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The effect of feed quality due to clarification strategy on the design and performance of protein A periodic counter-current chromatography

机译：由于澄清策略对蛋白质的设计和性能澄清策略的影响，周期性逆流色谱

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摘要

The impact of two different quality feeds, derived using two different harvest clarification processes, on protein A periodic counter-current chromatography (PCC) design and performance is investigated. Data from batch experiments were input into a model to design optimal PCC operating parameters specific to each feed material. The two clarification methods were: depth filtration using a wetlaid matrix which has Q-functionality; and a combination of depth filtration and chromatographic clarification, using a Q-functional nonwoven with a high anion exchange capacity (Emphaze (TM) AEX Hybrid Purifier) in which key impurities such as host cell DNA (HCDNA) and host cell proteins (HCP) are removed. The model predicted 34% better productivity for the chromatographically clarified cell culture fluid (CCCF) using a 4 column system, and productivity gains of 28% using only 3 columns enabling the option to simplify the protein A PCC strategy. Experimental validation of the predicted optimized PCC operating parameters using industrially relevant monoclonal antibody (mAb) CCCF feedstock over 100 cycles showed productivity gains of 49% for the chromatographically clarified material. HCP concentration was 11-fold lower, and HCDNA concentration was reduced by 4.4 Log Reduction Value (LRV) in the protein A PCC eluates. This work, therefore, demonstrates that the removal of HCDNA and HCP during clarification is an effective strategy for improving protein A PCC performance. This was achieved using the Emphaze (TM) AEX Hybrid Purifier which can be easily incorporated into a batch or continuous process, in a scalable fashion, without adding additional separate unit operations. (c) 2018 American Institute of Chemical Engineers Biotechnol.
机译：研究了使用两种不同的收获澄清过程的两种不同质量饲料的影响，对蛋白质是周期性反电流色谱（PCC）设计和性能。来自批量实验的数据被输入到模型中以设计特定于每个馈材料的最佳PCC操作参数。两种澄清方法是：使用具有Q型Q型的湿覆基质的深度过滤;和深度过滤和色谱释放的组合，使用具有高阴离子交换能力的Q型功能性非织造物（TM）AEX杂交净化器），其中诸如宿主细胞DNA（HCDNA）和宿主细胞蛋白（HCP）之类的关键杂质被删除了。该模型使用4柱系统预测了色谱柱澄清的细胞培养液（CCCF）的生产率更好的生产率，并且仅使用3柱的生产率增益使得能够简化蛋白质A PCC策略的选项。使用工业相关的单克隆抗体（MAB）CCCF原料的预测优化的PCC操作参数的实验验证超过100个循环的生产率增益显示为色谱柱澄清的材料的49％。 HCP浓度下降11倍，HCDNA浓度降低了4.4蛋白A PCC洗脱液中的降低值（LRV）。因此，这项工作表明，在澄清过程中除去HCDNA和HCP是改善蛋白质PCC性能的有效策略。这是使用的，使用膨胀（TM）AEX混合净化器以可伸缩的方式容易地结合到批量或连续过程中，而无需添加额外的单独单元操作。 （c）2018美国化学工程师学院Biotechnol。

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