Biothermodynamics, TUM School of Life Sciences, Technical University of Munich, Freising, Germany; Corporate Research, Sartorius Stedim Biotech GmbH, Göttingen, Germany.
Biothermodynamics, TUM School of Life Sciences, Technical University of Munich, Freising, Germany.
J Chromatogr A. 2024 Sep 13;1732:465201. doi: 10.1016/j.chroma.2024.465201. Epub 2024 Jul 21.
Protein A affinity membrane adsorbers are a promising alternative to resins to intensify the manufacturing of monoclonal antibodies. This study examined the process performance of convective diffusive membrane adsorbers operated in batch and continuous multi-column mode. Therefore, three different processes were compared regarding membrane utilization, productivity, and buffer consumption: the batch process, the rapid cycling parallel multi-column chromatography process, and the rapid cycling simulated moving bed process. The influence of the monoclonal antibody loading concentration (between 0.5 g L and 5.2 g L) and the loading flow rate (between 1.25 MV min and 10 MV min) on the monoclonal antibody binding behavior of the membrane adsorber were studied with breakthrough curve experiments. The determined breakthrough curves were used to calculate the monoclonal antibody dynamic binding capacity, the duration of the loading steps for each process, and the number of required membrane adsorbers for the continuous processes rapid cycling parallel multi-column chromatography and rapid cycling simulated moving bed. The highest productivity for the batch (176 g L h) and rapid cycling parallel multi-column chromatography process (176 g L h) was calculated for high monoclonal antibody loading concentrations and low loading flow rates. In contrast, the rapid cycling simulated moving bed process achieved the highest productivity (217 g L h) for high monoclonal antibody loading concentrations and loading flow rates. Furthermore, due to the higher membrane utilization, the buffer consumption of the rapid cycling simulated moving bed process (1.1 L g) was up to 1.9 times lower than that of the batch or rapid cycling parallel multi-column chromatography operation (2.1 L g).
Protein A 亲和膜吸附剂是强化单克隆抗体生产的一种有前途的树脂替代物。本研究考察了在间歇和连续多柱模式下操作的对流扩散膜吸附器的过程性能。因此，就膜利用、生产率和缓冲液消耗而言，比较了三种不同的工艺：间歇工艺、快速循环并行多柱色谱工艺和快速循环模拟移动床工艺。通过穿透曲线实验研究了单克隆抗体装载浓度（0.5 克/升至 5.2 克/升之间）和装载流速（1.25MVmin 至 10MVmin 之间）对膜吸附器中单克隆抗体结合行为的影响。所确定的穿透曲线用于计算单克隆抗体的动态结合容量、每个过程的装载步骤持续时间以及连续过程快速循环并行多柱色谱和快速循环模拟移动床所需的膜吸附器数量。对于高单克隆抗体装载浓度和低装载流速，间歇（176g/Lh）和快速循环并行多柱色谱工艺（176g/Lh）的生产率最高。相比之下，对于高单克隆抗体装载浓度和装载流速，快速循环模拟移动床工艺的生产率最高（217g/Lh）。此外，由于膜利用率较高，快速循环模拟移动床工艺的缓冲液消耗（1.1L/g）比间歇或快速循环并行多柱色谱操作（2.1L/g）低 1.9 倍。
