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Natural Killer (NK) Cells for Cell Therapy

Natural killer cells have the potential to address common limitations of T cell-based immune cell therapies. NK cell biology offers promising advantages for both therapeutic activity and manufacturing simplification, leading to improved delivery of effective therapy to patients.

  • Reduced risk of cell therapy failure due to antigen escape. NK cells act by several mechanisms besides antigen specific CAR targeting. Their redirected cell lysis and antibody-directed cellular cytotoxicity (ADCC) functions serve as antigen-independent means to kill mutant target tumor cells.
  • Improved scalability and speed of therapy delivery. NK cells are amenable to developing allogeneic therapies based on universal cell banks. These banks can supply therapeutic cells much more rapidly than autologous processes which require patient cells to produce each treatment.
NK cell genetically engineered for immune cell therapy

As Your Program Advances Towards the Clinic, We Will Grow With You

Adherent cells in culture 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
  • Simplify comparability testing for raw material changes
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.

NK Cell Isolation

Begin your immunotherapy workflow on a strong footing. Accurate and reproducible isolation of NK cell populations will lay the groundwork for a successful cell therapy program. After cell isolation, confirm that your procedure is effective with thorough cell phenotype analysis.

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

NK Cell Selection

Antibody specificity is the basis for accurate and reproducible cell selection. Choosing the right antibodies will enable you to reliably isolate high purity NK cell populations.

View Custom Antibody Services for 

  • Recombinant antibody engineering and GMP antibodies
  • Monoclonal panel development
  • Specialty conjugation

 

    induced pluripotent cells in culture for differentiation into NK cells

    NK Cells From Induced Pluripotent Stem Cells

    Differentiation of iPSCs into NK cells facilitates the generation of universal cell banks to treat multiple patients with reduced cost and more rapid development.

    NK Cell Culture and Expansion

    Generating enough NK cells to support clinical trials requires a consistent and scalable expansion protocol. To boost culture consistency, QC your media preparations with precise quantitation of cytokines and growth factors. Analyze your expanded cultures for cell number, viability, and functionality.

    NK cell
    NK Excellerate Bottle Teaser

    NK Cell Culture Media

    Support your NK cell culture for translational studies with serum-free and animal component-free media such as ExCellerate Human NK Cell Expansion Media. This media enables the robust expansion of CD56+ cells from total human PBMCs or purified CD3-depleted PBMCs.

    Profile GMP and secreted cytokines

    GMP cytokines and growth factors for NK cell therapy manufacturing

    Animal-Free and GMP Cytokines & Growth Factors

    Ease your transition to clinical NK cell manufacturing by supplementing your cultures with Animal-Free and GMP Proteins that have equivalent bioactivity to research-grade cytokines. Then quantitate the cytokines in your media with precise and repeatable ELISA kits.

    functional antibody binding to NK cell to trigger activation and expansion

    Agonist Antibodies

    Enhance your NK cell expansion and activation steps with functional antibodies that activate specific signal transduction pathways. Target multiple NK cell receptors including KIR2DL4, NKG2D, NKp30, NKp44, and NKp46. Simplify activation with the CellXVivo Human NK Cell Expansion Kit.

    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 feeder-free NK cell expansion, delivering true platform, process, and product continuity for your NK 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.

    NK Cell Engineering

    Enhance the performance of your NK cell-based therapy and realize the potential of gene engineering. Partner with our experts for customized number of cells/vial, validation testing, datasheet preparation, and batch record documentation. We will work with you to make sure we engineer the right solution for your goals.

    Watch the video to see how TcBuster delivers your genes of interest.

    TcBuster for Non-Viral Gene Delivery

    Accomplish your NK cell engineering more easily by using non-viral gene delivery. TcBuster will revolutionize your ability to engineer the optimum phenotype for your cell therapy.

    • Reduced cost of genetic modification
    • Rapid cell engineering with manufacturing controls
    • Large cargo size integration
    • Flexibility to deliver multiple genes in one operation
    • Not subject to supply shortage of lentivirus vectors

    NK Cell Engineering With Viral Vectors

    Vector characterization

    Process efficiency improvements with viral transduction enhancers

    Automated AAV viral titer assays developed in partnership with PROGEN

    Learn More About Vector Characterization

    AAV and lentiviral viral vector manufacturing solutions
    analysis of NK cell phenotype after cell engineering

    Confirming Successful Cell Engineering

    Once you have engineered your NK cells, verify that their phenotype reflects the gene editing you intended.

    flow cytometry analysis of NK cells after transduction with human CD19 CAR construct

    Human NK cells were enriched from PBMCs and either (A) transduced or (B) not transduced with a human CD19 CAR. Cells were stained with Human CD56 PE-conjugated Antibody and Recombinant Human CD19 Protein, Atto 647N Conjugate.

