High Efficiency Microfluidic Device for Large Scale CAR-T Cell Manufacturing

项目摘要/摘要 该项目的目标是开发一种用于浓缩白细胞的高产量微流控设备 (WBC)来自白细胞分离产品，用于制造CAR-T和其他癌细胞 免疫疗法。CD19靶向CAR-T细胞疗法非常有效，FDA批准用于治疗 晚期恶性血液病，目前有300多项活跃的临床试验正在使用CAR-T细胞 针对各种癌症，包括实体瘤。大规模生产细胞免疫疗法， 包括CAR-T细胞疗法，构成了一个独特的挑战，因为每种疗法都是定制的- 由癌症患者自己的白细胞制成，使用白细胞分离法收集。因为癌症 患者通常有低白细胞计数和不稳定的外周血细胞访问，他们的白细胞采集是 通常白细胞数量有限，红细胞(RBCs)污染严重。有一个 用于CAR-T细胞制造的WBC来源材料的高产量浓缩的迫切需要。CG Science正在开发一种具有成本效益的设备，可以高产率地浓缩WBC，基于其 专利微流控“高效确定分离(HEDS)”技术。技术 旨在(1)提供对细胞癌免疫治疗至关重要的高WBC回收率，以及(2) 克服了其他微流控细胞分离的成本和吞吐量问题 来自成功商业化的技术。初步数据表明，HEDS有能力 提供&gt；95%的WBC恢复和&gt；98%的RBC去除，同时细胞活力损失无法检测- 显著优于现有的WBC浓缩系统。HEDS的独特配置可以 潜在地提供可扩展的处理吞吐量、抗堵塞能力和操作能力 重力进料。为了进一步确定HEDS技术用于电池制造的可行性，本第一阶段 该项目将重点关注3个具体目标：(1)优化HEDS芯片设计，开发芯片组件以实现规模化 加工吞吐量，(2)表现出高的WBC回收率，并表征浓缩的WBC 用流式细胞仪和T细胞增殖试验检测T细胞亚群，以及(3)显示塑料疱疹 芯片可制造性采用软压花法。研究小组包括HEDS的发明者 技术、免疫学生物学家、塑料设备制造工程师和 专门从事细胞治疗和细胞制造。这个项目的成功将带来丰收。 WBC浓缩装置，将显著改善细胞免疫疗法的来源材料， 使电池制造过程更加稳健和可靠，并最终提供更强大和 性价比高的靶向癌症治疗将使许多患者受益。

Project Summary/Abstract The goal of this project is to develop a high-yield microfluidic device for enriching white blood cells (WBCs) from leukapheresis products, for use in manufacturing CAR-T and other cancer cellular immunotherapies. CD19-targeted CAR-T cell therapy is highly effective and FDA-approved for treating advanced hematologic malignancies, and over 300 active clinical trials are currently using CAR-T cells to target various cancers, including solid tumors. Large-scale production of cellular immunotherapy, including CAR-T cell therapy, poses a unique challenge due to the fact that each therapy is custom- manufactured from the cancer patient’s own WBCs, collected using leukapheresis. Because cancer patients often have low WBC counts and inconsistent peripheral access, their leukapheresis harvests are frequently limited in WBC quantities and highly contaminated with red blood cells (RBCs). There is a critical need for high-yield enrichment of the WBC source material for CAR-T cell manufacturing. CG Scientific is developing a cost-effective device that enriches WBCs with high yields, based on its patented microfluidic “High Efficiency Deterministic Separation (HEDS)” technology. The technology aims to (1) provide high WBC recovery that is essential to cellular cancer immunotherapy, and (2) overcome the cost and throughput issues that have precluded other microfluidic cell separation technologies from successful commercialization. Preliminary data have shown that HEDS is capable of delivering >95% WBC recovery and >98% RBC removal, with undetectable loss of cell viability— significantly outperforming existing WBC enrichment systems. The unique configuration of HEDS can potentially provide scalable processing throughput, resistance to clogging, and capability to operate by gravity feed. To further establish the feasibility of HEDS technology for cell manufacturing, this Phase I project will focus on 3 Specific Aims: (1) optimize HEDS chip design and develop Chip Packs to scale up processing throughput, (2) demonstrate high WBC recovery yields and characterize the enriched WBCs and T cell subsets using flow cytometry and T cell proliferation assay, and (3) demonstrate plastic HEDS chip manufacturability using soft embossing. The investigator team includes the inventor of the HEDS technology, immunology biologists, plastic device manufacturing engineers, and medical doctors who specialize in cellular therapy and cell manufacturing. The success of this project will lead to a high-yield WBC enrichment device that will significantly improve the source material for cellular immunotherapies, make the cell manufacturing process more robust and reliable, and eventually provide more potent and cost-effective targeted cancer therapies that will benefit many patients.