Marshall Kurt, Mastro Melissa, Nankervis Brian, Shahid Shahid, Ciasullo Gabriella, Smith Trevor, Loveras Mary, Smith David, Miller Mindy M, Gibb Stuart L
Terumo Blood and Cell Technologies, Lakewood, Colorado, USA.
BioCentriq, Inc., Monmouth Junction, New Jersey, USA.
Cytotherapy. 2025 Mar;27(3):378-390. doi: 10.1016/j.jcyt.2024.11.003. Epub 2024 Nov 8.
Both quality of product and rapidity of manufacture are critical parameters if ex vivo manufacturing of autologous chimeric antigen receptor T cell (CAR-T) therapies is to reach its full potential. The Quantum Flex Cell Expansion System from Terumo Blood and Cell Technologies (Terumo BCT), a hollow-fiber bioreactor platform, is one of several cell expansion systems available to cell and gene therapy manufacturers to generate such cells in a GMP-compliant manner. In this study, the dynamic range of the Quantum Flex platform to expand CD19 CAR-T cells from variable quantities of starting material was investigated. Reflecting the industry's utilization of contract development manufacturing organizations (CDMOs) for accelerating clinical timelines, Terumo Blood and Cell Technologies performed a technology transfer of application protocols for study execution. Four different amounts of starting material (1, 3, 6 and 15 million cells) were expanded on Quantum Flex, using a unique donor's cells for each run. In this study, CAR-T cells were created using commercially obtained T cells and an anti-CD19 CAR-T lentiviral construct. The resultant heterogenous cell populations were expanded for 7 days in the functionally closed bioreactor platform. Expansion kinetics for all 4 starting material amounts were remarkedly similar, resulting in a 150- to 200-fold increase in cell numbers. This allowed for a study maximum of 2.6 billion cells from loading 15 million cells. Viability remained high throughout the expansion process with >93% for all 4 donors at harvest. To complete the manufacturing cycle, the automated and functionally closed Finia Fill and Finish System (Terumo Blood and Cell Technologies, Lakewood, CO) was used to formulate the cells for cryopreservation. Postprocedure analysis for potency and cytotoxicity demonstrated the production of efficacious cells. With this range of starting numbers, the platform is relevant to adult, pediatric and compassionate CAR-T expansion dosing. Today, several platforms are available to achieve sufficient cell yields for therapeutic applications of CAR-T, and awareness of the capabilities, pros and cons of each platform is critical to drive progress.
如果自体嵌合抗原受体T细胞（CAR-T）疗法的体外制造要充分发挥其潜力，产品质量和制造速度都是关键参数。泰尔茂血液与细胞技术公司（Terumo BCT）的Quantum Flex细胞扩增系统是一种中空纤维生物反应器平台，是细胞和基因治疗制造商可用于以符合GMP规范的方式生成此类细胞的几种细胞扩增系统之一。在本研究中，研究了Quantum Flex平台从不同数量的起始材料扩增CD19 CAR-T细胞的动态范围。为了反映该行业利用合同开发制造组织（CDMO）来加速临床进程的情况，泰尔茂血液与细胞技术公司对用于研究执行的应用方案进行了技术转移。使用Quantum Flex对四种不同数量的起始材料（100万、三百万、六百万和1500万个细胞）进行扩增，每次运行使用来自独特供体的细胞。在本研究中，使用商业获得的T细胞和抗CD19 CAR-T慢病毒构建体来创建CAR-T细胞。所得的异质细胞群体在功能封闭的生物反应器平台中扩增7天。所有四种起始材料数量的扩增动力学明显相似，细胞数量增加了150至200倍。这使得从接种1500万个细胞开始，最多可获得26亿个细胞用于研究。在整个扩增过程中活力保持较高水平，收获时所有4个供体的活力均>93%。为了完成制造周期，使用了自动化且功能封闭的Finia灌装和终产品系统（泰尔茂血液与细胞技术公司，科罗拉多州莱克伍德）对细胞进行配制以用于冷冻保存。对效力和细胞毒性的术后分析证明产生了有效的细胞。鉴于起始细胞数量的这个范围，该平台适用于成人、儿科和同情性CAR-T扩增给药。如今，有几种平台可用于实现CAR-T治疗应用所需的足够细胞产量，了解每个平台的能力、优缺点对于推动进展至关重要。
