Akadeum’s Human T Cell Depletion Kit – CR is manufactured under strict adherence to GMP guidelines and tested to accepted standards for Ancillary Materials in Cell and Gene Therapy, with requirements for safety met.
In this workflow, unwanted T cells expressing CD3 are positively isolated using a cocktail of biotinylated anti-CD3 antibodies (CR), which are then captured using buoyant Human T Cell Depletion Microbubbles – CR 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.
Ancillary Materials Standards Adherence: ISO 20399, ISO 10993, USP <1043>
GMP Compliance per 21 CFR 820, 211, 210 and 11
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
Akadeum’s process offers an elegant solution for highly effective and scalable T cell depletion. Biotinylated anti-CD3 antibodies (CR) bind to untransduced T cells still expressing the CD3 epitope. Streptavidin coated Depletion Microbubbles – CR 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 – CR, leaving untouched engineered cells.
Additional Information:
Storage: 4°C
Kit components sterilized by filtration or electron beam irradiation
Go from research scale to commercialization with microbubbles. Just use more or less to suit your research needs and keep performance maintained.
Alerion™ Microbubble Cell Separation System compatibility coming in early 2025.
The Right Products to Support Your Pipeline:
With the appropriate safety testing completed, the Human T Cell Depletion Kit – CR is ideal for Phase I to commercialization of cell therapy workflows. GMP and RUO kits are also available for earlier stages of cell therapy pipelines.
Manufacturing by a third party, FDA-registered facility is available, please contact us.
Resources
User Guide
Learn how to integrate on your existing equipment:
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 and regulatory efforts.
Clinical Ready
Clinical Ready kits undergo additional testing beyond the requirements of GMP.
Akadeum is compliant with following standards for the Human T Cell Depletion Kit-CR:
ISO 20399 – Biotechnology – Ancillary materials present during the production of 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
The Human T Cell Depletion Kit – CR has 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)
The Human T Cell Depletion Kit – CR contains serum free and virus free antibodies, tested per ICH guidelines.
* “Use of International Standard ISO 10993-1, “Biological evaluation of medical devices – Part 1: Evaluation and testing within a risk management process” FDA Guidance, SEP2023
GMP
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.
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
Additional Regulatory Information:
Kit components are sterilized by filtration or electron beam irradiation. Akadeum is compliant with ISO 11137: Sterilization of Health Care Products – Radiation (2024)
Manufacturing by a third party, FDA-registered facility is available, please contact us.
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
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!
