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Immune Cell Therapy NK Cell

Immune Cell Therapy Antibodies

Immune cell therapy is rapidly emerging as an important alternative for cancer treatment, as well as treatment of autoimmune disease. Also known as adoptive cell therapy or cellular immunotherapy, it supercharges an individual’s immune cells to fight disease.

There are several ways to harness the power of the immune system for cellular immunotherapy:

  • Harvest and expand tumor infiltrating lymphocytes (TILs), endogenous T cells and Natural Killer (NK) cells in the tumor microenvironment (TME).
  • Genetically engineer T cells with a unique tumor-antigen specific TCR.
  • Knockout inhibitory receptors like PD-1, CTLA-4 or NKG2A in isolated T cells or NK cells.
  • Insert chimeric antigen receptors (CAR)s, which include an antigen-recognition domain linked directly to a costimulatory domain, into T cells or NK cells.
    • CAR T cells bypass the requirement for antigen presentation by MHC I.
    • CAR NK cells impart antigen specificity to these potently cytotoxic cells.

Antibodies are critical tools for each step of the cellular immunotherapy workflow. They can be used to isolate, activate, and characterize both the cellular input and the final immune cell therapy product. Whether your goal is to isolate and expand TILs or engineer CAR T cells or CAR NK cells, Bio-Techne has validated antibodies and other quality reagents for each stage of the manufacturing process. Learn more about how Bio-Techne reagents and instruments can streamline and enhance your T cell manufacturing or NK cell therapy workflow.

Custom GMP Antibody Services

  • Take your immune cell therapy to the clinic with our custom GMP antibodies for ex vivo use.
  • Develop an antibody from scratch to your exact specifications.
  • Convert an existing research-grade antibody into a recombinant GMP antibody.
Custom GMP Antibody graphic

Antibodies for Immune Cell Activation

In vitro activation and expansion are required to insert CAR constructs for CAR T cells and CAR NK cells and increase the number of cells for infusion.  Antibodies with agonist activity are commonly used to activate both T cells and NK cells in vitro. We utilize our vast selection of in-house manufactured proteins and our bioassay expertise to develop the most appropriate cellular assay for testing antibody activity, including assays for cellular proliferation, chemotaxis, cytokine secretion, adhesion, and enzyme neutralization. Additionally, we are always expanding our assay repertoire to ensure the highest specificity based on the latest research publications.

*Functional antibodies for CD2 and FcγR III (CD16) found at Novus Biologicals.

Cell Type

Marker

T cell

CD3, CD28, 4-1BB/TNFSRF9/CD137

NK cell

CD2*, FcγR III (CD16)*, NKp46/NCR1

Cytokine Blocking Antibodies

Different CD4+ T helper (TH) cell subsets serve different immune functions. TH1 cells primarily have pro-inflammatory anti-tumor and anti-viral activity. while TH2 cells can promote a pro-tumor, anti-inflammatory environment. TH17 cells have been reported to contribute to both tumor progression and tumor elimination. Depending on the desired final T cell therapy product, isolated TH cells can be polarized in vitro using anti-cytokine blocking antibodies in combination with recombinant growth factors and cytokines

T Cell Subset

Anti-Cytokine Antibody

TH1 Cells

IL-4

TH2 Cells

IFN-γ, IL-12

TH17 Cells

IFN-γ, IL-4

Learn more about our commitment to quality and validation of blocking and neutralization antibodies from R&D Systems. 

T Cell Stimulation Induced by Human CD3 epsilon Antibody

Functional Antibodies Induce Cytokine Secretion

(Left) T Cell Stimulation Induced by Human CD3 epsilon Antibody. Mouse Anti-Human CD3e Monoclonal Antibody (Catalog # MAB100R) induces stimulation in human T cells in a dose-dependent manner, as measured by Resazurin (Catalog # AR002), when immobilized at 1-10 µg/mL (100 µL/well). (Right) Human CD28 Antibody Enhances IL-2 Secretion in Jurkat Cells. Human CD28 Monoclonal Antibody enhances IL-2 secretion in the Jurkat human acute T cell leukemia cell line stimulated with 10 ng/mL phorbol myristate acetate (PMA) and 0.5 µM calcium ionophore, in a dose-dependent manner, as measured using the Quantikine Human IL-2 ELISA Kit (Catalog # D2050). The ED50 for this effect is typically 0.2-0.6 µg/mL.

Additional Reagents for Immune Cell Activation & Expansion

Enrichment of CD4+ T Cells from Human Peripheral Blood Mononuclear Cells Using the MagCellect CD4+ T Cell Isolation Kit

MagCellect Kits for Immune Cell Isolation

  • Antibody-based kits for the isolation or enrichment of specific immune cells from a heterogeneous cell population.
  • Verify the purity of your isolated population using our rigorously tested antibodies validated for flow cytometry.
Cloudz™ Cell Activation Kit

Cloudz™ Cell Activation Kits

  • Ready-made kits to expand total T cells, Regulatory T cells (Tregs), or NK cells using dissolvable microspheres.
  • Customize your activation and expansion with Screening CloudzTM kit.
GMP Certified Proteins graphic

GMP Cytokines and Growth Factors

  • Industry-leading GMP cytokines and growth factors manufactured under GMP guidelines
  • Choose GMP cytokines with bioactivity that is benchmarked against research-grade cytokines to minimize process disruption during the transition.
Conjugated Antibody Flow Cytometry Cell

Antibodies for Immune Cell Characterization

During the manufacturing process, it is often critical to isolate specific populations of immune cells from peripheral blood mononuclear cells (PBMCs) or the patient's tumor. Ensure you are isolating the relevant populations of immune cells, using Bio-Techne’s rigorously tested, highly validated antibodies for flow cytometry. Learn more about our commitment to reproducibility and antibody validation.

