Efficient automated processing for gene-engineered autologous cell therapies

摘要 基因工程自体细胞（GEAC）疗法治疗血液癌症和遗传性疾病是赚 越来越多的FDA批准，但生产效率低下（90%以上的废品率）导致长期 工作流程和高成本。目前的自动化工作并没有提供解决方案，因为细胞处理 他们自动化的技术-如基于Ficoll的细胞富集，磁性细胞分选（MACS），扩增和细胞 洗涤本身是低效的（就细胞回收而言）。本第1阶段SBIR将验证 显著更高效的电池处理过程（由申请人的功能封闭的X系列电池实现 处理盒和X-BACS基于浮力的细胞分离试剂，由康宁生命科学公司分销） 用于集成到“一锅式”GEAC制造工作流程中， 紧凑、价格适中的Quintessence仪器（后续第2阶段SBIR提案的主题）。 经证明，X系列检测盒可富集血液中的单核细胞，靶细胞回收效率为 90-100%（对于T细胞和造血干细胞），并以相等的回收率洗涤或体积减少细胞 效率同样，X系列检测盒中使用的X-BACS试剂的效率比传统检测盒高出50 MACS在靶细胞分离时的纯度非常高。结合起来，这些创新可以提供至少6倍的 相对于常规技术提高了细胞产率。由此制备的靶血细胞已经显示出 与常规产生的靶细胞一样容易地用慢病毒载体转导。 该提案的具体目标是：（1）原型和小批量生产的增强型X系列 具有支持后续目标所需的附加特征的药筒;以及（2）验证和优化 GEAC生产的所有步骤（从血液到转导、扩增和洗涤细胞）的性能 包括典型的治疗剂量）（“一锅”工作流程）， 第2阶段SBIR设计和验证Quintessence仪器，使工作流程完全自动化。 成功提供低成本、高效率、全自动化的GEAC制造技术， 通过使行业能够（1）制定这些疗法，大幅改善患者体验和结果 （2）降低GEAC的高制造故障率（无法生产完整的临床剂量）;（3） 减少从Rx到治疗的前置时间;（4）通过最小化离体细胞扩增产生更稳健的细胞;（5） 从白细胞去除术转换为外周血作为生产输入;以及（6）分散生产， 护理点（医院）。

ABSTRACT Gene-engineered autologous cell (GEAC) therapies for hematologic cancers and inherited disorders are earning growing numbers of FDA approvals, but manufacturing inefficiencies (90+% scrap rates) contribute to long workflows and high costs. Current automation efforts do not offer a solution, because the cell processing techniques they automate – like Ficoll-based cell enrichment, magnetic cell sorting (MACS), expansion and cell washing – are themselves inefficient (in terms of cell recoveries). This Phase 1 SBIR will validate the suitability of significantly more efficient cell handling processes (enabled by applicant’s functionally closed X-Series cell processing cartridge and X-BACS buoyancy-based cell isolation reagents, distributed by Corning Life Sciences) for integration into a ‘one-pot’ GEAC manufacturing workflow lending itself to full automation in the simple, compact, moderately priced Quintessence instrument (the subject of a follow-on Phase 2 SBIR proposal). X-Series cartridges are proven to enrich mononuclear cells from blood with target cell recovery efficiencies of 90-100% (for T cells and hematopoietic stem cells), and to wash or volume-reduce cells with equal recovery efficiency. Similarly, X-BACS reagents used in X-Series cartridges prove 50% more efficient than conventional MACS at target cell isolation at very high purity. Combined, these innovations can offer at least a 6-fold improvement in cell yield relative to conventional techniques. Target blood cells thus prepared have been shown to be transduced with lentiviral vectors as readily as are conventionally produced target cells. This proposal’s Specific Aims are: (1) to prototype and manufacture in small quantities an enhanced X-Series cartridge with additional features required to support the subsequent Aim; and (2) to validate and optimize the performance of all the steps of GEAC manufacturing (from blood to transduced, expanded and washed cells comprising a typical therapeutic dose) in a single such cartridge (‘one-pot’ workflow), the milestone for application for a Phase 2 SBIR to design and validate the Quintessence instrument that will fully automate that workflow. Success in delivering low-cost, high-efficiency, fully automated GEAC manufacturing technology may substantially improve patient experiences and outcomes by enabling the industry to (1) make these therapies more affordable; (2) reduce GEAC’s high manufacturing failure rate (failure to produce a full clinical dose); (3) reduce lead times from Rx to treatment; (4) produce more robust cells by minimizing ex vivo cell expansion; (5) switch from leukapheresis to peripheral blood as the manufacturing input; and (6) decentralize manufacturing to the point of care (hospitals).