Characterisation of grafted weak anion-exchange methacrylate monoliths
Characterisation of grafted weak anion-exchange methacrylate monoliths
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DOI：
10.1016/j.chroma.2008.08.027
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发表时间：
2008-10-17
影响因子：
4.1
通讯作者：
Strancar, Ales
中科院分区：
文献类型：
--
作者：
Frankovic, Vida;Podgornik, Ales;Strancar, Ales
用甲基丙烯酸缩水甘油酯接枝甲基丙烯酸酯单体，然后用二乙胺修饰环氧基团，制得弱离子交换接枝甲基丙烯酸酯单体。通过测量渗透率确定了接枝层的厚度，发现接枝层的厚度约为90 nm。考察了不同缓冲溶液对压降的影响，指出了pH对接枝整体柱渗透性的影响。蛋白质的分离和结合能力(BC)在280 cm/h的线速度下不受流动的影响。用P-乳球蛋白、牛血清白蛋白(BSA)、甲状腺球蛋白和质粒DNA(PDNA)比较了未接枝和接枝的整体柱的BC。结果发现，在所有情况下，接枝的整体柱的容量都比未接枝的整体柱高2-3.5倍，分别达到105、80、71和17毫克/毫升。结果表明，在负载缓冲液中使用0.1M的氯化钠时，PDNA的容量最大。回收率是可比的，没有检测到超螺旋PDNA形式的降解。对于未接枝和接枝的整体柱，蛋白质z因子相等，这表明尽管接枝的整体柱在较高的离子强度下发生洗脱，但有相同数量的结合位点可用。接枝的整体柱的效率低于未接枝的整体柱。然而，PDNA与RNA和其他杂质的基线分离是从真实样品中实现的。(C)2008年爱思唯尔。版权所有。
A weak ion-exchange grafted methacrylate monolith was prepared by grafting a methacrylate monolith with glycidyl methacrylate and subsequently modifying the epoxy groups with diethylamine. The thickness of the grafted layer was determined by measuring permeability and found to be approximately 90 nm. The effects of different buffer solutions on the pressure drop were examined and indicated the influence of pH on the permeability of the grafted monolith. Protein separation and binding capacity (BC) were found to be flow-unaffected up to a linear velocity of 280 cm/h. A comparison of the BC for the non-grafted and grafted monolith was performed using P-lactoglobulin, bovine serum albumin (BSA), thyroglobulin, and plasmid DNA (pDNA). It was found that the grafted monolith exhibited 2- to 3.5-fold higher capacities (as compared to non-grafted monoliths) in all cases reaching values of 105, 80, 71, and 17 mg/ml, respectively. It was determined that the maximum pDNA capacity was reached using 0.1 M NaCl in the loading buffer. Recovery was comparable and no degradation of the supercoiled pDNA form was detected. Protein z-factors were equal for the non-grafted and grafted monolith indicating that the same number of binding sites are available although elution from the grafted monolith occurred at higher ionic strengths. The grafted monolith exhibited lower efficiency than the non-grafted ones. However, the baseline separation of pDNA from RNA and other impurities was achieved from a real sample. (C) 2008 Elsevier BY. All rights reserved.