FEB 25, 2020 9:00 AM PST

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

  • Field Applications Scientist, Cell Therapy, Thermo Fisher Scientific
      Tia Hexom, PhD received a doctorate in cell and molecular biology at the University of Tulane in New Orleans, LA, USA. Dissertation work involved investigating cell morphology and synaptic plasticity of hypothalamic neurons during chronic variable stress and dehydration. Followed by a post-doctoral fellowship sponsored by the NIH Institutional Research and Academic Career Advancement (IRACDA) fellowship investigating chronic variable stress in Alzheimer's disease. Part of this work utilized iPSCs from Alzheimer's and control patients which were subsequently made into neural stem cells and utilized for the development of Alzheimer's disease in vitro modeling for future drug discovery screening. Dr. Hexom subsequently entered the cell therapy field and has gained extensive experience in the production of raw material blood products, GMP medical devices and translational workflows for CART and other cell therapy work. During this time Dr. Hexom has: led production labs isolating various immune cells and stem cells for cell therapy; managed medical device GMP manufacturing and developed marketing tools for application science and CART translational workflows. In her current role at Thermo Fisher Scientific, Dr. Hexom supports customers transitioning their cell and gene therapy work to clinical applications by providing workflow guidance and personalized scientific support.

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



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