CSL Behring Innovation GmbH, Marburg, Germany.
Biotechnol J. 2024 Mar;19(3):e2300348. doi: 10.1002/biot.202300348.
The development and manufacture of biopharmaceuticals are subject to strict regulations that specify the required minimum quality of the products. A key measure to meet these quality requirements is the integration of a sterile filtration step into the commercial manufacturing process. Whereas common procedures for most biologics exist, this is challenging for lentiviral vector (LVV) production for ex vivo gene therapy. LVVs nominal size is more than half the pore size (0.2 µm) of filters used for sterile filtration. Hence, highly concentrated virus solutions are prone to filter clogging if aggregation of viruses occurs or impurities attach to the viruses. Several filters were screened aiming to identify those which allow filtering highly concentrated stocks of LVVs of up to 1E + 9 transducing units mL , which corresponds to 4.5E + 12 particles mL . In addition, the effect of endonuclease treatment upstream of the purification process on filter performance was studied. In summary, three suitable filters were identified in a small-scale study (<15 mL) with virus yields >80% and the process was successfully scaled-up to a final scale of 100 mL LVV stock solution.
生物制药的开发和制造受到严格规定的约束，这些规定明确规定了产品所需的最低质量。满足这些质量要求的关键措施是将无菌过滤步骤集成到商业制造过程中。虽然大多数生物制品都有常见的程序，但对于体外基因治疗用慢病毒载体 (LVV) 的生产来说，这是具有挑战性的。LVV 的名义尺寸超过用于无菌过滤的过滤器孔径（0.2 µm）的一半。因此，如果病毒聚集或杂质附着在病毒上，高浓度的病毒溶液容易堵塞过滤器。筛选了几种过滤器，旨在鉴定那些能够过滤高达 1E + 9 转导单位 mL 的高浓度 LVV 库存的过滤器，这相当于 4.5E + 12 个颗粒 mL 。此外，还研究了纯化过程上游内切酶处理对过滤器性能的影响。总之，在小规模研究（<15 mL）中鉴定了三种合适的过滤器，病毒产率>80%，并且该工艺成功扩大到最终规模为 100 mL LVV 储备溶液。
