FEB 25, 2020 9:00 AM PST

CAR-T Generation for Identity, Purity and Potency Assay Testing


DATE:  February 25, 2020
TIME:  9:00am PST, 12:00pm EST
Chimeric antigen receptor T-Cell (CAR-T) therapy has been approved by FDA to treat B-cell leukemias and has seen great clinical success. CAR-T cell manufacturing involves isolating T-cells from a patient, cell activation, engineering in the CAR construct, and expanding the cells to a therapeutic dose. To ensure patient safety and product quality, a number of quality control (QC) tests must be performed throughout the manufacturing process and for product release. These include confirming the identity, purity, and potency of the final CAR-T product. Often times, these assays are procured by multiple sources and may vary in performance resulting in inconsistent outcomes and product quality. To address these issues, CAR-T cells were generated in-house and used for the development and optimization of CAR-T specific characterization tools. T-cells were isolated from healthy donors, expanded in vitro, and transduced with a second generation (CD3ζ and 4-1BB) anti-CD19 CAR lentivirus. Using a clinically relevant workflow, expression of the CAR construct was obtained in a high, consistent manner. These CAR-T cells were then extensively characterized with pertinent analytical assays. In particular, PureQuant Methylation assay was used to measure percent of CD4/CD8 cells and also detect contaminating B cells and monocytes to high level of sensitivity. Our presented work flow details optimized methods of CAR-T Cell characterization that may be used in pre-clinical and clinical CAR-T manufacturing, following proper validation and regulatory approval.
Learning Objectives:
  • Show a small-scale CAR-T workflow using ThermoFisher products
  • Highlight characterization tools that can be utilized and applied in a CAR-T workflow
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FEB 25, 2020 9:00 AM PST

CAR-T Generation for Identity, Purity and Potency Assay Testing



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