Thermo- and pH-responsive polymer brushes-grafted gigaporous polystyrene microspheres as a high-speed protein chromatography matrix.
Thermo- and pH-responsive polymer brushes-grafted gigaporous polystyrene microspheres as a high-speed protein chromatography matrix.
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DOI：
10.1016/j.chroma.2016.02.072
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发表时间：
2016-04
期刊：
影响因子：
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通讯作者：
Jian-Bo Qu;Yu-Liang Xu;Jun-Yi Liu;Jingbin Zeng;Yan-Li Chen;Weiqing Zhou;Jian-guo Liu
中科院分区：
文献类型：
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作者：
Jian-Bo Qu;Yu-Liang Xu;Jun-Yi Liu;Jingbin Zeng;Yan-Li Chen;Weiqing Zhou;Jian-guo Liu
以聚(N-异丙基丙烯酰胺-甲基丙烯酸丁酯-N，N-二甲氨基丙基丙烯酰胺)(P(NIPAM-co-BMA-co-DMAPAAM))刷接枝巨孔聚苯乙烯微球(GPM)为基质，建立了温度和pH双重响应层析体系。首次将原子转移自由基聚合(ATRP)引发剂与2-溴异丁基溴通过Friedel-Craft酰化反应偶联到GPM上。然后通过表面引发的ATRP将双响应性聚合物刷接枝到GPM上。对基质(GPM-P(NIPAM-co-BMA-co-DMAPAAM))的表面组成、巨孔结构、蛋白质吸附和双响应层析性能进行了详细的表征。结果表明，热响应型阳离子聚合物刷成功地接枝了GPM，并保持了巨大的孔道结构。GPM-P(NIPAM-co-BMA-co-DMAPAAM)柱具有较低的反压、良好的渗透性和较好的热响应性。通过改变流动相的pH值和柱温，在流动相速度高达2528cmh−1的条件下，可以分离出3种模型蛋白质。这些结果表明，这种温度和pH双重响应的层析基质在绿色高速蛋白质层析中具有很大的潜力。
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 2528 cm h−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.