共-DMAPAAM））的表面组成、超大孔结构、蛋白质吸附和双响应色谱性能。结果表明，GPM成功地接枝了热响应性阳离子聚合物刷，并且超大孔结构得到了很好的保持。填充有GPM-P（NIPAM-共-BMA-共-DMAPAAM）的色谱柱具有低背压、良好的渗透性和明显的热响应性。通过依次改变流动相的pH值和色谱柱的温度，该色谱柱能够在高达2528cmh（-1）的流动相流速下分离三种模型蛋白质。还提出了该基质的分离机制。所有结果表明，双温敏和pH响应色谱基质在“绿色”高速蛋白质色谱中具有巨大潜力。
