Preparation and characterization of a thermoresponsive gigaporous medium for high-speed protein chromatography.
Preparation and characterization of a thermoresponsive gigaporous medium for high-speed protein chromatography.
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DOI：
10.1016/j.aca.2014.09.034
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发表时间：
2015
影响因子：
6.2
通讯作者：
Jian-Bo Qu;Yan-Li Chen;Guan-Sheng Huan;Weiqing Zhou;Jian-guo Liu;Hu Zhu;Xiao-yun Zhang
中科院分区：
文献类型：
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作者：
Jian-Bo Qu;Yan-Li Chen;Guan-Sheng Huan;Weiqing Zhou;Jian-guo Liu;Hu Zhu;Xiao-yun Zhang
采用表面引发原子转移自由基聚合（ATRP）技术，将聚（N-异丙基丙烯酰胺-甲基丙烯酸丁酯共聚物）（P（NIPAM-co-BMA））刷接枝到多孔聚苯乙烯（PS）微球上，制备了一种机械强度高、化学稳定性好、对生物大分子传质速率高的高速温敏介质。对制备的介质（PS-P（NIPAM-co-BMA））的巨孔结构、表面化学组成、静态蛋白吸附和温敏色谱性能进行了详细表征。结果表明，PS微球成功地接枝上了P（NIPAM-co-BMA）刷，并保持了稳定的巨孔结构。接枝后，蛋白质在PS微球上的非特异性吸附大大降低。PS-P（NIPAM-co-BMA）填充柱具有较低的背压和显著的热响应性。通过简单地改变柱温，它能够在高达2167 cm h−1的移动的相速度下分离三种模型蛋白质。总之，ATRP法制备的温敏聚合物刷接枝多孔PS微球在“绿色”高速蛋白质制备色谱中具有很好的应用前景。
A high-speed thermoresponsive medium was developed by grafting poly(N-isopropylacrylamide-co-butyl methacrylate) (P(NIPAM-co-BMA)) brushes onto gigaporous polystyrene (PS) microspheres via surface-initiated atom transfer radical polymerization (ATRP) technique, which has strong mechanical strength, good chemical stability and high mass transfer rate for biomacromolecules. The gigaporous structure, surface chemical composition, static protein adsorption, and thermoresponsive chromatographic properties of prepared medium (PS–P(NIPAM-co-BMA)) were characterized in detail. Results showed that the PS microspheres were successfully grafted with P(NIPAM-co-BMA) brushes and that the gigaporous structure was robustly maintained. After grafting, the nonspecific adsorption of proteins on PS microspheres was greatly reduced. A column packed with PS–P(NIPAM-co-BMA) exhibited low backpressure and significant thermo-responsibility. By simply changing the column temperature, it was able to separate three model proteins at the mobile phase velocity up to 2167 cm h−1. In conclusion, the thermoresponsive polymer brushes grafted gigaporous PS microspheres prepared by ATRP are very promising in ‘green’ high-speed preparative protein chromatography.