T Cell Manufacturing For Cell Therapy
T cell-based therapies hold significant potential for treating intractable cancers by redirecting the power of living immune cells. These engineered therapies take advantage of the inherent functions of specific T cell types associated with fighting disease. Continuing technological advances in manufacturing as well as deeper biological understanding will improve the performance and accessibility of these cutting-edge cell therapies.
From ancillary materials to automated analytical tools, Bio-Techne is committed to delivering innovative solutions that enable cell and gene therapies to reach more patients.
Use the icons below to navigate to products and services for each step of the T cell manufacturing process.
As Your Program Advances Towards the Clinic, We Will Grow With You
Serum-Free and Animal-Free Cell Culture
Serum-Free and Animal-Free Cell Culture
Increase the consistency of your cell cultures as you approach translational studies for cell therapy programs. Adopting these media will:
- Reduce variability in media composition
- Simplify compliance with regulatory guidelines
- Streamline comparability testing for raw material changes
GMP Capabilities
GMP Capabilities
We strive to reduce your manufacturing risks by providing industry-leading quality, consistency, reliability, and capacity.
- Quality compliance and certifications
- Custom services
- Animal-free manufacturing
- Our GMP facilities around the world
Transitioning From Research to GMP
Transitioning From Research to GMP
As you advance from discovery into process development and clinical manufacturing, choose GMP raw materials with the equivalent performance as the research-grade products you use. This will make your transition as efficient and seamless as possible.
T Cell Isolation
Begin your immunotherapy process on a strong footing. Accurate, efficient, and reproducible isolation of T cell populations will lay the groundwork for a successful cell therapy program. Rigorous analysis of your cell populations for phenotype and viability as well as the lack of contaminating cell types is critical to confirm that your isolation procedure is effective.
T Cell Selection
T Cell Selection
Accurate and reproducible cell selection depend on the specificity of the antibodies you choose. Your isolation of high purity T cell populations will be more reliable when you choose the right antibodies for the purpose.
- Recombinant antibody engineering and GMP antibodies
- Monoclonal panel development
- Specialty conjugation
T Cells From Induced Pluripotent Stem Cells (iPSCs)
T Cells From Induced Pluripotent Stem Cells (iPSCs)
T cells differentiated from iPSCs support the development off-the-shelf, allogeneic cell therapies. The reduced costs and more rapid therapy development with universal cell banks can make cell therapies available for treating more patients.
T Cell Culture and Activation
Expand your T cell cultures to a clinically effective scale with optimized growth media and equipment. Your ancillary materials including media and supplements should be standardized for activity and reproducibility. This will boost cell culture consistency and confirm the accuracy of your cytokine additions with specific immunoassays.
GMP Human T Cell Media
GMP Human T Cell Media
Improve the consistency of your T cell cultures with a clinic-ready media. This media enables robust T cell expansion from PBMCs or purified T cells.
Quantitate Secreted Cytokines
GMP Cytokines & Growth Factors
GMP Cytokines & Growth Factors
Supplement your T cell expansion cultures with Animal-Free and GMP Proteins including IL-2, IL-7, and IL-15. Choose GMP cytokines with bioactivity that is benchmarked against research-grade cytokines to minimize process disruption during the transition. Confirm accurate cytokine addition with precise and reliable ELISA kits.
T Cell Activation
Assess T cell activation markers rapidly and precisely with Simple Plex assays on the Ella platform to boost cell therapy development and manufacturing efficiency.
Granzyme B | IFN-γ | IL-2 | Perforin | TNF-α
T Cell Exhaustion
For effective T cell therapies, it is important to avoid overactivation of the cells which can lead to T cell exhaustion. This balance may be dependent on the affinity and number of interactions between the T cell therapy and its target cells. Verify the functional potential of your T cells by analyzing the expression of exhaustion markers such as PD-1 and CTLA-4.
Bio-Techne believes in the power of partnerships. We have joined with Fresenius Kabi and Wilson Wolf to bring you ScaleReady™, lowering the barriers to entry into the immune cell therapy space for all sizes and stages of manufacturing programs. ScaleReady brings cell and gene therapies to life by offering a significant reduction of complexity and cost while consistently providing superior repeatability and cell quality.
The ScaleReady product portfolio supports scalable low/no-touch T cell activation and expansion, delivering true platform, process, and product continuity for your T cell programs. When paired with the Lovo® and Cue® functionally-closed-system automated cell processing systems, Bio-Techne reagents and the G-Rex® Bioreactor enable high throughput parallel processing of cell therapies within a small footprint.
