Akadeum’s Human T Cell Depletion Kit was designed to address an unmet need for purifying CAR T cells for cell therapies. This solution surpasses purity requirements, removing over 95% of CD3-positive cells while returning nearly all of the engineered cells for downstream use. Akadeum’s T Cell Depletion kits are designed to scale with the needs of therapy developers. Whether processing 10 million or 5 billion cells, these kits are perfect for any workflow. In addition, the gentleness of flotation-based separation results in healthier cells for more effective therapies. In all, Akadeum’s novel approach to CAR-T cell purification offers multiple improvements over current purification methods and the flexibility of Akadeum Microbubbles allows for rapid optimization of cell therapy workflows.
Mechanism of Action: When developing CAR T cell therapies, the ability to isolate pure populations of engineered cells after transduction is paramount. Cells still expressing CD3 can be harmful to the patients receiving these therapies. The Human T Cell Depletion Antibody Cocktail recognizes the CD3 epitope using multiple, non-overlapping anti-CD3 clones for enhanced performance. CD3+ cells are labeled with antibodies, which are bound by the BACSTM Depletion Microbubbles and removed from the sample using buoyancy. Unlabeled and successfully transduced CD3-cells are recovered by removal of microbubble-bound CD3+ cells, ready to use for downstream applications.
How it Works: Akadeum’s process offers an elegant solution for highly effective and scalable T cell depletion. Biotinylated anti-CD3 antibodies bind to untransduced T cells still expressing the CD3 epitope. Streptavidin microbubbles then selectively bind to the antibodies labeling the undesired cells. Due to the natural buoyancy of the microbubbles, they naturally float to the surface of the culture vessel pulling along CD3+ cells. The positive cells can then be easily extracted off the surface with the microbubbles leaving engineered cells untouched to settle to the bottom of the vessel.
- Storage: 4°C
- Cell Type: T Cells
- Sample Species and Source: Engineered Human CAR T cell cultures
- Cell Separation Method: Depletion
- Capacity: From 10 x 106 to 5 x 109 T cells
This kit comes with:
- 10 mL BACSTM Depletion Microbubbles in storage buffer
- 1 mL Human T Cell Depletion Antibody Cocktail in PBS
Recommended for Best Results
- 20 rpm tube rotator for mixing (e.g., Thermo Scientific cat#: 88881002)
- Centrifuge (swinging bucket rotor strongly preferred)
- Vacuum aspirator
- 9-inch sterile Pasteur Pipets
- Sterile 5 mL / 50 mL tubes
- Buffer of choice: Phosphate-buffered saline (PBS, Ca2+ and Mg2+ free) supplemented with 0.5% serum and 2 mM EDTA, pH 7.2 is recommended.