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Continuous Production of Proteins: Integration of Polishing Using MCSGP

机译：连续生产蛋白质：使用MCSGP进行抛光整合

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摘要

Continuous counter-current processes for polishing steps overcome the purity-yield tradeoff of classical batch chromatography by internal recycling of overlapping regions of the chromatogram. Particularly in the case of challenging separation tasks this leads for the same product □uality to significantly higher yields and productivity. Processes using these fundamental principals have been successfully implemented in the chemical industry up to multi-ton production. In an integrated continuous process, with a perfusion bioreactor and a continuous capture process, the product □uality of the capture product is different compared to classical batch processing. The lower residence time in the bioreactor and the higher loading in multi-column capture steps lead to polishing feed streams with higher concentration and less impurities. A cascade of beneficial correlations results in higher titer. Hence the polishing process parameters need to be adjusted accordingly. In this work the process design as well as the results for monoclonal antibody polishing with a twin-column MCSGP setup is presented, where the feed stream was obtained from an integrated continuous cultivation and capture process. The process was designed accordingly and the resulting product □uality in terms of aggregate and fragment content, as well as charge isoforms is compared to classical batch chromatography. The internal recycling of the product resulted in a higher productivity and yield for the multicolumn setup.
机译：抛光步骤的连续逆流工艺通过色谱图重叠区域的内部循环克服了传统分批色谱法的纯度-收率折衷。特别是在具有挑战性的分离任务中，这将导致相同的产品质量，从而显着提高产量和生产率。使用这些基本原理的过程已成功地在化学工业中实施，直至多吨生产。在集成的连续过程中，通过灌注生物反应器和连续捕获过程，捕获产物的产品质量与传统的分批处理相比有所不同。在生物反应器中的停留时间越短，在多柱捕获步骤中的装载量越高，就可以精制浓度更高且杂质更少的进料流。一系列有益的相关性导致更高的效价。因此，抛光工艺参数需要相应地调整。在这项工作中，介绍了工艺设计以及使用双柱MCSGP设置进行单克隆抗体抛光的结果，其中进料流是从集成的连续培养和捕获过程中获得的。相应地设计了该方法，并将所得产物在聚集体和碎片含量以及电荷同工型方面的质量与经典的批量色谱法进行了比较。产品的内部回收利用为多柱设置带来了更高的生产率和良率。

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