Bioprocess Development, Sanofi, 31 New York Avenue, Framingham, MA 01701, USA.
Biotechnol J. 2019 Feb;14(2):e1700733. doi: 10.1002/biot.201700733. Epub 2018 Jun 25.
In this study, the authors compared the impacts of fed-batch and perfusion platforms on process and product attributes for IgG1- and IgG4-producing cell lines. A "plug-and-play" approach is applied to both platforms at bench scale, using commercially available basal and feed media, a standard feed strategy for fed-batch and ATF filtration for perfusion. Product concentration in fed-batch is 2.5 times greater than perfusion, while average productivity in perfusion is 7.5 times greater than fed-batch. PCA reveals more variability in the cell environment and metabolism during the fed-batch run. LDH measurements show that exposure of product to cell lysate is 7-10 times greater in fed-batch. Product analysis shows larger abundances of neutral species in perfusion, likely due to decreased bioreactor residence times and extracellular exposure. The IgG1 perfusion product also has higher purity and lower half-antibody. Glycosylation is similar across both culture modes. The first perfusion harvest slice for both product types shows different glycosylation than subsequent harvests, suggesting that product quality lags behind metabolism. In conclusion, process and product data indicate that intra-lot heterogeneity is decreased in perfusion cultures. Additional data and discussion is required to understand the developmental, clinical and commercial implications, and in what situations increased uniformity would be beneficial.
在这项研究中，作者比较了分批补料和灌注平台对 IgG1 和 IgG4 产生细胞系的工艺和产品属性的影响。在中试规模上，采用“即插即用”的方法，应用市售的基础培养基和补料培养基、分批补料的标准补料策略以及灌注的 ATF 过滤。与灌注相比，分批补料的产物浓度高 2.5 倍，而灌注的平均生产力比分批补料高 7.5 倍。主成分分析（PCA）显示，在分批补料运行过程中，细胞环境和代谢的变化更大。LDH 测量结果表明，产物与细胞裂解物的接触在分批补料中要高 7-10 倍。产物分析显示，在灌注中，中性物质的丰度更高，这可能是由于生物反应器停留时间和细胞外暴露减少所致。IgG1 灌注产物的纯度更高，半抗体含量更低。两种培养模式的糖基化相似。两种产物类型的首次灌注收获切片的糖基化与后续收获的不同，表明产物质量落后于代谢。总之，工艺和产品数据表明，灌注培养中的批次间异质性降低。需要更多的数据和讨论来了解其在开发、临床和商业方面的影响，以及在什么情况下增加均一性会更有利。