TcBuster™ Transposon System for Non-Viral Gene Delivery
Engineer your T cells more quickly and easily by using non-viral gene delivery. TcBuster will fast-track your path to the optimum cell therapy phenotype.
- Reduced cost of genetic modification
- Rapid cell engineering with manufacturing controls
- Integration of larger DNA cargoes
- Flexibility to deliver multiple genes in one operation
- Unaffected by lentivirus vector supply shortages
Flow cytometry analysis of CD19 CAR expression in human T cells following introduction with TcBuster. Cells were analyzed by staining with Recombinant Human CD19 Protein, Atto 647N Conjugate. Histograms represent analysis of T cells from a single donor in triplicate (blue, red, green) and a no-transposase control (black). TcBuster showed an average transposition efficiency of 65.4%.
T Cell Engineering With Viral Vectors
Vector characterization
- Capsid characterization with iCE Maurice™
- Capsid monitoring during purification with Simple Western™
- Identification of capsid aggregates and contaminants with Micro-Flow Imaging™
Lentiviral vector titration with HIV-1 Gag p24 ELISAs and Reagents
Automated AAV viral titer assays developed in partnership with PROGEN
Process efficiency improvements with viral transduction enhancers
Confirming Successful Cell Engineering
Confirm that your T cells have the phenotype that reflects the gene editing you performed. Characterize their surfaces for expression of CARs and accessory proteins as well as their secretory profiles for cytokine production.
Human CD4+ and CD8+ T cells were either (A) transduced or (B) not transduced (negative control) with a human CD19 CAR and then cultured for 11 days. Cells were stained with Human CD4 PE-Cy7-conjugated Antibody and Recombinant Human CD19 Atto 488-conjugated Protein.
Evaluating CAR Expression on CAR-T Cells Using FluorokinesTM
Evaluating CAR Expression on CAR-T Cells Using FluorokinesTM
One of the current challenges in the field of CAR-T cell therapy is the lack of a simple assay to easily detect and quantify CAR expression following T cell transduction. Read this application note to learn about the advantages of using R&D SystemsTM Fluorokines fluorescent-labeled proteins to directly stain and detect CAR+ cells by flow cytometry.
The CAR ToolKit: Progress and Challenges in CAR-T/NK Cell R&D and Mfg
The CAR ToolKit: Progress and Challenges in CAR-T/NK Cell R&D and Mfg
With numerous clinical trials underway and multiple CAR-T cell products now approved, solving the current obstacles when manufacturing CAR-T/NK cell products continues to be a pressing issue for the cellular immunotherapy field. Watch this webinar, hosted by BioInsights, to learn about the current challenges, latest progress, and innovation facing CAR-T and CAR-NK cell development.
T Cell Characterization
Final T cell product characterization should encompass all your defined critical quality attributes (CQAs) to ensure that your product is as safe as possible for in vivo administration. Orthogonal CQA testing methods analyze unrelated parameters of the product and include phenotypic characterization of the cells, secretory profile, and purity (e.g. lack of undesired cell types and particulates).
T Cell Phenotype
Characterize the cell surface receptors on your T cells thoroughly to determine if they exhibit the correct phenotype for the cell type and activation status you need. In addition, measure population heterogeneity to determine if the count of contaminating cells is low enough to meet your specifications.
Human Th1 cells were stained for CD4 expression using CD4-Alexa Fluor 700 followed by IFN-gamma-PerCP, IL-12 Rβ2-PE, and T-bet-Alexa Fluor 488 antibodies. Cells were fixed and permeabilized with the FlowX™ FoxP3 Fixation & Permeabilization Buffer Kit. Flow cytometry quadrants were set based on staining with isotype controls (Catalog # IC003N, # IC006T, # IC002P, # IC0041C, and # IC002G).
T Cell Secretory Profile
(A) Quantitation of human IFN-gamma with the Simple Plex and Quantikine™ HS ELISA. Performance of the two immunoassay platforms are highly correlated (R2=0.99).
(B) Multianalyte cytokine and chemokine quantitation with the Human Immunotherapy Magnetic Luminex Performance Assay 24-plex Fixed Panel. Linearity of detection and spiked recovery of each analyte is 3-4 logs in a serum matrix. B7-H1, CCL2, CCL3, CCL4, CD40 Ligand, CXCL10, GM-CSF, Granzyme B, IFN-alpha, IFN-gamma, IL-10, IL12p70, IL-13, IL-15, IL-17A, IL-1ra (BYU), IL-1 beta, IL-1ra, IL-2, IL-33, IL-4, IL-6, TNF-alpha.
Contaminant Screening
Analyze the morphology and quantitate the levels of particulates, residual cell activation beads, and other impurities in your T cell product. Make sure they are low enough to meet your requirements for in vivo administration.
