Cryopreserved leukapheresis enables scalable and distributed CAR-T manufacturing: a multi-platform comparative study

Area of Science:

• Cellular immunotherapy
• Bioprocessing and manufacturing
• Hematology

Background:

• Cryopreserved leukapheresis is an underutilized but scalable source for Chimeric antigen receptor T (CAR-T) manufacturing.
• Current limitations include a lack of standardized preparation and low automation levels.

Purpose of the Study:

• To develop and validate a standardized, automated cryopreservation process for leukapheresis.
• To systematically evaluate the quality, functionality, and platform compatibility of cryopreserved leukapheresis for CAR-T production.

Main Methods:

• Optimized centrifugation, CS10 proportion, and cryopreservation procedures within a closed automated system.
• Evaluated post-thaw viability, cell recovery, phenotypic profiles, and functional CAR-T attributes.
• Assessed performance across non-viral, lentiviral, and Fast CAR-T platforms compared to fresh leukapheresis.

Main Results:

• Established a standardized cryopreserved leukapheresis process yielding ≥90% post-thaw viability.
• Cryopreserved leukapheresis showed comparable recovery and phenotype to peripheral blood mononuclear cells (PBMCs), with a higher lymphocyte proportion.
• Demonstrated comparable CAR-T cell viability, expansion, phenotype, CAR+ cell proportion, and cytotoxicity across different CAR-T platforms.

Conclusions:

• Cryopreserved leukapheresis is validated as a universal raw material for CAR-T manufacturing, preserving critical quality attributes and ensuring consistency.
• Standardization and automation enhance its utility, improving supply chain resilience by decoupling from fresh material logistics.
• Further large-scale clinical validation is recommended to solidify its role in CAR-T production.