Formulation Considerations for Autologous T Cell Drug Products

Abstract

Genetically modified autologous T cells have become an established immunotherapy in the fight against cancer. The manufacture of chimeric antigen receptor (CAR) and αβ-T cell receptor (TCR) transduced T cells poses unique challenges, including the formulation, cryopreservation and fill–finish steps, which are the focus of this review. With an increasing number of marketing approvals for CAR-T cell therapies, comparison of their formulation design and presentation for administration can be made. These differences will be discussed alongside the emergence of automated formulation and fill-finish processes, the formulation design space, Monte Carlo simulation applied to risk analysis, primary container selection, freezing profiles and thaw and the use of dimethyl sulfoxide and alternative solvents/excipients as cryopreservation agents. The review will conclude with a discussion of the pharmaceutical solutions required to meet the simplification of manufacture and flexibility in dosage form for clinical treatment.

Increasing Gene Editing Efficiency via CRISPR/Cas9- or Cas12a-Mediated Knock-In in Primary Human T Cells

“…An excessively high level of expression from a strong promoter can disrupt the functional properties of cells or expression may become silenced [45]. Third, due to the random integration site, the cell product is highly heterogeneous and varies from donor to donor [8,46,47]. This makes it difficult to analyze results of clinical trials and predict the effectiveness of therapy for a given patient.…”

“…One important consideration for the production of cells with genome editing is the percentage of cells with the desired modification that would be enough to achieve a therapeutical benefit. Interestingly, the level of transduction in CAR T or TCR T cell products varies from 2% to 96%, leading to different numbers of modified T cells being infused into a patient [8,46,47]. The level of transduction correlates with the differing number of vector copies per cell (vector copy number, VCN) [57].…”

Streamlined Methods for Processing and Cryopreservation of Cell Therapy Products Using Automated Systems

“…Clinical laboratories routinely cryopreserve cells using cryopreservation solutions and operating conditions that need to be tightly controlled and amenable to the cell type for optimal cell viability and yield. The use of automated equipment that combines multiple manufacturing processes, such as fill-finish systems and controlled-rate freezers, can offer several advantages over manual operations including automation, better control of processing, and efficiency, thereby avoiding potential compromises to the viability and yield of cell products ( van der Walle et al, 2021 ). Together, these systems can be employed and programmed to optimize the operations for cell cryopreservation at end-stage manufacturing as well as starting materials upstream of manufacturing.…”