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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.

T Cell

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

adherent cells in culgture supported by extracellular matrix

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
gowned cleanroom operator carrying cell culture vessel

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
gowned cleanroom operator carrying cell culture vessel

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.

cell surface receptors bound by antibodies for NK cell isolation
antibodies used for NK 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.

View Custom Antibody Services

  • Recombinant antibody engineering and GMP antibodies
  • Monoclonal panel development
  • Specialty conjugation
induced pluripotent cells in culture for differentiation into NK cells

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.

Browse iPSC Reagents

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.

T cell targeted by Cloudz for cell culture expansion
bottle of excellerate

T Cell Culture Media

Improve the consistency of your T cell cultures with serum-free and xeno-free media. These media enable robust T cell expansion in the presence of Cloudz Human T Cell Activation Kits, cytokines, or plate-bound antibodies.

GMP cytokines and growth factors for NK cell therapy manufacturing

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.

functional antibody binding to NK cell to trigger activation and expansion

Activating Antibodies

For T cell expansion and activation, utilize functional antibodies to activate specific signal transduction pathways. engage primary and costimulatory signals with CD3 and CD28 antibodies.

Cloudz™ for Cell Activation and Expansion

Cloudz™ Cell Activation Kits

Streamline and optimize your T cell expansion with the simple 1-2-3 protocol of Cloudz kits. Cloudz reagents are derivatized alginate-based hydrogel microspheres which rapidly dissolve with the addition of Release Buffer. No bead removal step is required. You can fine tune T cell activation and expansion with alternate Cloudz ligands which enable you to modulate specific receptors and signal transduction pathways.

T Cell Expansion and Activation Graphs

T cell expansion and activation using the GMP Cloudz™ Human T Cell Activation Kit. Primary human CD3+ cells were activated with Cloudz and cultured for 9 days in ExCellerate™ T Cell Expansion Media supplemented with 20 ng/mL GMP Human IL-2. (A) Cells were counted to determine fold expansion compared to the Day 0 seeding density (0.25 x 106 cells/mL). (B, C) After 48 hours of culture, cells were analyzed by flow cytometry using CD25-PE and CD69-Alexa Fluor® 488 antibody conjugates. (C) Analysis of a negative control culture performed without the addition of Cloudz T Cell microspheres.

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.

ScaleReady Logo

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.

T Cell Engineering

Realize the potential of gene engineering and enhance the performance of your T cell-based therapy. Cell engineering enables you to fine tune the effectiveness of your T cell therapy by introducing and deleting molecules that alter cell phenotype and function. Partner with our experts so we can engineer the right solution for your goals.

  • Introduce bispecific, tandem, or multiple chimeric antigen receptors (CARs)
  • Modulate activating and inhibitory receptors
  • Overexpress stimulatory cytokines
  • Secrete scFv for checkpoint inhibition
  • Express chemokine receptors for improved trafficking in vivo
non-viral NK cell engineering with TcBuster gene delivery system
TcBuster(TM) - A next-generation non-viral gene delivery system

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%.

CAR-T cell engineering with TcBuster

T Cell Engineering With Lentivirus

 

Vector characterization

Lentiviral vector titration with HIV-1 Gag p24 ELISAs and Reagents

Process efficiency improvements with viral transduction enhancers

Learn About Vector Characterization at ProteinSimple

AAV and lentiviral vector manufacturing solutions
Cell Engineering With TcBuster Graphic

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.

flow cytometry analysis of CAR-T cells with Recombinant Human CD19 Protein, Atto 488 Conjugate

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.

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).

characterization of NK cells for immune cell therapy
induced pluripotent cells in culture for differentiation into NK cells

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.

Analysis of T cells with the FlowX Human Th1 Cell Multi-Color Flow Cytometry Kit

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).

immunoassay detection of effector molecules released by activated NK cells

T Cell Secretory Profile

Evaluate the functionality of your cells by analyzing their secretory profile including key cytokines such as IFN-gamma, IL-2, and TNF-alpha. You can expect accurate and reliable results with immunoassays built with carefully tested, in-house antibodies, proteins, and specialized diluents.

correlation of Simple Plex and Quantikine HS ELISA for quantitating human IFN-gamma. multianalyte quantitation with the Human Immunotherapy Magnetic Luminex Performance Assay 24-plex Fixed Panel.

(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.

Micro-Flow Imaging: I See Particles

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 analysis of T cells and beads in a mixed population

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.

ACD RNAscope in situ Hybridization (ISH) Technology Overview

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
RNAscope Multiplex Fluorescent V2 Assay detection of CAR-T cells within tumor site

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.

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.

image of T Cell-Based Therapies eBook

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.

*Luminex is a registered trademark of Luminex Corporation

*Alexa Fluor is a registered trademark of Molecular Probes Inc.