NanoSpark™ EVEN-T
Soluble T Cell Activator

The NanoSparkTM EVEN-T Soluble T Cell Activator reagent kit allows the user to modulate the CD4+:CD8+ ratio in a dose-dependent manner during T cell expansion, including CAR-T cells.
  • First-in-Class CD4+:CD8+ ratio shifting technology
  • Soluble format
  • All protein-based
  • No magnetic components
  • Anti-CD3/CD28 based
  • GMP in production, available ~Q2/Q3 2024

Add NanoSparkTM EVEN-T Soluble T Cell Activator to your cart now!

Part Number: EVEN-T

$300.00

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Description

Product Highlights

The NanoSparkTM EVEN-T Soluble T Cell Activator is a culture-ready reagent for in vitro activation and expansion of human T cells. The nanoscale of the NanoSparkTM EVEN-T protein complex allows for sterile filtration and easy removal of the activating reagent via standard media exchange or simple centrifugal cell washing. The percentage of CD4+ T cells can be increased by adding increasing amounts of EVEN-T to the culture media, effectively modulating the CD4+:CD8+ ratio during the expansion step. The functional dose range is media-specific and can be tuned to your platform needs.

  • Convenient

    Soluble, magnetic bead-free, protein-based, and ready-to-use.

  • High Performance

    Proprietary CD4+:CD8+ shifting technology with excellent total T cell expansion.

  • Reliable

    Employs anti-CD3/CD28 antibodies and a proprietary additive to modulate the CD4+:CD8+ ratio in a dose-dependent manner.

What Is a T Cell Activator?

A T cell activator is a substance or molecule that stimulates the activation and proliferation of T cells, enabling the uptake of genetic constructs for CAR-T applications and encouraging rapid and robust expansion in in vitro culture.

In the body, CD8+ and CD4+ T cells require antigen-presenting cells (APCs) to initiate activation. This is a complex and costly process that is impractical to replicate ex vivo. Simplified reagent-based activation and expansion protocols using T cell activators are largely preferred.

Reagent-based activation is also ideally suited for CAR-T therapy development. This is especially true when large quantities are required for T cell immunotherapy research and manufacturing.

How Does the NanoSparkTM EVEN-T Soluble T Cell Activator Work?

The NanoSparkTM EVEN-T Soluble T Cell Activator contains self-assembling protein nanoparticles with functional antibodies bound to the surface through high-affinity non-covalent interactions. These anti-CD3 and anti-CD28 antibodies are arranged in a proprietary biophysical combination on the NanoSparkTM platform. EVEN-T is formulated with a safe, proprietary additive that enables shifting of the CD4+:CD8+ T cell ratio throughout the expansion.

The NanoSparkTM EVEN-T Soluble T Cell Activator activates resting human CD3+ T cells originating from purified T lymphocytes or patient peripheral blood mononuclear cells (PBMCs).

Nanotein vs. Other Products

NanoSpark(TM) EVEN-T is first in class – there are no other T cell activator products that allow researchers to control the CD4+:CD8+ T cell ratio during expansion. Competing products introduce clinical risks to the process by using synthetic ingredients that may contaminate T cell therapy if not painstakingly removed from the expanded culture.

The NanoSparkTM product line is modular and plug-and-play by design, using a proprietary self-assembly protein nanoparticle complex that is inherently soluble and can be filter sterilized.

Inquire About Nanotein’s T Cell Activator

Need help getting started? Want to benefit most from modulating the CD4+:CD8+ T cell ratio in your immunotherapy research? Contact the Nanotein team today to learn more.

Product Details

Product Line NanoSparkTM
Cell Type T cells
Sample Type PBMC, cell culture, T cell isolate
Reactivity Human
Application T cell activators, T cell manufacturing, immunotherapy
Grade Research grade
Quantity 1 mL
Contents Activator contents suspended in phosphate-buffered saline glycerol
Functional Testing ≥80% expected proliferation of T cells on day 7
Sterility Sterile 0.2 µm filtered post-aseptic production
Mycoplasma Negative
Storage Keep frozen at -80°C for long-term storage
Once thawed, store at 4°C protected from light for up to one month
Shipping Condition Frozen on dry ice
Product Weight 20 g (0.044 lbs)
Product Dimensions 13 x 13 x 38 mm (0.5 x 0.5 x 1.5 in)

Protocols

The NanoSparkTM EVEN-T Soluble T Cell Activator is used in vitro to induce activation, modulate CD4+/CD8+ T cell ratio, and CAR gene delivery uptake T cells. It is intended for ex vivo activation and expansion of CD3+ T Lymphocytes (human resting T cells) from T cell isolates or peripheral blood mononuclear cells (PBMCs). The activator should be stored at -80 °C long-term. Once thawed, store at 4 °C protected from light for up to one month.

