Closed, Automated CAR-T Cell Manufacturing: from Research to Point of Care production.
- 1Limula
Abstract
Background: Point-of-care manufacturing of CAR-T cells could reduce cost and improve patient access, but few closed, automated systems support both flexible process development and GMP production. Methods and results: We describe LimCORE, a single-use consumable for cell incubation and processing, implemented in LimGROW (manual system) and LimONE (automated system). In LimGROW, T cells expanded 246 +/-18-fold over 10 days at optimized seeding density, versus 89 +/-9-fold in a G-Rex control, with similar viability (>90%) and phenotype. Process parameters directly transferred to LimONE, where automated, closed buoyancy-based CD3+ selection achieved 90.3 +/-3.5% purity and 59.2 +/-17.0% recovery in under two hours. Using this workflow, we manufactured CD19 CAR-T cells in an automated 7-day LimONE process requiring 155 minutes of operator intervention. LimONE manufactured CAR-T cells reached 42.6 +/-2.4-fold expansion (532 +/-29 M cells at harvest), with 43.6 +/-6.0% transduction efficiency and viability similar to G-Rex controls. CD4+/CD8+ ratios, CD8+ differentiation subsets, and cytotoxicity were comparable between platforms, while LimONE-manufactured cells showed increased metabolic fitness. Across CAR-T manufacturing runs on the LimONE, no hardware or software failures occurred, and automated liquid transfers and volume concentration stayed within +/-5% accuracy. Conclusions: These data show that LimCORE supports T cell expansion, selection, and complete CD19 CAR-T manufacturing with performance similar to existing platforms while reducing operator time, across R&D and GMP scale. Key words: Automation; CAR-T; Point of care; T cell manufacturing
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BibTeX
@misc{boudousquie2026closed,
title = {{Closed, Automated CAR-T Cell Manufacturing: from Research to Point of Care production.}},
author = {C. Boudousquie and D. Migliorini and J. Esensten and M. Irving and J. Maillard and R. Vuillefroy de Silly and Y. Pierson and V. Widmer and M. Pedard and M. Morbidelli and N. Vannini and E. Nallet and L. Bellosta and L. Dutoit and F. Jammes},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.08.748871},
url = {https://www.biorxiv.org/content/10.64898/2026.09.08.748871v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Closed, Automated CAR-T Cell Manufacturing: from Research to Point of Care production.
AU - C. Boudousquie
AU - D. Migliorini
AU - J. Esensten
AU - M. Irving
AU - J. Maillard
AU - R. Vuillefroy de Silly
AU - Y. Pierson
AU - V. Widmer
AU - M. Pedard
AU - M. Morbidelli
AU - N. Vannini
AU - E. Nallet
AU - L. Bellosta
AU - L. Dutoit
AU - F. Jammes
PY - 2026
DA - 2026/09/08
PB - bioRxiv
DO - 10.64898/2026.09.08.748871
UR - https://www.biorxiv.org/content/10.64898/2026.09.08.748871v1
AB - Background: Point-of-care manufacturing of CAR-T cells could reduce cost and improve patient access, but few closed, automated systems support both flexible process development and GMP production. Methods and results: We describe LimCORE, a single-use consumable for cell incubation and processing, implemented in LimGROW (manual system) and LimONE (automated system). In LimGROW, T cells expanded 246 +/-18-fold over 10 days at optimized seeding density, versus 89 +/-9-fold in a G-Rex control, with similar viability (>90%) and phenotype. Process parameters directly transferred to LimONE, where automated, closed buoyancy-based CD3+ selection achieved 90.3 +/-3.5% purity and 59.2 +/-17.0% recovery in under two hours. Using this workflow, we manufactured CD19 CAR-T cells in an automated 7-day LimONE process requiring 155 minutes of operator intervention. LimONE manufactured CAR-T cells reached 42.6 +/-2.4-fold expansion (532 +/-29 M cells at harvest), with 43.6 +/-6.0% transduction efficiency and viability similar to G-Rex controls. CD4+/CD8+ ratios, CD8+ differentiation subsets, and cytotoxicity were comparable between platforms, while LimONE-manufactured cells showed increased metabolic fitness. Across CAR-T manufacturing runs on the LimONE, no hardware or software failures occurred, and automated liquid transfers and volume concentration stayed within +/-5% accuracy. Conclusions: These data show that LimCORE supports T cell expansion, selection, and complete CD19 CAR-T manufacturing with performance similar to existing platforms while reducing operator time, across R&D and GMP scale. Key words: Automation; CAR-T; Point of care; T cell manufacturing
ER -