(441c) Flow-Based Membrane Technology to Engineer T-Cells

The use of chimeric antigen receptor (CAR) T cell therapies has become an important tool for the treatment of hematological cancers. Moreover, the FDA has recently approved six different CAR T cell therapies to treat said cancers. However, the manufacturing of CAR T therapies are a time-consuming multi-step process that requires isolation from the patient, activation, transduction, expansion, and autologous infusion. Due to the complex manufacturing steps, a single CAR T cell injection can cost a patient between $350k up to 450k. Therefore, there is a need to develop efficient and scalable processes to engineer T-cells in a cost-effective manner. Hydrogel coated membranes (HCM) are tunable biomaterials that can be functionalized with a wide variety of antibodies to achieve T-cell activation. Additionally, the use of tangential flow filtration (TFF) can improve transduction efficiency with a CAR. In this work, a combination of HCM and TFF was studied to improve T-cell activation and transduction.

In the present study, HCMs were prepared using a functional poly(ethylene glycol) hydrogel, which was selected due to its bioinert properties and easy scalability. HCM surfaces were modified with activating ligands (i.e., anti-CD3 and anti-CD28) for T-cell activation. Activation, exhaustion, and phenotype of primary CD3+ T-cells were characterized by flow cytometry. The comparison with industry standard TransAct found memory phenotype and minimal exhaustion with HCMs. Based on the activation experiments dynamic transduction experiments were performed on a custom TFF device using Jurkat and Primary CD3+ human cells. Using a range of flow and viral conditions, significantly higher transduction was achieved compared to the static control, resulting in a lower required concentration of infectious units per cell. In conclusion, the combination of HCM activation and TFF device transduction presented in this work have the potential to improve current CAR T production, leading to enhanced phenotypes, improved throughput, and scalability, increasing accessibility to patients.

Acknowledgements: This project was developed with an award from the National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL) and financial assistance from the U.S. Department of Commerce, National Institute of Standards and Technology (70NANB17H002).