The development of CAR-T cell preparations has revolutionized the treatment of plasma and B-cell malignancies. However, the high price limits the supply of approved CAR-T cell products to all patients. Another goal is clinical testing of new indications and innovative approaches such as the combination therapies. In-house production of CAR-T cells has been established for this purpose.

All process steps for generating CAR-T cell preparations from leukapheresis products are carried out on the CliniMACSProdigy platform in a functionally closed sterile disposable set. After 12 days, the final product is formulated after cell harvest. Transduction to CD19-specific CAR-T cells is achieved using a lentiviral expression vector (Lentigen Technology, Inc.). The quality of intermediate and end products is tested using validated methods for the following parameters: Identity, content, viability, purity, transduction efficiency (FACS), viral copy number (VCN, qPCR) and freedom from microbial growth (blood culture device), endotoxins (turbidimetric) and mycoplasma (qPCR). Compliance with predefined target ranges yields final products suitable for release.

The pharmaceutical quality system of the dept. of transfusion medicine was adapted to the regulatory requirements and extended to the new equipment and methods: qualifying instruments, extensive staff training and method validation. Except for the endotoxin assay, all tests were carried out by QC. Due to the identity with the IMP in an ongoing trial preclinical testing was not required. The production was successfully established in 6 production runs (1 without transduction) and in 4 aseptic process simulations. A median of 5.2*10e9 CD3+T cells and 2.2*10e9 CAR-T cells were obtained from 10e8 CD3+T cells seeded. The viability was 97.2%, the percentage of CD3+T cells 99.7%, the transduction efficiency 44.2%, the VCN 2.77. Absence of microbial growth, mycoplasma and endotoxins resulted in releasable products. Storage tests at 2-6°C proved product stability over 48 hours.

The consistent output of products ready for release demonstrates the successful implementation of an in-house CAR-T cell manufacturing. This enables clinical trials on innovative therapies and thus optimum patient care.

I hereby certify that, to the best of my knowledge, no aspect of my current personal or professional circumstance places me in the position of having a conflict of interest with this presentation.