Akadeum’s Human T Cell Depletion Kit uses microbubbles to remove unedited CD3+ T cells from CAR T cultures. Unwanted T cells expressing CD3 are positively isolated using a cocktail of biotinylated anti-CD3 antibodies, which are then captured using our buoyant Human T Cell Depletion Microbubbles that gently float the targeted cells to the top of the sample. This leaves a highly pure sample of engineered cells untouched and ready for downstream formulation. The highly scalable nature of this process means that it can easily be incorporated into a wide variety of cell therapy manufacturing workflows.
Scalable to fit your cell therapy workflow needs.
Highly pure sample of CD3 negative cells post-depletion.
Increased recovery of gene-edited cells for downstream formulation.
Flexibility to integrate into many existing workflows.
Gentle process maintains the viability of the cell therapy product.
Available in two sizes: 1 Billion Cells and 5 Billion Cells.
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 coated Human T Cell Depletion Microbubbles then selectively bind to the antibodies labeling the undesired cells. Due to inherent buoyancy, the microbubbles naturally float to the surface of the culture vessel bringing along CD3+ cells. The bubble-bound cells can then be easily extracted off the surface with the depletion microbubbles, leaving untouched engineered cells.
Additional Information:
5 Billion cell kit is compatible with manual processing, in-bag workflows, and the Alerion™ Microbubble Cell Separation System
Storage: 4°C
Cell Type: T Cells
Sample Species and Source: Engineered Human CAR T cell cultures
Cell Separation Method: Positive (Depletion)
This Kit Comes With:
1 Billion Kit:
10 mL Human T Cell Depletion Microbubbles (in storage buffer)
1 mL Human T Cell Depletion Antibody Cocktail
5 Billion Kit:
50 mL Human T Cell Depletion Microbubbles (in storage buffer)
Go from research scale to commercialization with microbubbles. Just use more or less to suit your research needs and keep performance maintained.
*Only the 5 Billion Cell kit is compatible with the BioRise™
Experimental Overview:
PBMC input material was prepared from washed apheresis cells and additional RBC lysis. Depletion was completed in accordance with the Human T Cell Depletion Kit protocol; isolations were performed in a BioRise™Consumable using the Alerion™ Microbubble Cell Separation System. Averaged results from three unique donors comparing BioRise™ depletions (5B total cells) to small (50M total cells) and mid scale (1B total cells) performed manually in a 5mL tube or 150mL bag, respectively. Depletion was calculated based on remaining T cells in the negative fraction.
The Right Products to Support Your Pipeline:
The Human T Cell Depletion Kit is ideal for research through process development, with GMP and Clinical Ready versions available to support preclinical to commercialization of cell therapy workflows.
Data
Experimental Overview:
PBMC input material (5B total cells per donor) were prepared from washed apheresis material and additional RBC lysis. Depletion was completed in accordance with the Human T Cell Depletion Kit protocol; isolations were performed in a BioRise™ Consumable using the Alerion™ Microbubble Cell Separation System. The donor-specific CD3 depletion is shown with the average CD3 depletion (indicated by the dotted line). Depletion was calculated based on remaining T cells in the negative fraction.
Resources
User Guide
Learn how to integrate on your existing equipment:
For processing on the Alerion™, please refer to the:
App Note
Publications & References
FAQs
Cell Therapy Products
What are the minimum and maximum cell numbers that can be processed with Akadeum’s kits?
It is not recommended to process under 10 million cells per sample. The maximum number of cells processed is dependent on the size of the vessel as each kit is very scalable.
What selection method is used for Akadeum’s kits?
Akadeum’s Human T Cell Leukopak Isolation Kit is the first ever negative selection kit that can meet the scale requirements for cell therapy. The Human T Cell Depletion Kit also uses negative selection to deplete endogenous CD3+ T cells. The Human T Cell Selection, Activation, and Expansion Kit uses positive selection.
What is meant by washed leukopak material?
Washed leukopak material refers to a leukopak that has been processed to reduce platelets, exchange buffer, and concentrate the cells. This can be done manually via centrifugation or by using commercially available cell washing instruments.
Can unwashed leukopaks be used with Akadeum’s kits?
Yes, Akadeum has successfully processed raw leukopaks. Please email techsupport@akadeum.com to learn more.
Can Akadeum’s kits be used with cryopreserved samples?
Yes, Akadeum’s kits are designed to work with both fresh and cryopreserved samples.
Are Akadeum’s kits for autologous or allogeneic workflows?
Akadeum’s kits can be integrated into both autologous and allogeneic workflows.
General Questions
Are Akadeum microbubbles hollow silica spheres?
Yes, Akadeum’s microbubbles are hollow silica spheres. Silica is a material that has extensive precedence for use in therapeutics and diagnostics.
Are antibodies directly attached to the microbubbles?
Only specific kits, such as our Dead Cell Removal (DCR) and RBC microbubble kits, use antibody-coated microbubbles. Most Akadeum microbubble products are not coated with antibodies; instead, a biotinylated antibody cocktail is provided to use with the streptavidin-coated microbubbles.
What is buoyancy-activated cell sorting (BACS)?
BACS is Akadeum’s proprietary cell separation technology that uses buoyant microbubbles to isolate specific cell populations utilizing only the power of gravity.
What are microbubbles?
Akadeum’s microbubbles are hollow silica spheres that are on average 12 microns.
How big are microbubbles?
Microbubbles average 12 um in diameter. Learn more about microbubbles technology.
How many cells can a microbubble lift?
Microbubbles can carry as many as four cells.
What is the Separation Buffer?
Ca2+, Mg2+ free Dulbecco’s PBS containing 2mM EDTA, and 0.5% bovine serum albumin (BSA). BSA must be biotin free to not interfere with kit performance.
After sitting in the refrigerator, all the microbubbles have separated out of solution. Is there something wrong?
No, this is normal due to the buoyant properties of microbubbles. The solution will need to be mixed well before every use. If processing multiple tubes, mixing between tubes is necessary.
I don’t have an aspirator/vacuum available in my lab, can I still use Akadeum microbubbles?
Aspiration is the best and easiest way to remove the microbubble layer after centrifugation. Pipetting off the bubble layer is not very effective. Vacuum pumps are available from a variety of vendors.
How are Akadeum’s kits shipped?
Kits are shipped on ice packs with overnight delivery.
I have additional technical questions, who should I contact?
Please email all technical questions to techsupport@akadeum.com or check out our chatbot!
