Multistage continuous countercurrent diafiltration for formulation of monoclonal antibodies
Multistage continuous countercurrent diafiltration for formulation of monoclonal antibodies
复制标题
用于单克隆抗体配制的多级连续逆流渗滤
DOI：
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
A. Zydney
中科院分区：
文献类型：
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作者：
Mario G. Jabra;Christopher J. Yehl;A. Zydney
人们对开发用于生产单克隆抗体产品的完全集成和连续的生物制造工艺越来越感兴趣。最近的一项研究证明了使用两级逆流渗滤 (DF) 工艺进行连续产品配制的可行性，但该系统没有为大多数应用提供足够水平的缓冲液交换。本研究的目的是设计和测试一个三级逆流 DF 系统，该系统可以在连续运行 24 小时内实现至少 99.9% 的缓冲液交换。使用人免疫球蛋白 G 的浓缩溶液作为模型蛋白获得实验数据，并使用维生素 B12 去除的程度来追踪 DF 的程度。具有 Delta 30 kD 再生纤维素膜的 Pall Cadence™ 在线浓缩器用于三个阶段，以在单程中实现高转化率。三级系统运行稳定，维生素 B12 去除率 >99.9%，并且在整个 24 小时内压力增加最小。使用氢氧化钠有效清洁模块，水渗透率几乎完全恢复。提出了一个简单的经济分析，说明了此类逆流分级 DF 工艺的缓冲液使用量和膜成本之间的权衡。研究结果为抗体配制连续工艺的设计和操作提供了重要的见解。
There is growing interest in the development of fully integrated and continuous biomanufacturing processes for the production of monoclonal antibody products. A recent study has demonstrated the feasibility of using a two‐stage countercurrent diafiltration (DF) process for continuous product formulation, but this system did not provide sufficient levels of buffer exchange for most applications. The objective of this study was to design and test a three‐stage countercurrent DF system that could achieve at least 99.9% buffer exchange over 24 hr of continuous operation. Experimental data were obtained using concentrated solutions of human immunoglobulin G as a model protein, with the extent of vitamin B12 removal used to track the extent of DF. Pall Cadence™ inline concentrators with Delta 30 kD regenerated cellulose membranes were used in the three stages to achieve high conversion in a single pass. The three‐stage system showed stable operation with >99.9% vitamin B12 removal and a minimal increase in pressure over the full 24 hr. Modules were effectively cleaned using sodium hydroxide, with nearly complete recovery of water permeability. A simple economic analysis was presented that accounts for the trade‐offs between quantity of buffer used and membrane costs for this type of countercurrent staged DF process. The results provide important insights to the design and operation of a continuous process for antibody formulation.