The National Creative Research Initiative Center for Intelligent Hybrids, School of Chemical and Biological Engineering , Seoul National University , Seoul 08826 , Korea.
Biomacromolecules. 2018 Mar 12;19(3):951-961. doi: 10.1021/acs.biomac.7b01736. Epub 2018 Feb 15.
Reactive pentafluorophenyl acrylate (PFPA) polymer brushes grafted on silica particles were prepared using surface-initiated reversible addition and fragmentation chain transfer polymerization. The polymer brush was successfully immobilized with antibody, then used for protein separation. The immunoprecipitated proteins showed successful enrichment of target protein, with reduced nonspecific background and less contamination from eluted antibodies. To further improve protein recovery, the hydrophobic poly(PFPA) brush was modified with hydrophilic poly(ethylene glycol) (PEG). The partially PEG-substituted poly(PFPA) brush showed better dispersion in aqueous solution, leading to improved antibody immobilization efficiency. By optimizing both the brush molecular weight and the degree of PEG substitution, an optimal balance between surface hydrophilicity and number of available PFP units was found, leading to efficient target protein purification. This study shows that poly(PFPA) platform offers a versatile approach to prepare biomolecule-activated surfaces with tunable surface property, which has potential applications in protein separation and other areas.
采用表面引发可逆加成-断裂链转移聚合，制备了接枝在硅胶粒子上的活性五氟苯基丙烯酸酯（PFPA）聚合物刷。成功地将聚合物刷固定化抗体，然后用于蛋白质分离。免疫沉淀的蛋白质显示出目标蛋白的成功富集，非特异性背景减少，洗脱抗体的污染减少。为了进一步提高蛋白质回收率，用亲水性聚乙二醇（PEG）对疏水性聚（PFPA）刷进行改性。部分 PEG 取代的聚（PFPA）刷在水溶液中表现出更好的分散性，从而提高了抗体的固定化效率。通过优化刷的分子量和 PEG 取代度，找到了表面亲水性和可用 PFP 单元数量之间的最佳平衡，从而实现了目标蛋白质的高效纯化。本研究表明，聚（PFPA）平台提供了一种通用的方法来制备具有可调表面性能的生物分子激活表面，在蛋白质分离和其他领域具有潜在的应用。
