A model-based approach for scaling up chromatographic capture step was developed. The purification of human basic fibroblast growth factor protein 2 (FGF2) from an E. coli homogenate on a cation exchange resin was selected as a case study. Non-ideal effects accompanying the capture operation were examined, including: reduction in the protein diffusivity in the presence of the homogenate, competitive adsorption between FGF2 and undefined impurities, and flow behavior in external column volumes. The viscosity of the homogenate was measured as a function of dilution degree and shear stress, and its contribution to the diffusivity reduction was quantified. A dynamic model was formulated which accounted for underlying kinetic and thermodynamic dependencies. The model parameters were determined for a lab scale system using a small 2-mL column. The model was successfully used to scale up the capture operation from the lab scale column to a preparative bench scale column of about 1 L volume.

中文翻译：

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扩大了澄清细菌匀浆的色谱捕获步骤-传质限制和杂质竞争性吸附的影响。

开发了一种基于模型的放大色谱捕获步骤的方法。选择从阳离子交换树脂上的大肠杆菌匀浆中纯化人碱性成纤维细胞生长因子蛋白2（FGF2）作为案例研究。检查了伴随捕获操作的非理想影响，包括：匀浆存在下蛋白质扩散率降低，FGF2和不确定杂质之间的竞争性吸附以及外部色谱柱体积的流动行为。测量均质物的粘度作为稀释度和剪切应力的函数，并定量其对扩散率降低的贡献。建立了一个动力学模型，该模型考虑了潜在的动力学和热力学依赖性。使用2-mL小型色谱柱为实验室规模的系统确定模型参数。