Schwellenbach Jan, Zobel Steffen, Taft Florian, Villain Louis, Strube Jochen
Sartorius Stedim Biotech GmbH, Göttingen 37079, Germany.
Institute for Separation and Process Technology, Clausthal University of Technology, Clausthal-Zellerfeld 38678, Germany.
Bioengineering (Basel). 2016 Oct 2;3(4):24. doi: 10.3390/bioengineering3040024.
Monoclonal antibodies (mAb) currently dominate the market for protein therapeutics. Because chromatography unit operations are critical for the purification of therapeutic proteins, the process integration of novel chromatographic stationary phases, driven by the demand for more economic process schemes, is a field of ongoing research. Within this study it was demonstrated that the description and prediction of mAb purification on a novel fiber based cation-exchange stationary phase can be achieved using a physico-chemical model. All relevant mass-transport phenomena during a bind and elute chromatographic cycle, namely convection, axial dispersion, boundary layer mass-transfer, and the salt dependent binding behavior in the fiber bed were described. This work highlights the combination of model adaption, simulation, and experimental parameter determination through separate measurements, correlations, or geometric considerations, independent from the chromatographic cycle. The salt dependent binding behavior of a purified mAb was determined by the measurement of adsorption isotherms using batch adsorption experiments. Utilizing a combination of size exclusion and protein A chromatography as analytic techniques, this approach can be extended to a cell culture broth, describing the salt dependent binding behavior of multiple components. Model testing and validation was performed with experimental bind and elute cycles using purified mAb as well as a clarified cell culture broth. A comparison between model calculations and experimental data showed a good agreement. The influence of the model parameters is discussed in detail.
单克隆抗体（mAb）目前在蛋白质治疗药物市场中占据主导地位。由于色谱单元操作对于治疗性蛋白质的纯化至关重要，在更经济的工艺方案需求推动下，新型色谱固定相的工艺集成是一个正在进行研究的领域。在本研究中，证明了使用物理化学模型可以实现对新型纤维基阳离子交换固定相上mAb纯化的描述和预测。描述了在结合和洗脱色谱循环过程中所有相关的传质现象，即对流、轴向扩散、边界层传质以及纤维床中盐依赖性结合行为。这项工作强调了通过单独测量、关联或几何考虑来进行模型适配、模拟和实验参数确定的结合，而与色谱循环无关。通过批量吸附实验测量吸附等温线来确定纯化mAb的盐依赖性结合行为。利用尺寸排阻色谱和蛋白A色谱相结合作为分析技术，这种方法可以扩展到细胞培养液，描述多种成分的盐依赖性结合行为。使用纯化的mAb以及澄清的细胞培养液进行实验结合和洗脱循环，进行模型测试和验证。模型计算与实验数据之间的比较显示出良好的一致性。详细讨论了模型参数的影响。
