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Development of multimodal membrane adsorbers for antibody purification using atom transfer radical polymerization

机译:利用原子转移自由基聚合技术开发用于抗体纯化的多峰膜吸附器

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This contribution describes a graft polymerization strategy to prepare multimodal membranes, a new class of high-productivity adsorptive materials for the purification of therapeutic proteins. Surfaceinitiated atom transfer radical polymerization was used to graft poly(glycidyl methacrylate) 'tentacles' from the pore surfaces of macroporous regenerated cellulose membranes. Subsequently, 4-mercaptobenzoic acid was coupled to the membranes by an epoxide ring-opening reaction. ATR-FTIR measurements support successful ligand incorporation. Graft polymerization studies from cellulosecoated silicon substrates were done in parallel to measure the thickness evolution of the polymer coating, which plays an important role on protein binding capacities. Protein binding experiments with bovine immunoglobulin G show that the multimodal membranes have high equilibrium capacities, up to 150 mg IgG/mL. The binding capacities are pH-dependent, with maximum binding at pH near the protein isoelectric point. Characteristic of multimodal adsorbers, the membranes retain about 70% of their equilibrium binding capacity at moderate ionic strength (300 mM) and about 40% at high ionic strength (1.6 M).
机译:这一贡献描述了用于制备多峰膜的接枝聚合策略,多峰膜是用于纯化治疗性蛋白质的新型一类高生产率吸附材料。表面引发的原子转移自由基聚合用于从大孔再生纤维素膜的孔表面接枝聚(甲基丙烯酸缩水甘油酯)“触角”。随后,通过巯基开环反应将4-巯基苯甲酸偶联到膜上。 ATR-FTIR测量支持配体的成功结合。平行进行了纤维素涂层硅基材的接枝聚合研究,以测量聚合物涂层的厚度演变,这对蛋白质结合能力起着重要作用。牛免疫球蛋白G的蛋白质结合实验表明,多峰膜具有很高的平衡能力,最高可达150 mg IgG / mL。结合能力是pH依赖性的,在接近蛋白质等电点的pH下具有最大结合。多峰吸附器的特点是,在中等离子强度(300 mM)时,膜保留约70%的平衡结合能力;在高离子强度(1.6 M)时,约40%。

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