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Part Number: EVEN-T
$300.00
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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.
NanoSparkTM 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.
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).
Need help getting started? Want to benefit most from your T cell activation and expansion? Contact the Nanotein team today to learn more.
Soluble, magnetic bead-free, protein-based, and ready-to-use.
Proprietary CD4+:CD8+ shifting technology with excellent total T cell expansion.
Employs anti-CD3/CD28 antibodies and a proprietary additive to modulate the CD4+:CD8+ ratio in a dose-dependent manner.
| 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) |
Consult the data sheets and supporting documentation for usage instructions and additional details, or explore the recommended protocols, which include suggested catalog numbers for cytokine supplements and media.
EVEN-T Cell Data Sheet 2024 (IL-7 & IL-15)
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.
Lot-specific EVEN-T CoA will be shipped with the product. Contact customer service if additional copies are required.
Carefully read product SDS before use.
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.
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.
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.
Increasing amounts of EVEN-T shifts the CD4+ :CD8+ ratio towards CD4+s with media-dependent differences in functional windows. NanoSparkTM EVEN-T Soluble T-Cell Activator was added to CD3+ negatively selected human peripheral blood T cells cultured in CellGenix’s TCM, Lonza’s T-Vivo, or FujiFilm’s Prime-XV serum-free media supplemented with IL-7 and IL-15 and compared to NanoSparkTM STEM-T Soluble T-Cell Activator. Cells were expanded for 10 days and analyzed on a flow cytometer on days 0, 7, and 10. Cells were labeled with CD4+ , CD8+ , CD45RA, CCR7, and CD95 fluorescent antibodies (Schmueck-Henneresse et al 2017). Expansion with NanoSparkTM EVEN-T Soluble T-Cell Activator modulates the expansion to shift the expanded T cell profile towards CD4+ T cells in a dose-dependent manner, effectively allowing the user to select a CD4+ :CD8+ T cell ratio at the outset of expansion.
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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