The NanoSparkTM EVEN-T Soluble T Cell Activator is designed for use with a cytokine-supplemented T cell expansion medium. Suggested protocols have been developed for common cytokine-supplemented strategies. Optimization of the provided general protocol information may be necessary depending on your experimental objectives.

IL-2 

Interleukin 2 (IL-2) is a potent general T cell growth factor essential for the long-term proliferation of activated T cells.

IL-2 should be added to culture media at 20 ng/mL final concentration. Check back for detailed EVEN-T IL-2 protocol, but see analogous IL-2 STEM-T Protocol and Data Sheet for general guidelines.

IL-7/IL-15

Interleukin 7 (IL-7) is a cytokine required in early T cell development that promotes T cell survival by up-regulating the expression of anti-apoptotic genes. Interleukin 15 (IL-15) is an inflammatory cytokine that supports the homeostasis of naive CD8+ T cells. Combining IL-7 and IL-15 in culture promotes the enrichment of self-renewing T cells like TSCM.

IL-7 and IL-15 should be added to culture media at 10 ng/mL final concentration of each. For detailed information, please refer to the IL-7/IL-15 EVEN-T Protocol and Data Sheet.

EVEN-T has yet to be optimized in Wilson Wolf Sterile GRex6M cell culture vessels, so check back for a detailed protocol. Until then, please see IL-7/IL-15 GRex6M STEM-T Protocol and Data Sheet for general guidelines.

Tips for Best Results Using the NanoSparkTM EVEN-T Soluble T Cell Activator

  • Seed CD3+ T cells/PBMCs at 7 x 105 cells/cm2 in culture media.
  • Add 5-10 µL of NanoSparkTM EVEN-T Soluble T Cell Activator for every mL of cell suspension (e.g., 1,000 µL of Activator for 100 mL of cell suspension) based on the media used per Figure 1.
  • Incubate cells at 37 °C and 5% CO2 in a humidified incubator.
  • For cryopreserved cells, gently thaw cells and exchange them into culture media before starting.
  • For optimal cell expansion and maintenance, ensure the activator is in culture media (conditioned or fresh) for at least 72 hours (removal is unnecessary for expansion).
  • Be sure to add fresh cytokine-supplemented culture media every three days.

If using transduction (optional), wait 24-72 hours after adding NanoSparkTM EVEN-T Soluble T Cell Activator to the cell culture before adding the viral vector. If using electroporation, then electroporate 72 hours after activator addition.

Data & Research

Data Sheets

These protocols are recommended and include suggested catalog numbers of cytokine supplements and media to use.
View EVEN-T Cell Data Sheet 2024 (IL-7 & IL-15)

Certificate of Analysis (CoA) Template

Lot-specific EVEN-T CoA will be shipped with the product. Contact customer service if additional copies are required.
View Nanotein Example STEM-T CoA

Safety Data Sheet (SDS)

Carefully read product SDS before use.
View NanoSparkTM EVEN-T SDS

FAQ's

Quality & Safety

The underlying NanoSparkTM platform has undergone biocompatibility testing when formulated with antibodies specific to our T cell activator, STEM-T, which we consider a significant derisking step for the platform as a whole. The details are as follows:

The composition of the NanoSparkTM reagent is protein-based, and consequently naturally biodegradable, which provides a significant safety advantage over magnetic bead-based and synthetic products. A reputable medical device CRO was commissioned to complete the in vitro & in vivo ISO 10993 biocompatibility testing.  The tests were run at >20X the predicted possible carry-over amount, which resulted in no toxicity or adverse effects:

ISO 10993 

Biocompatibility Testing

Test Type of Test Result Product Tested
ISO 10993-5 Cytotoxicity In vitro No Toxicity/Adverse Effect NanoSpark STEM-T
ISO 10993-10 Sensitization In vivo: Guinea Pig No Toxicity/Adverse Effect NanoSpark STEM-T
ISO 10993-10 Irritation/Intracutaneous Reactivity In vivo: Rabbits No Toxicity/Adverse Effect NanoSpark STEM-T
ISO 10993-11 Acute Systemic Toxicity In vivo: Mice No Toxicity/Adverse Effect NanoSpark STEM-T
ISO 10993-11 Pyrogenicity In vivo: Rabbits No Toxicity/Adverse Effect NanoSpark STEM-T
ISO 10993-4 Hemocompatibility In vitro No Toxicity/Adverse Effect NanoSpark STEM-T
ISO 10993-3 Genotoxicity Ames In vitro No Toxicity/Adverse Effect NanoSpark STEM-T
ISO 10993-3 Genotoxicity Mouse Lymphoma Forward Mutation In vitro No Toxicity/Adverse Effect NanoSpark STEM-T

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