The Human T Cell Leukopak Isolation Kit – GMP was developed with BACS™ Microbubbles to isolate untouched T cells from washed leukopak material via negative selection, eliminating concerns of particles directly bound to T cells, and is manufactured under stringent adherence to GMP requirements. Leveraging the gentleness of flotation, non-T cells are targeted and removed with antibodies. Negatively selected T cells are suitable for a wide variety of downstream assays, including cell culture, flow cytometry, scRNA-Seq, engineering, or other functional studies. The Human T Cell Leukopak Isolation Kit – GMP has the capacity to isolate T cells from an average half-leukopak (12.5 x 10⁹ total cells).
The Human T Cell Leukopak Isolation Kit – GMP Grade is compliant with standards for Ancillary Materials in Cell & Gene Therapy: ISO 20399 and USP <1043>.
Key features:
GMP compliance per 21 CFR 820, 211 and 210.
Release testing: endotoxin per USP <85>, sterility per USP <71> and viral testing of antibodies per ICH Q5A(R2)
Biocompatibility and extractables testing completed per ISO 10993
Market-leading recovery, purity and viability results in more potent and persistent T cells
Less than 1 hour negative selection workflow contributes to cost savings up to 40% in a cell therapy workflow
How it Works: Cell health is preserved with closed system, in-bag, buoyancy based negative cell separation – truly untouched cells.
The Human T Cell Leukopak Biotin Antibody Cocktail – GMP is added to the leukapheresis product to label unwanted cell populations. BACS™ Streptavidin Microbubbles – GMP are added to the sample and bind to the biotin-labeled cells. Due to their inherent buoyancy, the microbubbles quickly and gently float the non-T cells to the surface. Weld the bag onto an existing system or vessel of choice to harvest and proceed with your preferred method of activation and expansion.
Additional Information:
Kit components sterilized by validated filtration or electron beam irradiation methods
Go from research scale to commercialization scale with microbubbles. Just use more or less to suit your research needs.
Microbubbles can scale from a billion cells in a tube to tens of billions in a cell culture bag.
Apheresis material was platelet washed using Thermo Fisher Scientific’s Rotea™ cell washing instrument and then isolated using Akadeum’s negative selection Human T Cell Leukopak Isolation Kit. Samples were processed at either 500 million total starting cells or 15 billion total starting cells. These assays were performed in triplicate.
Data
GMP Microbubbles Are the Path to Improved Therapies
Achieving high purity and yield is only part of the story. Fewer impurities lends to more effective engineering. With fewer impurities and less damaged cells post isolation, achieve up to 2x more engineered CAR-T cells at the final harvest, enabling a reduction in time and costs while maintaining healthier cells. At the end of the workflow, microbubble-based isolation leads to more potent T Cells, enabling more killing potential of the engineered T cells.
High Purity and Yield at Any Scale
Untouched T cells were isolated from platelet-washed leukopak material using the Human T Cell Leukopak Isolation Kit – GMP in accordance with the published protocol. Isolation was completed with either 100 million cells in a 5 mL tube or 1 billion cells in a 50 mL tube. Samples were run in triplicate. To determine T cell purity and yield, isolated T cells and an aliquot of the initial sample were labeled with CD45-FITC and CD3-APC Fire/750. The fluorescently labeled cells were analyzed by flow cytometry. Dead cells and debris were excluded from the analysis.
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.
With this in mind, Akadeum has engaged Dark Horse Consulting for regulatory support.
Akadeum’s regulatory compliance testing goes beyond the requirements of GMP.
Akadeum is compliant with following standards
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 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)
* “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
ICH Q5A(R2)
Use of USP Class VI plastics in production and bottling
in addition to other related ICH standards.
Additional Regulatory Information
Kit components are sterilized by validated methods for filtration or electron beam irradiation.
Akadeum is compliant with ISO 11137: Sterilization of Health Care Products – Radiation (2024)
To inquire about a Drug Master File, request a GHS SDS, 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!
