Osuofa Joshua, Husson Scott M
Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC 29634, USA.
Membranes (Basel). 2023 May 12;13(5):511. doi: 10.3390/membranes13050511.
Protein A chromatography is ubiquitous to antibody purification. The high specificity of Protein A for binding the Fc-region of antibodies and related products enables unmatched clearance of process impurities like host cell proteins, DNA, and virus particles. A recent development is the commercialization of research-scale Protein A membrane chromatography products that can perform capture step purification with short residence times (RT) on the order of seconds. This study investigates process-relevant performance and physical properties of four Protein A membranes: Purilogics Purexa™ PrA, Gore Protein Capture Device, Cytiva HiTrap™ Fibro PrismA, and Sartorius Sartobind Protein A. Performance metrics include dynamic binding capacity, equilibrium binding capacity, regeneration-reuse, impurity clearance, and elution volumes. Physical properties include permeability, pore diameter, specific surface area, and dead volume. Key results indicate that all membranes except the Gore Protein Capture Device operate with flow rate-independent binding capacities; the Purilogics Purexa™ PrA and Cytiva HiTrap Fibro™ PrismA have binding capacities on par with resins, with orders of magnitude faster throughput; and dead volume and hydrodynamics play major roles in elution behavior. Results from this study will enable bioprocess scientists to understand the ways that Protein A membranes can fit into their antibody process development strategies.
蛋白A层析在抗体纯化中无处不在。蛋白A与抗体及相关产物的Fc区域结合具有高度特异性，能够无与伦比地清除宿主细胞蛋白、DNA和病毒颗粒等工艺杂质。最近的一项进展是研究规模的蛋白A膜层析产品商业化，这些产品能够在短至几秒的停留时间内进行捕获步骤纯化。本研究调查了四种蛋白A膜的工艺相关性能和物理性质：Purilogics Purexa™ PrA、戈尔蛋白捕获装置、赛多利斯HiTrap™ Fibro PrismA和 Sartorius Sartobind Protein A。性能指标包括动态结合容量、平衡结合容量、再生再利用、杂质清除和洗脱体积。物理性质包括渗透率、孔径、比表面积和死体积。关键结果表明，除戈尔蛋白捕获装置外，所有膜的结合容量均与流速无关；Purilogics Purexa™ PrA和赛多利斯HiTrap Fibro™ PrismA的结合容量与树脂相当，通量快几个数量级；死体积和流体动力学在洗脱行为中起主要作用。本研究结果将使生物工艺科学家了解蛋白A膜可融入其抗体工艺开发策略的方式。
