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)
