Steinebach Fabian, Ulmer Nicole, Decker Lara, Aumann Lars, Morbidelli Massimo
Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland.
ChromaCon AG, Zurich, Switzerland.
J Chromatogr A. 2017 Apr 7;1492:19-26. doi: 10.1016/j.chroma.2017.02.049. Epub 2017 Feb 23.
As typical for separation processes, single unit batch chromatography exhibits a trade-off between purity and yield. The twin-column MCSGP (multi-column countercurrent solvent gradient purification) process allows alleviating such trade-offs, particularly in the case of difficult separations. In this work an efficient and reliable procedure for the design of the twin-column MCSGP process is developed. This is based on a single batch chromatogram, which is selected as the design chromatogram. The derived MCSGP operation is not intended to provide optimal performance, but it provides the target product in the selected fraction of the batch chromatogram, but with higher yield. The design procedure is illustrated for the isolation of the main charge isoform of a monoclonal antibody from Protein A eluate with ion-exchange chromatography. The main charge isoform was obtained at a purity and yield larger than 90%. At the same time process related impurities such as HCP and leached Protein A as well as aggregates were at least equally well removed. Additionally, the impact of several design parameters on the process performance in terms of purity, yield, productivity and buffer consumption is discussed. The obtained results can be used for further fine-tuning of the process parameters so as to improve its performance.
作为分离过程的典型情况，单柱间歇色谱法在纯度和产率之间存在权衡。双塔MCSGP（多柱逆流溶剂梯度纯化）工艺能够缓解这种权衡，特别是在难分离的情况下。在这项工作中，开发了一种高效且可靠的双塔MCSGP工艺设计程序。这基于一张单批色谱图，该图被选作设计色谱图。所推导的MCSGP操作并非旨在提供最佳性能，而是在批色谱图的选定馏分中提供目标产物，但产率更高。通过离子交换色谱法从蛋白A洗脱液中分离单克隆抗体的主要电荷异构体，对该设计程序进行了说明。主要电荷异构体的纯度和产率均大于90%。同时，与工艺相关的杂质如宿主细胞蛋白、洗脱的蛋白A以及聚集体也至少能被同样有效地去除。此外，还讨论了几个设计参数对工艺性能在纯度、产率、生产率和缓冲液消耗方面的影响。所获得的结果可用于进一步微调工艺参数，以提高其性能。