- Micro-Flow Imaging™
- Simple Western™ - Screen for host cell proteins and bioprocess contaminants with high sensitivity immunoassays and total protein detection.
- Simple PlexTM automated HEK HCP 3G process residual assay developed in partnership with Cygnus Technologies
Distinguish between particle types in multiple tandem samples with MFI. (A) MFI Image Analysis software filters were set for Jurkat T cells and Dynabeads, where ECD is the equivalent circular diameter and Intensity Std is the standard deviation of the intensity of all pixels of that particle. (B). MFI distinguishes between the two particle types within a mixed population.
Tissue Biopsy and T Cell Localization
Monitor your therapy after administration by analyzing T cell integrity and trafficking with single-cell resolution. In situ analysis in the context of intact tissue morphology provides detailed information about infiltration and persistence of the T cell therapy in the tumor.
- RNAscope™ and BaseScope™ assays
- Good Clinical Laboratory Practice (GCLP)-compliant RNAscope™ ISH analysis
- Professional Assay Services
The RNAscope Multiplex Fluorescent V2 assay was combined with immunofluorescence (IF) to visualize tumor infiltration of activated anti-BCMA CAR-T cells. RNAscope ISH detection of the 3' UTR of the CAR vector (green), Granzyme B (red), and IFN-gamma (pink) was followed by IF for CD3 (white) in xenograft tumors in RPMI-8226 mice.
In Vivo Host Monitoring
Measure the host response to your T cell therapy with R&D Systems Luminex® Assays. The Human XL Cytokine Performance panel profiles up to 46 targets simultaneously. Find the format that meets your needs.
Custom Services
Your cutting edge cell therapy should never be delayed from reaching patients just because the right materials aren’t available. If you need specially designed reagents or assays for developing your T cell therapy, we will work with you to deliver optimized solutions for streamlining your process. We will gladly provide customized formulation and packaging as well as bulk orders.
Optimizing raw materials to fit your process requirements can provide significant benefits in efficiency, safety, and cost. Customization is important to do early in the process, and it takes on increasing importance as a therapy gets closer to regulatory approval. It is critical to identify a supplier that can function as a flexible partner to enable you to standardize your process at scale and with consistency. With Bio-Techne you will find a partner that is readily accessible to provide technical and regulatory support throughout your process.
This new eBook provides an overview of several biological and manufacturing challenges facing the development and production of T cell therapies. It follows with examples of how Bio-Techne products and services can address these challenges at each stage of the process.
As you develop and evaluate new targets for T cell therapies, these tools will help you to characterize important molecules in even greater detail.
| targets for NK cell therapy | Antibodies | ELISA Kits | Simple Plex | Proteins | Luminex | Proteome Profiler |
| alpha-Smooth Muscle Actin | Yes | |||||
| B7-1/CD80 | Yes | Yes | Yes | |||
| B7-2/CD86 | Yes | Yes | ||||
| B7-H3 | Yes | Yes | Yes | Yes | ||
| BCMA | Yes | Yes | Yes | Yes | Yes | |
| Cadherin 17 | Yes | Yes | ||||
| CD7 | Yes | Yes | ||||
| CD19 | Yes | Yes | ||||
| CD20 | Yes | |||||
| CD27 Ligand | Yes | Yes | Yes | Yes | ||
| CD30 | Yes | Yes | Yes | Yes | Yes | |
| CD34 | Yes | Yes | Yes | |||
| CD38 | Yes | Yes | Yes | |||
| CD117/c-Kit | Yes | Yes | Yes | Yes | Yes | |
| CD160 | Yes | Yes | Yes | |||
| EGFR | Yes | Yes | Yes | Yes | Yes | |
| EGFR Viii | Yes | |||||
| EMMPRIN/CD147 | Yes | Yes | Yes | Yes | ||
| EpCAM/TROP-1 | Yes | Yes | Yes | Yes | ||
| EphA2 | Yes | Yes | Yes | Yes | ||
| ErbB2/Her2 | Yes | Yes | Yes | Yes | Yes | Yes |
| ErbB3/Her3 | Yes | Yes | Yes | Yes | ||
| Flt-3/Flk-2 | Yes | Yes | Yes | Yes | ||
| GD2 | ||||||
| Glypican 3 | Yes | Yes | Yes | |||
| IL-3 R alpha/CD123 | Yes | Yes | Yes | |||
| IL-13 R alpha 2 | Yes | Yes | Yes | |||
| Mesothelin | Yes | Yes | Yes | Yes | Yes | Yes |
