Schmitz Fabian, Saballus Martin, Kruse Thomas, Minceva Mirjana, Kampmann Markus
Biothermodynamics, TUM School of Life Sciences, Technical University of Munich, Freising, Germany.
Corporate Research, Sartorius Stedim Biotech GmbH, Göttingen, Germany.
Biotechnol Bioeng. 2025 Feb;122(2):382-394. doi: 10.1002/bit.28884. Epub 2024 Nov 18.
Harmonizing unit operations in the downstream process of monoclonal antibodies (mAbs) has a high potential to overcome throughput limitations and reduce manufacturing costs. This study proposes a streamlined clarification and capture (S-CC) process concept for the continuous processing of cell broth harvested from a connected bioreactor. The process was realized with a fluidized bed centrifuge connected to depth and sterile filters, a surge tank, and a multi-column chromatography (MCC) unit. The MCC unit was operated in the rapid cycling simulated moving bed (RC-BioSMB) mode with five convective diffusive membrane adsorbers (MAs). A control strategy and the surge tank were used to adjust the loading flow rate of the MCC unit. The mAb was recovered with a total process yield of 90%, with high removal of the process-related impurities HCP (2.1 LRV) and DNA (2.9 LRV). Moreover, the S-CC process productivity of 4.2 g h was up to 5.3 times higher than for comparable, hypothetical batch MA processes. In addition, the buffer consumption of the capture step could be reduced from 2.0 L g in batch mode to 1.2 L g in the RC-BioSMB mode. These results demonstrate the high potential of streamlined interconnected unit operations to improve the overall mAb downstream process performance.
协调单克隆抗体（mAb）下游工艺中的单元操作，在克服通量限制和降低制造成本方面具有很大潜力。本研究提出了一种简化的澄清和捕获（S-CC）工艺概念，用于连续处理从连接的生物反应器收获的细胞培养液。该工艺通过连接深度过滤器和无菌过滤器、缓冲罐以及多柱色谱（MCC）单元的流化床离心机实现。MCC单元采用五个对流扩散膜吸附器（MA）以快速循环模拟移动床（RC-BioSMB）模式运行。使用一种控制策略和缓冲罐来调整MCC单元的进料流速。单克隆抗体的总回收率为90%，对工艺相关杂质宿主细胞蛋白（HCP，去除率2.1 LRV）和DNA（去除率2.9 LRV）的去除效果良好。此外，S-CC工艺的生产率为4.2 g/h，比类似的假设分批MA工艺高出5.3倍。此外，捕获步骤的缓冲液消耗可从分批模式下的2.0 L/g降至RC-BioSMB模式下的1.2 L/g。这些结果表明，简化的互连单元操作在提高单克隆抗体下游工艺整体性能方面具有很大潜力。