After isolation and expansion, antibodies are also required to assess the critical quality attributes (CQAs) of your immune cell therapy preparation, like purity and potency. Have confidence in your results using conjugated primary antibodies from Bio-Techne, backed by our 100% Guarantee and validated for flow cytometry.

Purity of immune cell therapy preparation refers to the frequency of cells that are desired cell type (e.g., T cell or NK cell). For CAR T cell or CAR NK cell therapy, purity also refers to the frequency of cells within the preparation that express chimeric antigen receptor. 

T Cells  
Cell Populations Markers
Hematopoietic Lineage CD45
All T cells CD3
CD4+ T cells CD4
CD8+ T cells CD8
γ/δ T cells γ/δ TCR*
iNKT cells Vα24Jα18 TCR*
Naïve T cells L-selectin/CD62L, CCR7
T regulatory cells (Tregs) CD25/IL-2Rα

*Antibodies for γ/δ TCR  and Vα24Jα18 TCR are available from Novus Biologicals.  

NK Cells  
Cell Populations Markers
Hematopoietic Lineage CD45
NK cells NCAM-1/CD56
NK cells for ADCC Fc Gamma R III (CD16)
Pro-inflammatory NK cells CD161

 

 The majority of NK cells found in peripheral blood can be characterized based on their expression of CD16 and CD56. NK cells are traditionally defined as CD16+CD56dim or CD16dimCD56+. 

 

 

Find more NK Cell Markers below.

Fluorescent tagged proteins illustration

Fluorescent Proteins for CAR Detection

To evaluate the purity of your CAR cell preparation, use conjugated fluorescent proteins to save time and avoid high background staining associated with secondary antibodies. Bio-Techne offers fluorescent-labeled CD19, BCMA, CD22/Siglec-2 and more! Inquire about other targets with Custom Protein Services.

Potency of immune cell therapy includes in vitro determination of direct cytotoxic activity, as well as quality and quantity of cytokines produced. The definition of potency can be expanded to include a complete understanding of the phenotype of your immune cell preparation including differentiation status and expression of inhibitory receptors.

Select Antibodies for T cell Characterization

Pro-Inflammatory Cytokines
IFN-γ
IL-2
TNF-a
Perforin
Granzyme B
Anti-Inflammatory Cytokines
IL-10
TGF-β
IL-35
Galectin-1

*Visit Novus Biologicals to find flow-validated antibodies that detect these cell markers.

Key Antibodies for NK Cell Characterization

To phenotype your NK cell therapy, Bio-Techne offers highly validated monoclonal and recombinant monoclonal antibodies for NK cell surface proteins including the Natural Cytotoxicity Receptor (NCR) and Killer-Immunoglobulin-Like Receptors (KIR) families.

 FlowX  Human NK Cell Phenotyping Flow Cyometry Kit (Catalog # FMC033)

Bio-Techne's multi-color flow cytometry kits have the reagents you need to streamline your flow cytometry panel design, including staining buffers and isotype controls.

Marker

Clone

Fluorochrome

CD3

UCHT1*

Alexa Fluor 405

NCAM1/CD56

2524C

Alexa Fluor 647

NKp30/NCR3

210845*

Alexa Fluor 488

NKG2D/CD314

149810*

Alexa Fluor 700

NKp46/NCR1

195314*

PE

Fc Gamma R III (CD16)

245536

PerCP

*CD3 (clone UCHT1), NKG2D/CD314 (clone 149810), NKp30/NCR3 (clone 210845), and NKp46/NCR1 (clone 195314) were independently validated by HLDA and used to designate CD nomenclature of target.

 
NK cell phenotype by flow cytometry graph

NK cell phenotype by flow cytometry. PBMCs were activated with Cloudz Human NK Cell Expansion Kit (Catalog # CLD004) and stained with the following antibodies from the FlowX Human NK Cell Phenotyping Flow Cytometry Kit (Catalog # FMC033) : Alexa Fluor 405®-conjugated anti-CD3 epsilon (clone UCHT1) Alexa Fluor® 647-conjugated anti-NCAM-1/CD56 (clone 2524C), Alexa Fluor® 488-conjugated anti-NKp30/NCR3 (clone 210845) , Alexa Fluor® 700-conjugated anti-NKG2D/CD314 (clone 149810) , PE-conjugated  anti-NKp46/NCR1 (clone 195314) , and  PerCP-conjugated anti-Fc Gamma R III (CD16) (clone 245536).

Additional Tools for Immune Cell Phenotyping

NK cell foreground image

R&D Systems Cell Marker Tool

There are many cell types in the immune system that can be engineered and optimized for new cellular therapies, including cytotoxic T cells, NK Cells, Regulatory T cells, and γδ T cells.  Explore markers for these immune cells, and more, with the  Interactive Cell Marker Tool.

Flow cytometry

Novus Flow Cytometry Panel Builder

Streamline your flow cytometry panel design process using the Panel Builder from Novus Biologicals. Select your cytometer configuration then choose compatible fluorochromes from a selection of highly validated conjugated primary antibodies. 

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 immune cell therapy solutions, delivering true platform, process, and product continuity for your immune 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.