连接临床前开发与临床生产：学术型 CAR-T 细胞治疗中慢病毒载体生产的转化 GMP 平台

The manufacturing of lentiviral vectors (LV) under Good Manufacturing Practices (GMP) remains a critical bottleneck limiting the clinical translation of academic CAR T-cell therapies. To address this challenge, we established and validated an integrated GMP facility (ViPro-IBiS-UPRC) for aseptic LV production within a public healthcare setting.

A stepwise optimization strategy was implemented to bridge preclinical development and GMP manufacturing, including refinement of transfection conditions, vector harvest timing, and scale-up surface transition. GMP-compliant production processes and quality control (QC) frameworks were subsequently developed and validated for HEK293T Lenti-X master and working cell banks (MCB/WCB) and for LV manufacturing.

Cell banks demonstrated high viability ( 94%), robust expansion capacity, confirmed identity by DNA fingerprinting, and absence of microbial and viral contaminants, including adventitious and endogenous retroviruses. GMP-produced LV batches achieved functional titers ranging from 9.95 10 7 to 3.07 10 8 TU/mL and fulfilled all release criteria, including sterility, absence of mycoplasma, endotoxins, and replication-competent lentivirus. Residual host cell DNA and protein levels remained within international regulatory predefined acceptance thresholds. Stability studies demonstrated preservation of vector functionality up to 12 months under cryopreservation conditions. Following regulatory inspection, ViPro-IBiS-UPRC obtained GMP certification from the Spanish Agency of Medicines and Medical Devices (AEMPS) for LV production, supporting the manufacture of CARTemis-1, an anti-BCMA CAR T-cell product under clinical evaluation in relapsed multiple myeloma patients.

Collectively, this work establishes a GMP-compliant academic platform for scalable LV manufacturing, enabling decentralized, cost-effective, and clinically compliant supply. This point-of-care manufacturing model strengthens the accessibility of academic CAR T-cell therapies within public healthcare systems. An academic GMP platform was validated for lentiviral vector production Preclinical LV optimization enabled efficient translation to GMP manufacturing Qualified HEK293T MCB and WCB met identity, safety and stability criteria Clinical-grade LVC1 batches fulfilled potency and release specifications The platform supports decentralized CAR-T manufacturing in public healthcare