Point-of-care manufacturing: Revolutionizing Cancer Treatment

MARS® Bar supports the core rapid CAR-T manufacturing process; for programs that also require freezing, storage, thawing, and surrounding process steps, Applied Cells and Tofflon provide a broader end-to-end workflow.

CAR-T cell therapy continues to expand beyond hematologic malignancies into autoimmune disease, solid tumors, and other emerging areas of cell therapy research. As the field grows, manufacturing remains one of the practical barriers to broader clinical translation.

Traditional CAR-T workflows can involve leukapheresis, PBMC preparation, target-cell isolation, activation, transduction, extended expansion, washing, formulation, and quality testing. Depending on the process, manufacturing alone may take 7–14 days or longer. The most direct way to address this challenge is to start with the core manufacturing process itself. Applied Cells developed the GoFast™ CAR-T workflow around MARS® Bar to shorten and simplify the path from starting material to CAR-T harvest. For programs that also require cryopreservation, controlled-rate freezing, storage, thawing, or additional surrounding process steps, the workflow can then be extended through the broader Applied Cells–Tofflon collaboration.

GoFast™ CAR-T: start with the core rapid manufacturing workflow

GoFast™ CAR-T was developed around a simple principle: rapid manufacturing requires redesigning the workflow, not merely compressing a conventional process. Rather than relying on prolonged ex vivo expansion, the workflow reduces preparation and handling steps and connects the key manufacturing operations into a shorter sequence. The process can begin directly from whole blood or smaller-volume apheresis material, reducing reliance on separate PBMC preparation and repeated centrifugation or washing. The core workflow centers on three major operations: T-cell isolation, rapid activation and transduction, and closed harvest and formulation of the CAR-T product. With approximately 24 hours of activation followed by approximately 24 hours of transduction and no prolonged expansion phase, the manufacturing cycle is designed to be completed within about 2–3 days. This creates a process foundation that can support rapid, decentralized, bedside, or near-bedside manufacturing models.

The GoFast™ workflow uses MARS® Bar for T-cell isolation and final wash/concentration, with only one user-selected peripheral instrument required for activation and transduction.

A closed, streamlined manufacturing approach

A rapid CAR-T process also depends on how cleanly the individual manufacturing steps connect. Within the MARS® Bar workflow, column-free magnetic separation, activation and transduction, short-term culture, and final CAR-T harvest can be linked through a closed fluidic pathway. MARS® Bar combines column-free separation, in-situ washing, and closed single-use tubing. Reducing transfers and open handling can help limit operator-dependent variability and contamination risk while creating continuity from cell isolation into downstream CAR-T processing.

Why column-free separation matters

Magnetic cell separation is a critical upstream step in many cell therapy workflows. In conventional packed-column systems, magnetically labeled cells enter a physical column, are retained in a magnetic field, and are then recovered through washing and elution. Performance can therefore be influenced by factors such as column structure, sample viscosity, cell concentration, flow rate, and elution conditions.
MARS® Bar takes a different approach. Its column-free design captures magnetically labeled cells within a flowing system, avoiding direct interaction between cells and column-packing material. This approach is designed to provide greater flexibility across different cell quantities, sample volumes, and process configurations.

Flexibility for different starting materials and bead strategies

The platform is designed to work with magnetic bead systems spanning approximately 50 nm to 4.5 μm, giving process developers flexibility to evaluate different bead strategies on the same instrument.
Not every CAR-T program begins with a standard leukapheresis volume. Early-stage research, translational studies, pediatric applications, and other specialized workflows may involve lower starting cell numbers or smaller sample volumes. MARS® can be adapted for inputs including small-volume apheresis samples, cord blood, low-cell-count materials, peripheral blood volumes of 50 mL or less, and other precious samples where minimizing transfer loss is important. Actual minimum sample volume and cell number depend on tubing configuration, cell type, and project-specific validation.

T-cell isolation performance in the reported workflow

Rapid manufacturing still depends on an efficient starting isolation step. In the data presented in the original Tofflon workflow article, MARS separation achieved 76% recovery. T-cell purity increased from 28% before separation to 96% after separation, while cell viability was reported at 94% after MARS processing compared with 71% before separation.

Example T-cell separation data showing recovery, enrichment in T-cell purity, and cell viability following MARS® processing.

Cell-composition analysis in the same article showed a MARS-processed fraction containing approximately 99% T cells, with low residual B cells, NK cells, monocytes, and neutrophils.

Comparison of starting-material cell composition with MARS®-processed and standard-workflow fractions, illustrating enrichment of the T-cell population.

Importantly, the workflow is designed to enable T-cell isolation directly from whole blood or apheresis material, reducing dependence on repeated centrifugation and separate PBMC-preparation steps.

When cryopreservation is needed: extend GoFast™ into a full end-to-end workflow

GoFast™ does not require every program to begin with a full cryopreservation chain. For teams working from fresh starting material, the core rapid manufacturing workflow can center on MARS® Bar and GoFast™ CAR-T. For programs that also require sample preservation or a broader set of upstream and downstream processing operations, Applied Cells and Tofflon are developing an expanded workflow around that core process. This extended configuration can connect sample collection with cryopreservation, controlled-rate freezing, liquid-nitrogen storage, thawing, cell processing, culture, formulation, and final product handling. Within the broader workflow, MARS® Bar performs key cell-separation and CAR-T harvest operations, while complementary Tofflon systems support surrounding steps such as freezing, storage, thawing, culture, and formulation. This allows sites to add cryopreservation and additional process infrastructure when their workflow requires it, rather than treating those steps as a prerequisite for rapid CAR-T manufacturing.

When cryopreservation is required, the GoFast™ core workflow can be extended with complementary Tofflon systems for controlled-rate freezing, cryogenic storage, thawing, culture, and formulation, while MARS® Bar remains the central platform for cell-selection and harvest operations.

A modular path to rapid CAR-T manufacturing

The central objective of GoFast™ is to simplify and shorten the CAR-T manufacturing process itself. MARS® Bar provides the core platform for rapid cell isolation and harvest, while the workflow can be expanded with additional technologies when a program requires cryopreservation, storage, thawing, or other surrounding manufacturing operations. This modular approach gives cell therapy teams a clearer starting point: implement the rapid GoFast™ process first, then build outward as site requirements evolve. For programs that need a more complete sample-to-product infrastructure, the Applied Cells–Tofflon collaboration provides a pathway to connect those additional steps into the same overall manufacturing strategy.

Interested in discussing rapid CAR-T manufacturing or how MARS® Bar and GoFast™ could fit into your workflow? Contact us to learn more.

Note: This blog post has been adapted from the original Tofflon WeChat article developed around the Applied Cells–Tofflon collaboration. The original technical flow, reported data, and figure sequence have been retained, while the text has been edited and reformatted for an English-language Applied Cells blog audience.

*For Research Use Only. Not for use in therapeutic or diagnostic procedures.

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