Chemical Engineering Discipline, School of Engineering, Monash University Malaysia, Selangor, Malaysia.
Faculty of Chemical and Natural Resources Engineering, Universiti Malaysia Pahang, Pahang, Malaysia.
J Sep Sci. 2018 May;41(10):2119-2129. doi: 10.1002/jssc.201700823. Epub 2018 Mar 15.
Hepatitis B virus-like particles expressed in Escherichia coli were purified using anion exchange adsorbents grafted with polymer poly(oligo(ethylene glycol) methacrylate) in flow-through chromatography mode. The virus-like particles were selectively excluded, while the relatively smaller sized host cell proteins were absorbed. The exclusion of virus-like particles was governed by the accessibility of binding sites (the size of adsorbents and the charge of grafted dextran chains) as well as the architecture (branch-chain length) of the grafted polymer. The branch-chain length of grafted polymer was altered by changing the type of monomers used. The larger adsorbent (90 μm) had an approximately twofold increase in the flow-through recovery, as compared to the smaller adsorbent (30 μm). Generally, polymer-grafted adsorbents improved the exclusion of the virus-like particles. Overall, the middle branch-chain length polymer grafted on larger adsorbent showed optimal performance at 92% flow-through recovery with a purification factor of 1.53. A comparative study between the adsorbent with dextran grafts and the polymer-grafted adsorbent showed that a better exclusion of virus-like particles was achieved with the absorbent grafted with inert polymer. The grafted polymer was also shown to reduce strong interaction between binding sites and virus-like particles, which preserved the particles' structure.
在大肠杆菌中表达的乙型肝炎病毒样颗粒采用在流穿模式下用接枝有聚合物聚（聚（乙二醇）甲基丙烯酸酯）的阴离子交换吸附剂进行纯化。病毒样颗粒被选择性排斥，而相对较小的宿主细胞蛋白被吸附。病毒样颗粒的排除受结合位点的可及性（吸附剂的大小和接枝葡聚糖链的电荷）以及接枝聚合物的结构（支链长度）控制。通过改变使用的单体类型来改变接枝聚合物的支链长度。与较小的吸附剂（30 μm）相比，较大的吸附剂（90 μm）的流穿回收率增加了约两倍。通常，接枝聚合物的吸附剂可提高病毒样颗粒的排除率。总体而言，在 92%的流穿回收率和 1.53 的纯化因子下，接枝在较大吸附剂上的中等支链长度聚合物表现出最佳性能。接枝有葡聚糖的吸附剂和接枝聚合物的吸附剂之间的比较研究表明，用惰性聚合物接枝的吸附剂可实现更好的病毒样颗粒排除。接枝聚合物还可减少结合位点与病毒样颗粒之间的强相互作用，从而保持颗粒的结构。
