The Human T Cell Depletion Kit – GMP is manufactured under stringent adherence to GMP requirements. In this workflow, Human T Cell Depletion Microbubbles– GMP 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 (GMP), which are then captured using our buoyant Human T Cell Depletion Microbubbles– GMP 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.
GMP Compliance per 21 CFR 820, 211, 210 and 11.
Sterile, low endotoxin kit components per USP <71> and USP <85> guidelines.
Virus-free antibodies per ICH Q5A(R2).
Highly pure CD3 negative cells post-depletion.
Increased recovery of gene-edited cells for downstream formulation.
Gentle process maintains the viability of the cell therapy product.
Ancillary Materials for Cell & Gene Therapy: ISO 20399 and USP <1043> compliant.
Akadeum’s process offers an elegant solution for highly effective and scalable T cell depletion. Biotinylated anti-CD3 antibodies (GMP) bind to untransduced T cells still expressing the CD3 epitope. Streptavidin coated Human T Cell Depletion Microbubbles– GMP 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– GMP, 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
Kit components sterilized by filtration or electron beam irradiation
Storage: 2-8°C
Cell Type: T Cells
Sample Species and Source: Engineered Human CAR T cell cultures
Cell Separation Method: Positive (Depletion)
Capacity: From 10 x 106 to 5 x 109 T cells
This Kit Comes With:
5 Billion Cell GMP Kit:
50 mLs Human T Cell Depletion Microbubbles – GMP (in storage buffer)
5 mLs Human T Cell Depletion Antibody Cocktail – GMP
Go from research scale to commercialization with microbubbles. Just use more or less to suit your research needs and keep performance maintained.
The Right Products to Support Your Pipeline:
Manufactured in compliance with 21 CFR 820, the Human T Cell Depletion Kit – GMP is ideal for process development through commercialization. An RUO version is also available for early research.
Manufacturing by a third party, FDA-registered facility is available, please contact us.
Data
PBMC input material were prepared from washed apheresis cells and additional RBC lysis. Depletion was completed in accordance with the Human T Cell Depletion Kit – GMP protocol; isolations were performed in a BioRise™ Consumable using the Alerion™ Microbubble Cell Separation System. Averaged results from three unique donors comparing BioRise™ depletions to mid scale performed manually in a 150mL bag, respectively (depletion calculated based on remaining T cells in the negative fraction). Input cells for BioRise™ was 5 billion and input cells for 150mL bag was 1 billion. Average starting CD3+ cells was 49%.
Resources
User Guide
Learn how to integrate on your existing equipment:
For processing on the Alerion™, please refer to the:
App Note
Publications & References
Regulatory
As a supplier of ancillary materials for cell and gene therapy, Akadeum recognizes the criticality of confidence in materials and is committed to developing products with appropriate quality efforts.
Akadeum is compliant with following standards for the Human T Cell Depletion Kit-GMP
ISO 20399 – Biotechnology – Ancillary materials present during the production of the cellular therapeutic products and gene therapy products
ISO 10993 – Biological Evaluation of Medical Devices
USP <1043> – Ancillary Materials in Cell, Gene and Tissue Engineered Products
ICH Q3D(R2) Guideline for Elemental Impurities
BACS Streptavidin Microbubbles have been tested per the following biocompatibility tests (as determined in Table A.1: Biocompatibility Evaluation Endpoints, FDA, SEP2023*), reviewed by a certified toxicologist and have met specifications:
Cytotoxicity
Intracutaneous Reactivity
Acute Systemic Toxicity (IV and IP)
Hemolysis
Sensitization
Material Mediated Pyrogenicity
Thrombogenicity (with magnetics predicate)
Our GMP Grade products are tested per USP <85> Endotoxins and USP <71> Sterility prior to release by certified third-party labs. Functional testing includes cell isolation. Antibodies are virus free per ICH Q5A(R2) guidelines.
GMP standard adherence includes:
21 CFR 820
21 CFR 210
21 CFR 211
21 CFR 11
ISO 11137
Use of USP Class VI plastics in production and bottling
in addition to related ICH standards.
Kit components are sterilized by filtration or electron beam irradiation. Akadeum is compliant with ISO 11137: Sterilization of Health Care Products – Radiation (2024)
To request a GHS SDS or Certificate of Analysis, please contact us at info@akadeum.com
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
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!
