Establishing a GMP-compliant manufacturing process and phase-appropriate analytics for early development of a FiCAR T-cell product with a novel CAR spacer

There is a growing demand for chimeric antigen receptor (CAR) -T cells for clinical trials. Consequently, new centers capable of manufacturing advanced therapy medicinal products (ATMPs) are needed. In this study, we established a good manufacturing practice -compliant manufacturing process and phase-appropriate analytics for a novel autologous CD19-targeted CAR T-cell product, 19-FiCART. We evaluated the stability of fresh, healthy donor-derived leukapheresis products (LPs), produced 19-FiCART using a 12-day semi-automated process with CD4/CD8-positive cell enrichment and lentiviral transduction, and evaluated the in vivo efficacy of 19-FiCART in a xenograft mouse lymphoma model. The optimal hold time and temperature to maintain LP stability were up to 73 h at 2-8 °C. The 19-FiCART manufacturing process consistently yielded more than 2 × 10⁹ highly viable CAR+ T cells, which is considered sufficient for a clinical product. The 19-FiCART products also demonstrated potent anti-tumor activity both in vitro and in vivo. This paper provides a detailed description of the manufacturing process and analytics for 19-FiCART and provides insights into the development of a release strategy for novel CAR T-cell products intended for early clinical studies. Additionally, we present data on LP stability, which has broader implications for the development of various immune cell-based ATMPs.