CAR-T Lentiviral Vector Expression Testing, Identity and Purity Assessment Throughout the Cell Therapy Manufacturing Process
A COLLABORATION BETWEEN ACCELLIX, NECSTGEN AND BIOMÉRIEUX
INTRODUCTION
Cell and gene therapies (CGT) have the potential to provide a treatment and potential cure for thus far uncurable diseases. Manufacturing of these therapies like adoptive T cell therapies, such as TCR and CAR-T cells, include selection of starting population, cell activation, transduction, expansion and fill and finish of the product. Quality control is a vital aspect of manufacturing. Process controls closely monitor the production process, while release tests assess the required quality of the final product. Quality control tests for CAR-T cell therapy products include tests for identity and purity. An accepted technique for testing identity and purity is flow cytometry, which assesses the T cell content and target receptor expression in the final product. Quality control tests must comply with regulations and guidelines to make sure the tests are fit for purpose and suitable for the phase of CGT manufacturing. Additionally, time-constraints for testing should be considered, since patients may be in urgent need of the therapy.
In this work, we directly compared flow cytometry analysis of two flow cytometry systems using a CAR-T cell production workflow (Figure 1), where the T cells were transduced with an in-house manufactured lentiviral vector to express enhanced green fluorescent protein (eGFP) as a reporter target gene.
We show that the Accellix Platform, a cartridge-based benchtop flow cytometry platform with algorithm-based automated analysis, is a reproducible alternative to other flow cytometers for CGT quality control testing as well as early cell and gene therapy manufacturing.
In this technical note, we use the Accellix automated cartridge-based flow cytometry platform to measure the transduction efficiency of a lentivirus vector containing eGFP as a reporter gene. In collaboration with NecstGen, a non-profit CDMO and center of excellence for cell and gene therapy, and bioMérieux, we demonstrate the use of the Accellix Platform in the cell therapy manufacturing process.
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