| MICA | Yes | Yes | Yes | Yes | Yes | |
| MICB | Yes | Yes | Yes | Yes | ||
| MICL/CLEC12A | Yes | |||||
| MUC-1 | Yes | Yes | Yes | |||
| NCAM-1/CD56 | Yes | Yes | Yes | Yes | Yes | |
| NKG2D | Yes | Yes | ||||
| PD-1 | Yes | Yes | Yes | Yes | ||
| PD-L1/B7-h1 | Yes | Yes | Yes | Yes | Yes | |
| PSMA/FOLH1 | Yes | Yes | Yes | |||
| ROBO1 | Yes | Yes | Yes | |||
| ROR1 | Yes | Yes | Yes | Yes | ||
| Siglec-2/CD22 | Yes | Yes | Yes | Yes | ||
| Siglec-3/CD33 | Yes | Yes | Yes | |||
| TIM-3 | Yes | Yes | Yes | Yes | Yes | |
| TROP-2 | Yes | Yes | Yes | |||
| WT1 | Yes |
Literature
- GMP Cytokines and Growth Factors for Therapeutic Manufacturing Brochure
- Accelerating Cell Therapy Discovery & Development With Non-Viral Gene Engineering Article
- Enhance Your TIL Expansion with ExCellerate T Cell Media Application Note
- Key Considerations for Cytokine Supplier Selection for Cell Therapies Article
- On the Cutting Edge of Immuno-Oncology with Simple Western and Single-Cell Western Scientific Review
- CAR T Cell Research Brochure
- CD4+ T Cell Subsets Brochure
- T Cell-Based Therapies eBook
- Current and Emerging Immune Checkpoint Targets for Immuno-Oncology Research eBook
- T Cell Subsets Poster
- Cell Markers Guide for Human Immune Cell Characterization Poster
- Immune Cell Isolation and Culture Brochure
- Immune Cell Therapy Workflow Poster
- Profiling Immune Cell Populations in the Tumor Microenvironment
- High Sensitivity Immunoassays for Detecting IFN-gamma in Cytokine Release Syndrome
Website Resources
- Immune Cell Culture
- Streamlining Transition to GMP
- Immune Cell Markers Interactive Tool at R&D Systems
- Flow Cytometry Panel Builder Tool
- Immunology Interactive Pathways at R&D Systems
- Immune Cell Differentiation and Characterization Protocols at R&D Systems
- Immunoassays for Immune Response Profiling
- Cell Expansion and Characterization Instruments
- Cell and Gene Therapy Response Profiling Instruments
On-Demand Webinars and Videos
- Cell and Gene Therapy Symposium: Streamlining Scale-Up and Scale-Out
- The Next Generation of AAV Characterization Tools
- Cell and Gene Therapy Symposium: Challenges of Analyzing ATMPs
- Cell and Gene Therapy Symposium: Will Non-Viral Gene Delivery Be the Future of Genetic Ex Vivo Modification?
- Analytical Solutions for Cell and Gene Therapy: Advancing Your AAV and CAR T-cell Development with Maurice and MFI
- Streamlining Cell Therapy IND Submission: Leveraging Raw Material Supplier Partnerships
- Advances in Cell and Gene Therapy Analytics: How to use Maurice to Characterize AAVs and LNPs
Effective immunotherapies can provide anti-tumor activity with the expression of chimeric antigen receptors (CAR) in T cells. CARs contain an extracellular tumor antigen-binding region (e.g. an scFv) and an intracellular region with at least one stimulatory domain. CAR T cells become activated following interactions with their cognate tumor cell-specific antigens. Alternatively, T cells can be engineered to express intact T cell receptors (TCR) with the desired antigen specificity. Generation of T cells with CARs or alternative TCRs is accomplished by genomic engineering with transposons (e.g. TcBuster), viral transduction, or nuclease systems such as CRISPR-Cas9. More complex genome engineering may additionally downregulate checkpoint molecules that would otherwise transmit immunosuppressive signaling.
T cell-based immunotherapy may induce anti-tumor responses and limit immune tolerance at the same time. Selection of the optimum T cell type(s) for T cell immunotherapy also contributes to the anti-tumor efficacy of adoptive cell therapies. CD4+ helper T cells, CD8+ cytotoxic T cells, central memory cells, and T memory stem cells exhibit distinct effector and memory functions as well as persistence in vivo. Cell therapies are classified as Advanced Therapy Medical Products (ATMPs) which are significantly more complex than purified pharmaceutical or biologic molecules. As living cellular treatments, ATMPs require extensive characterization to assure efficacy and patient safety.
*Luminex is a registered trademark of Luminex Corporation
*Alexa Fluor is a registered trademark of Molecular Probes Inc.