Romero Juan J, Jenkins Eleanor W, Monroe Jacob I, Wickramasinghe S Ranil, Qian Xianghong, Bhattacharyya Dibakar, Husson Scott M
Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, South Carolina, USA.
School of Mathematical and Statistical Sciences, Clemson University, Clemson, South Carolina, USA.
Biotechnol Bioeng. 2025 Sep;122(9):2400-2409. doi: 10.1002/bit.29034. Epub 2025 May 29.
Technologies for large-scale manufacturing of viral vectors for gene therapies, such as tangential flow filtration and membrane chromatography, are under development. In these early stages of process development, techno-economic analyses are useful for identifying membrane properties yielding the greatest impact on process performance. In this study, we adapted a techno-economic framework used for monoclonal antibody capture for adeno-associated viral vector purification. We added mechanistic models to simulate flux decline during harvesting and separating full and empty capsids during polishing. Graphical user interfaces were added to help users explore the design search space. We selected a base process and manipulated selected variables to see their impact on large-scale manufacturing performance. These sensitivity analyses revealed that, under the selected process conditions, increasing module capacity reduces cost of goods more effectively than increasing operational flux in tangential flow membrane filtration modules for virus harvesting. Membrane chromatography columns with relatively low dynamic binding capacity (DBC) and short residence time (RT) offered similar or better economic performance than those with high DBC and long RT. Additionally, the difference in equilibrium solid-phase concentration between full and empty capsids as a function of salt concentration significantly affects purity.
用于基因治疗的病毒载体大规模制造的技术,如切向流过滤和膜色谱,正在研发中。在工艺开发的这些早期阶段,技术经济分析有助于确定对工艺性能影响最大的膜特性。在本研究中,我们将用于单克隆抗体捕获的技术经济框架应用于腺相关病毒载体的纯化。我们添加了机理模型来模拟收获过程中的通量下降以及在精制过程中分离完整和空衣壳的情况。添加了图形用户界面以帮助用户探索设计搜索空间。我们选择了一个基础工艺并操纵选定变量以观察它们对大规模制造性能的影响。这些敏感性分析表明,在选定的工艺条件下,对于病毒收获的切向流膜过滤模块,增加模块容量比增加操作通量更有效地降低了商品成本。具有相对较低动态结合容量(DBC)和较短停留时间(RT)的膜色谱柱比具有高DBC和长RT的膜色谱柱具有相似或更好的经济性能。此外,完整和空衣壳之间的平衡固相浓度差异作为盐浓度的函数会显著影响纯度。