新型聚合物单分散温敏色谱固定相的制备及其在生物大分子分离中的应用  

Preparation of New Polymeric Monodisperse Thermosensitive Chromatographic Stationary Phase and Its Application for Separation of Biopolymers

作　　者：欧阳康龙[1] 孙彦璞[1] 龚波林[1] 

机构地区：[1]宁夏大学能源化工重点实验室,银川750021

出　　处：《分析化学》2010年第6期851-854,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.20765004)资助

摘　　要：以2.5μm的单分散聚苯乙烯为种子,乙二醇二甲基丙烯酸酯(EDMA)为交联剂,甲苯和环己醇为致孔剂,采用"一步种子溶胀聚合法"制备了单分散微球;再以过硫酸钾为引发剂将水溶性温敏单体N-异丙基丙烯酰胺(NIPAM)分子引发聚合到微球表面,制备了粒径为7.0μm、分散系数为0.02的单分散交联温敏色谱填料,温敏单体NIPAM的接枝率为5.2%。考察了该填料对标准蛋白质的分离性能、温敏性能、稳定性和重现性以及动态吸附容量对蛋白保留的影响。实验结果表明,该色谱填料对蛋白的分离性能、温敏性能、稳定性及重现性良好,且对溶菌酶的动态吸附容量为32.3mg/g。在疏水模式下,该填料不但可以同时基线分离5种标准蛋白,而且通过改变温度可以有效地将3种在低温下保留时间重叠的蛋白(细胞色素-C、β-乳球蛋白和核糖核酸酶)完全分离。By using 2.5 μm monodisperse polystyrene as seeds, ethylene glycol dimethyl acrylate（EDMA） as crosslinking agent, toluene and cyclohexanol as porogen, one-step of seed swelling polymerization method was applied for the preparation of monodisperse beads. Then the monomer of N-isopropylacrylamide（NTPAM） was polymerized on the surface of monodisperse beads by water-soluble initiator（potassium persulfate） to prepare a 7.0-μm monodisperse thermosensitive chromatographic stationary phase. Its dispersion coefficient is 0.02 and the grafting yield of NIPAM is 5.2%. The stationary phase was evaluated in detail to by determining its separability, thermosensitivity, stability, reproducibility and the effect of the dynamic protein loading capacity on the retention of proteins. The results show that the packing has good separability, thermosensitivity, stability, reproducibility and the dynamic protein loading capacity for lysyme is 32.3mg/g. In the hydrophobic interaction chromatographic model, five standard proteins can be separated by the stationary phase, and cytochome C, β-lactoglobulin, ribonmclease A can be effectively separated by changing the temperature.

关 键 词：N-异丙基丙烯酰胺 单分散温敏色谱固定相 蛋白质分离 

分 类 号：O657.7[理学—分析化学]