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首页> 外文期刊>Biotechnology Progress >Demonstration of the Use of Windows of Operation to Visualize the Effects of Fouling on the Performance of a Chromatographic Step
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Demonstration of the Use of Windows of Operation to Visualize the Effects of Fouling on the Performance of a Chromatographic Step

机译：演示使用操作窗口可视化结垢对色谱步骤性能的影响

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摘要

The high resolution afforded by packed bed chromatography makes it an indispensable operation in the downstream processing of therapeutic molecules. Packed bed performance is however inherently susceptible to changes in feed stream characteristics and fouling processes. The impact of fouling is seldom considered during the early stages of bioprocess development which is concerned with the selection of purification conditions. Instead these are performed with rigorously clarified feeds. Under such conditions, chromatography is effectively treated as an isolated step, independent from its preceding unit operations. In this study, we demonstrate how windows of operation could be used to visualize the impact of changes in the preceding clarification step on the fouling response of a subsequent cation exchange capture step. Laboratory columns (2,5 and 12 cm height) were subjected to varying fouling challenges of Escherichia coli lysate containing different amounts of solids carried over from the previous step. Changes in trans-column pressure drop and breakthrough of the target protein (Fab') were monitored. The limits of operability of the resin were determined with respect to the process material's properties. This information was used to extract the parameters for the adsorption kinetics used in the general rate (GR) model to create windows of operation for manufacturing scale operation.
机译：填充床色谱提供的高分辨率使其成为治疗分子下游加工中必不可少的操作。然而，填充床性能固有地易受进料流特性和结垢过程的变化的影响。在涉及纯化条件选择的生物工艺开发的早期阶段，很少考虑结垢的影响。而是使用严格澄清的提要来执行这些操作。在这样的条件下，色谱法被有效地视为一个分离的步骤，而与之前的单元操作无关。在这项研究中，我们演示了如何使用操作窗口来可视化前面的澄清步骤中的变化对后续阳离子交换捕获步骤的结垢响应的影响。实验室色谱柱（2,5和12 cm高）受到不同程度的结垢挑战，其中大肠杆菌裂解物含有从上一步骤得到的不同量的固体。监测跨柱压降的变化和靶蛋白（Fab'）的穿透。树脂的可操作性极限是根据加工材料的性质确定的。此信息用于提取一般速率（GR）模型中使用的吸附动力学参数，以创建制造规模操作的操作窗口。

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