245 Demonstration of a modular and affordable closed-system manufacturing process using the Sepax C-Pro and SMART G-Rex®100M-CS platforms
Year: 2026
Type: conference-abstract
Abstract: Background Automated CAR-T manufacturing platforms reduce contamination risks but restrict operators to high equipment costs and rigid reagent ecosystems, increasing production costs and facility footprints. To improve patient access and decentralized point-of-care manufacturing, we developed a modular, functionally closed-system process decoupling unit operations. Pairing the Cytiva Sepax C-Pro for T cell isolation/harvesting with SMART G-Rex ®100M-CS bioreactors maintains a strictly closed and sterile fluid path and allows flexible reagent sourcing while delivering high-quality CAR-T products comparable to those generated in G-Rex100M open systems. This integration optimizes instrument occupancy, providing a scalable decentralized framework to lower per-dose manufacturing costs.Methods Healthy donor leukapheresis T cells were enriched and resuspended in complete T-VIVO medium using the Sepax C-Pro NeatCell program. On Day 0, cells, activation reagent, and lentiviral vector were added to a SMART G-Rex ®100M-CS vessel through a closed transfer set. Medium was added by gravity fill on Day 3 to reach 1L. On Day 7, CAR-T cells were harvested via the GatheRex™, then washed and formulated using the Sepax C-Pro CultureWash program. Product specifications (CAR expression, viability, expansion, size, VCN, and RCL VSV-G) were evaluated per QC SOPs. Cell growth within the SMART G-Rex®100M-CS was monitored non-invasively using the SensoRex™ to interrogate the SMART sensor’s resonant frequency and compute the SMART Growth Index (SGI).Results High T-cell purity (>80%) with low residual B cells (<0.5%), monocytes (<5%), and NK cells (<7%) were achieved via Sepax C-Pro negative selection. Product viabilities remained high (>90%) with an average cell size >11 μm and >15-fold expansion, matching open G-Rex100M performance and confirming a functionally closed aseptic process. VCN was <5 copies per CAR + cells and residual VSV-G DNA remained below the limit of quantification. Curve fitting of cell concentration against SGI revealed a strong positive linear correlation (R2 > 0.99), confirming non-invasive SGI tracking accurately reflects cell density, where a baseline SGI of 5 reliably predicted a harvest yield of ≥ 1 × 109 CAR-T cells.Conclusions Our modular, closed-system process yields high-quality CAR-T products with critical quality attributes comparable to standard open protocols. Restricting Sepax C-Pro utility to Days 0 and 7 minimizes instrument occupancy, allowing a single unit to support multiple parallel patient runs. This decentralized model combines novel SMART sensor technology, closed system processing, flexible reagent sourcing, and offers a highly viable pathway to expand global CAR-T cell therapy access, especially for low- and mid-income countries lacking access to > Grade-B GMP facilities and capital for high-cost manufacturing platforms.Acknowledgements We are grateful to ScaleReady and Skroot Laboratory for their generous support and collaboration throughout these studies.
Language: English
Topic: CAR-T cell therapy research
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $2,490
OpenAlex ID: W7225139814