Preparation and characterization of polyethyleneimine modified ion-exchanger based on poly(methacrylate-co-ethylene dimethacrylate) monolith.
Preparation and characterization of polyethyleneimine modified ion-exchanger based on poly(methacrylate-co-ethylene dimethacrylate) monolith.
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DOI：
10.1016/j.chroma.2007.02.032
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发表时间：
2007-04
期刊：
影响因子：
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通讯作者：
Manyi Wang;Jun Xu;Xin Zhou;T. Tan
中科院分区：
文献类型：
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作者：
Manyi Wang;Jun Xu;Xin Zhou;T. Tan
以100 mm ×4.6mm内径的聚甲基丙烯酸酯-二甲基丙烯酸乙二酯共聚物（PMMA-co-ethylenedimethacrylate，PEI）为基体材料，制备了PEI改性的离子交换剂。不锈钢管，庚烷作为致孔溶剂，在65°C下12小时。大于500 nm的孔占PEI整体柱总孔体积的85%，为分离提供了更好的渗透性。随着PEI分子量的增加，柱上牛血清白蛋白（BSA）的结合能力增强，表明刷状配体从柱表面发散，通过多个结合位点捕获更多的蛋白质。滴定实验和BSA保留率随移动的pH的变化表明，该整体柱具有较弱的离子交换性能，当移动的相中NaCl含量大于0.5M时，BSA在整体柱上的回收率可达97%.前沿分析和BSA梯度洗脱表明，PEI整体柱在色谱过程中提供了快速的传质，使动态结合容量和柱效在高流速下保持恒定。在1445 cm/h的流速下，在3 min内实现了溶菌酶、血红蛋白和BSA三种模式蛋白混合物的快速分离。这表明PEI整体柱用于蛋白质的快速分析和分离的潜力。
A polyethyleneimine (PEI) modified ion-exchanger was prepared based on poly(methacrylate-co-ethylene dimethacrylate) monolith cast in 100mm×4.6mm I.D. stainless steel tube with heptane as the porogenic solvent at 65°C for 12h. The pores larger than 500nm presented 85% of total pore volume of PEI monolith and provided the better permeability for separation. Bovine serum albumin (BSA) binding capacity on the column was enhanced with increasing the molecular weight of PEI, indicated that the brush ligand emanated from the surface and captured more protein by multiple binding sites. Titration experiment as well as BSA retention versus the pH of mobile phase showed that the monolith exhibited weak ion-exchange property, and recovered BSA on the monolith reached 97% when NaCl content in mobile phase was higher than 0.5M. Frontal analysis and gradient elution of BSA indicated that PEI monolith provided the rapid mass transfer in chromatographic procedure, which made the dynamic binding capacities as well as column efficiency keep as constants at high operating flow rate. Fast separation of three mode proteins mixture (lysozyme, hemoglobin and BSA) on the monolith was achieved within 3min at velocity of 1445cm/h. This demonstrated the potential of PEI monolith for the rapid analysis and separation of proteins.