Data-driven experimental approach to increase transfection efficiency and prolonging vector production in transient systems

慢病毒载体（LVV）的生产对于体外基因修饰细胞疗法的出现至关重要，例如嵌合抗原受体T细胞（CAR-T）。然而，人们普遍认识到，在能力和能力方面都存在全球限制。LVV的关键制造挑战之一是转染过程的限制和延长生产时间。该项目专注于开发一种数据驱动的实验方法，以实现最佳生物工艺，与目前的金标准工艺相比，可以提高慢病毒（LV）转染效率和/或延长LV生产时间。该项目将采用的方法是将高通量实验研究的力量与基于机器学习的方法相结合，以确定提高转染效率的最佳因素。在对关键变量进行系统排序，建立设计空间后，进行实验研究，探索因子的设计空间。在整个实验工作过程中，将定期进行扩大规模的工作，以确认小规模研究和结果的可扩展性。此外，将进行调查，以评估上游优化对下游加工的影响。该项目具有显著增强和改善LVV生产的潜力，并确保为下一代先进治疗提供大量功能载体。该项目与EPSRC的“未来制造”研究主题密切相关，该主题侧重于研究和开发英国的高价值制造能力和产能。

The production of lentiviral vector (LVV) has been critical for the emergence of ex vivo gene-modified cellular therapies, such as chimeric antigen receptor T cells (CAR-T). However, it is widely recognised that there are global limitations on both capacity and capability. One of the key manufacturing challenges for LVV is the limitations around the transfection process and enabling prolonged production. The project is focused on developing a data-driven experimental approach to achieve an optimal bioprocess which results in higher lentiviral (LV) transfection efficiencies and/or prolonged LV production compared to the current gold standard process.The approach this project will adopt is to combine the power of high-throughput experimental studies in combination with machine-learning based approaches to identify the optimal factors for improved transfection efficiency. Following the systematic ranking of key variables and the establishment of the design space, experimental studies will be undertake to explore the design space of factors. Throughout the course of the experimental work, periodic scale-up work will be undertaken to confirm scalability of the small-scale studies and findings. Moreover, investigations will be undertaken to asses the impact the upstream optimisation on downstream processing.This project has potential to significantly enhance and improve the production of LVV and ensure that large quantities of functional vector are made available for the next generation of advanced therapeutics. This project is aligned closely to EPSRC's Manufacturing the Future research theme which focuses on researching and developing high-value manufacturing capability and capacity for the UK.