    Fluorokine App Note

    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.

    BioInsights Webinar Foreground

    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.

    NK Cell Characterization

    Final product characterization of your NK cells should cover all critical quality attributes (CQAs) to ensure your product is as safe as possible for in vivo administration. Using orthogonal assays to measure unrelated parameters will enable thorough analysis of the cell surface, secretory profile, NK cell cytolytic activity, and residual contaminants.

    characterization of NK cells for immune cell therapy
    phenotype of NK cell for immune cell therapy

    NK Cell Phenotype

    Perform a detailed phenotypic characterization of your NK cells to determine if they express the right surface receptors for the cell subset and activation status you need. In addition, measure population heterogeneity and the absence of residual feeder cells from culture expansion.

    analysis of NK cells with the FlowX Human NK Cell Phenotyping Kit

    NK cell phenotypic analysis with the FlowX Human NK Cell Phenotyping Kit. NK cells were expanded from PBMCs in ExCellerate Human NK Cell Expansion Media with plate-bound anti-human NKp46 supplemented with recombinant human IL-2, IL-12, IL-18, and IL-21 for 13 days. Cells were analyzed with antibody conjugates included in the kit: NKp30  Alexa Fluor 488, CD16 PerCP, CD56 Alexa Fluor™ 647, NKG2D Alexa  Fluor 700, NKp46 PE, and CD3. Gating was based off live cells (using the Live-or-Dye 405/545 dead cell exclusion dye), and quadrants were set using isotype controls (not shown).

    immunoassay detection of effector molecules released by activated NK cells

    NK Cell Function

    To make sure your cells are functional as well as phenotypically intact, assay their release of effector NK cell molecules including IFN-gamma, Granzymes, and Perforin. In addition, directly measure the ability of your cells to lyse target cells in co-culture.

    analysis of NK cell cytotoxicity with the FlowX Human NK Cell Killing Flow Cytometry Kit

    CAR-NK cell cytotoxicity using the FlowX Human NK Cell Killing Flow Cytometry Kit. Human NK cells were enriched from PBMCs and transduced with a human CD19 CAR (CAR-NK). CAR-NKs were labeled with Janelia Fluor® 646, Daudi target cells or K562 target cells were labeled with MitoMark Green, and cells were co-cultured for 2 hours at the indicated Effector to Target ratios. Anti-LAMP-1/CD107a PE antibody was included in the culture during the 2 hour killing assay to measure degranulation. Cells were harvested and stained with anti-human CD56 APC and CD3 Alexa Fluor 405 antibodies. To measure target cell death, CD3-/CD56- Janelia Fluor 646- cells were analyzed for MitoMark Green.

    Watch the video to see how MFI distinguishes different types of particles in a cell culture sample.

    Contaminant Screening

    Make sure that your NK cell product is clean enough for in vivo administration, either in animal models or in clinical trials. Confirm that the levels of particulates, residual cell activation beads, and other impurities are low enough to meet your requirements.

    Watch the video for an overview of RNAscope in situ hybridization technology.

    Tissue Biopsy and NK Cell Localization

    Monitor the biodistribution of your NK cell therapy in vivo with single-cell resolution in the context of tissue morphology.

    • RNAscope and BaseScope assays for quantitative transgene mRNA analysis in intact tissues
    • Good Clinical Laboratory Practice (GCLP)-compliant RNAscope ISH analysis for clinical and preclinical studies
    • Professional Assay Services for tissue-based expression analysis support
    RNAscope Multiplex Fluorescent V2 Assay detection of NK cells in situ in cervical cancer

    Detection of NK cells in human cervical cancer using the RNAscope Multiplex Fluorescent V2 Assay. NK cells were detected in situ with specific marker probes for NCR1, CD56, XCL1, and CCL5. Read the scientific poster for more details.

    Luminex Intelliflex xMAP instrument with drawer closed

    In Vivo Host Monitoring

    Profile the host response to your NK cell therapy with the R&D Systems® Luminex® Assays. The Human XL Cytokine Performance panel profiles up to 46 targets simultaneously. Choose the format that meets your needs.

    Custom Services

    No cutting edge cell therapy should be delayed from reaching patients just because the right materials aren’t available. If you need specially designed reagents or assays for developing your NK 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.

    Luminex® is a Registered Trademark of Luminex Corporation.