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.