Lee Micky Fu Xiang, Chan Eng Seng, Tam Kam Chiu, Tey Beng Ti
Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 46150 Bandar Sunway, Selangor, Malaysia.
Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 46150 Bandar Sunway, Selangor, Malaysia; Multidisciplinary Platform of Advanced Engineering, Monash University Malaysia, Jalan Lagoon Selatan, 46150 Bandar Sunway, Selangor, Malaysia.
J Chromatogr A. 2015 May 15;1394:71-80. doi: 10.1016/j.chroma.2015.03.034. Epub 2015 Mar 21.
A thermo-responsive random copolymer, POEGMA (poly(oligoethylene glycol) methacrylate) grafted on cationized agarose adsorbent was used for size selective protein adsorption. The effects of OEGMA300 ((oligoethylene glycol) methyl ether methacrylate, Mn=300g/mol) content and temperature on the adsorption of bovine serum albumin (BSA) were evaluated. Increasing the content of OEGMA300 resulted a reduction in BSA adsorption due to the enhanced shielding effect of OEGMA300 chains. Grafting of POEGMA chains onto cationized agarose adsorbent reduced the BSA adsorption by more than 95% at 26.5°C, which is below the LCST (lower critical solution temperature) of POEGMA. The BSA adsorption capacities for adsorbents grafted with 10 and 20mol% of OEGMA300 decreased by 48% and 46% respectively at 38°C, a temperature higher than their LCSTs. The temperature-dependent adsorption of BSA on the adsorbents was attributed to changes in the polymer conformation. The thermal transition of grafted POEGMA conformation exposed the ligand when the temperature was increased. Myoglobin (Myo), which was smaller than BSA, its adsorption behavior was less dependent on the polymer conformation. The adsorption of myoglobin onto the adsorbent with and without POEGMA showed similar percentage of reduction whereas the adsorption of BSA onto the adsorbent with POEGMA decreased by 7.6 times compared to the one without POEGMA. The packed bed of POEGMA grafted adsorbent was used for flow through separation of a protein mixture consisted of virus-like particle, Hepatitis B virus-like particle (HBVLP), BSA and insulin aspart. The recovery of HBVLP in 20mol% of OEGMA300 grafted adsorbent was increased by 19% compared to ungrafted adsorbent. The flow through of BSA can be reduced by increasing the operating temperature above LCST of 20mol% of OEGMA300 while the smaller protein, insulin aspart, remained adsorbed onto the cationized surface. Hence, this thermo-responsive adsorbent has a potential for size-selective separation of protein especially for the recovery of large biomolecule.
一种接枝于阳离子化琼脂糖吸附剂上的热响应性无规共聚物聚（低聚乙二醇）甲基丙烯酸酯（POEGMA）用于尺寸选择性蛋白质吸附。评估了甲基丙烯酸（低聚乙二醇）甲醚（OEGMA300，Mn = 300 g/mol）含量和温度对牛血清白蛋白（BSA）吸附的影响。由于OEGMA300链的屏蔽作用增强，增加OEGMA300的含量导致BSA吸附量降低。在26.5°C（低于POEGMA的低临界溶液温度（LCST））下，将POEGMA链接枝到阳离子化琼脂糖吸附剂上使BSA吸附量降低了95%以上。在38°C（高于其LCST的温度）下，接枝10mol%和20mol% OEGMA300的吸附剂对BSA的吸附容量分别降低了48%和46%。BSA在吸附剂上的温度依赖性吸附归因于聚合物构象的变化。当温度升高时，接枝的POEGMA构象的热转变使配体暴露出来。肌红蛋白（Myo）比BSA小，其吸附行为对聚合物构象的依赖性较小。肌红蛋白在有和没有POEGMA的吸附剂上的吸附减少百分比相似，而与没有POEGMA的吸附剂相比，BSA在有POEGMA的吸附剂上的吸附量降低了7.6倍。接枝POEGMA的吸附剂填充床用于流过分离由病毒样颗粒、乙型肝炎病毒样颗粒（HBVLP）、BSA和门冬胰岛素组成的蛋白质混合物。与未接枝的吸附剂相比，接枝20mol% OEGMA300的吸附剂中HBVLP的回收率提高了19%。通过将操作温度提高到20mol% OEGMA300的LCST以上，可以减少BSA的流过，而较小的蛋白质门冬胰岛素仍吸附在阳离子化表面上。因此，这种热响应性吸附剂具有尺寸选择性分离蛋白质的潜力，特别是对于回收大型生物分子。
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