Balassi Vincent, Otto Mary, Kretzmer Corey, Petersen Amber, McLaurin Channing, Mahadevan Jana, Gustin Jason, Borgschulte Trissa, Razafsky David
Expression Systems and Novel Biopharmaceutical Materials, MilliporeSigma, Saint Louis, Missouri, USA.
Cell Culture Media and Process Development, MilliporeSigma, Saint Louis, Missouri, USA.
Biotechnol Prog. 2025 May-Jun;41(3):e70003. doi: 10.1002/btpr.70003. Epub 2025 Feb 19.
As the industry continues to explore the benefits of continuous and intensified manufacturing, it is important to assure that the cell line development (CLD) workflows in practice today are well suited to generate clones that meet the unique challenges associated with these processes. Most cell lines used in intensified processes are currently developed using traditional fed-batch CLD workflows followed by adaptation of these cell lines to perfusion processes. This method maybe suboptimal as fed-batch CLD workflows select clones which produce high volumetric titers irrespective of cell growth rate and specific productivity (qP). Although sufficient for fed-batch processes, performance of cells derived from this traditional CLD workflow may not be maintained in perfusion processes, where an intricate balance of performance parameters is needed. Until now, a thorough investigation into the effect of the CLD workflow on top clone performance in perfusion processes has not been conducted. Here, we show how the CLD workflow impacts cell performance in both fed-batch and perfusion processes, emphasizing the advantages of adopting a perfusion-specific CLD workflow which includes the use of medium specially designed for expansion and production in a perfusion setting, scale-down models which more accurately simulate perfusion process, and the adoption of perfusion-specific cell line selection criteria. Together, this results in the development of more efficient cell lines, fit for continuous and intensified processing.
随着该行业不断探索连续强化生产的益处,确保当今实际应用中的细胞系开发(CLD)工作流程适合生成能够应对这些工艺所带来独特挑战的克隆体非常重要。目前,强化工艺中使用的大多数细胞系是通过传统的补料分批CLD工作流程开发的,随后将这些细胞系适应灌注工艺。这种方法可能并非最佳选择,因为补料分批CLD工作流程选择的克隆体是那些产生高体积滴度的克隆体,而不考虑细胞生长速率和比生产率(qP)。虽然对于补料分批工艺来说足够了,但源自这种传统CLD工作流程的细胞在灌注工艺中的性能可能无法维持,因为灌注工艺需要性能参数之间的复杂平衡。到目前为止,尚未对CLD工作流程对灌注工艺中顶级克隆体性能的影响进行全面研究。在这里,我们展示了CLD工作流程如何影响补料分批和灌注工艺中的细胞性能,强调了采用特定于灌注的CLD工作流程的优势,该工作流程包括使用专门为在灌注环境中扩增和生产而设计的培养基、更准确模拟灌注工艺的缩小模型,以及采用特定于灌注的细胞系选择标准。这些共同作用,导致开发出更高效的细胞系,适合连续和强化加工。