Recombinant Human IL-2 Heat Stable Agonist Protein, CF

Name: Recombinant Human IL-2 Heat Stable Agonist Protein, CF
Brand: R&D Systems
SKU: BT-002HS
Price: 143 USD
Availability: InStock

R&D Systems | Catalog # BT-002HS

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Product Documents

AI Modified Protein with thermostable synthetic core and enhanced beta subunit engagement, Analyzed by SEC-MALS

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Product Specifications
Scientific Data
Preparation & Storage
Calculators
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Simple to Use- IL-2 Heat Stable protein was evolved to both increase IL-2 beta receptor affinity and to also decrease IL-2 alpha receptor affinity for improved T cell and Tumor Infiltrating Lymphocyte (TIL) cell expansion compared to conventional IL-2 proteins.
Improved Stability- Enhanced stability facilitated by a synthetic core engineered to strengthen the protein's structure, enabling it to withstand high temperatures and extended culture durations.
Superior Capabilities- Higher bioactivity at 37 ^oC compared to standard IL-2.

Source

E. coli

Accession Number

Engineered based on P60568.1

Applications

Bioactivity

View all IL-2 Proteins and Enzymes »

Engineered using artificial intelligence (AI)
Redesigned synthetic protein core that confers thermostability for long-term experiments and storage capability
Unique receptor binding regions directed to CD122 with limited CD25 engagement
- Learn more in this Application Note

Product Specifications

Source

E. coli-derived human IL-2 protein
Proprietary

Purity

>95%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

SDS-PAGE

7 kDa, under reducing conditions.

Activity

Measured in a cell proliferation assay using NK-92 human natural killer lymphoma cells.
The ED₅₀ for this effect is 0.0150-0.150 ng/mL.

Scientific Data Images for Recombinant Human IL-2 Heat Stable Agonist Protein, CF

Recombinant Human IL‑2 Heat Stable Agonist AI Modified Protein SEC-MALS.

Recombinant Human IL‑2 Heat Stable Agonist AI Modified Protein (Catalog # BT-002HS) has a molecular weight (MW) of 12.6 kDa as analyzed by SEC-MALS, suggesting that this protein is a homodimer.

Heat Stable IL-2 retains activity for 59 days at 37 °C

Proliferation bioassay in NK92 cells after 59 days in media at 37 °C. Higher stability allows for less dosing throughout extended T cell cultures as well as early media prep to minimize culture inconsistencies from degrading cytokines.

Proliferation bioassay in NK92 cells after 10 min at 95 °C

After 10 min at 95°C (Control IL-2 C145S was kept at 4°C), only the heat stable IL-2 (Cat # BT-002HS) maintained bioactivity similar to the control IL-2, whereas the IL-2 Aldesleukin protein lost >14-fold activity.

Proliferation bioassay in NK92 cells after 10 days at 37°C

After 10 days at 10 ng/mL in the cell culture medium at 37°C (Control IL-2 C145S was kept at 4°C), only the heat stable IL-2 (Cat # BT-002HS) maintained bioactivity similar to the control IL-2, whereas the IL-2 Aldesleukin protein lost >3-fold activity.

Recombinant Human IL‑2 Heat Stable Agonist Protein Bioactivity.

Recombinant Human IL 2 Heat Stable Agonist (Catalog # BT-002HS) stimulates proliferation of NK-92 human natural killer lymphoma cells. The ED₅₀ for this effect is 0.0150‑0.150 ng/mL.

Recombinant Human IL‑2 Heat Stable Agonist Protein SDS-PAGE.

2 μg/lane of Recombinant Human IL‑2 Heat Stable Agonist Protein (Catalog # BT-002HS) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 7 kDa.

IL-2 Heat Stable Agonist Protein maintains exponential T cell growth through longer timepoints.

Heat Stable IL-2 (BT-002HS) Provides consistent CD4+/CD8+ T Cell Proliferation in Side-by-Side Testing with the Standard IL-2 Protein. CD4+ and CD8+ T cells were activated with anti-CD3-, anti-CD28- conjugated beads, and grown for 14 days in media containing 10 ng/mL Animal-free Recombinant Human IL-2 (R&D Systems, Catalog # BT-002-AFL) as the standard IL-2 protein or the AI modified Heat Stable IL-2. IL-2 conditions were refreshed every 2-3 days. The average fold expansion of the cells was determined every 2-3 days and showed that the Heat Stable IL-2 protein performs equally compared to the standard IL-2 protein.

Primary T Cells Expanded with Standard or Heat Stable IL-2 Display Similar Phenotypes.

CD4+ and CD8+ T cells were activated with anti-CD3-, anti-CD28- conjugated beads, and grown for 14 days in media containing 10 ng/mL Animal-free Recombinant Human IL-2 (R&D Systems, Catalog # BT-002-AFL) as the standard IL-2 protein or Heat Stable IL-2 (Cat # BT-002HS). IL-2 conditions were refreshed every 2-3 days. After 21 days, the phenotypes of the cells were compared by flow cytometry using a PE-Cy7-conjugated mouse anti-human CD45RA monoclonal antibody and a Brilliant Violet 711 conjugated mouse anti-human CCR7 monoclonal antibody to determine the number of CD4+ and CD8+ naive (CD45RA+ CCR7+), central memory (CD45RA- CCR7+), terminal effector memory (CD45RA+ CCR7- ), and effector memory (CD45RA- CCR7- ) T cells. The percentages of each of these cell populations were found to be similar whether the cells were expanded with the standard or Heat Stable IL-2 protein.

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Formulation, Preparation, and Storage

BT-002HS

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute the 10 μg size at 100 μg/mL in PBS. Reconstitute all other sizes at 500 μg/mL in PBS.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 12 months, 2 to 8 °C under sterile conditions after reconstitution. 12 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Background: IL-2

Predictive analytics are increasingly effective in solving challenges associated with developing protein reagents for cell culture applications, including thermal and pH instability. We have improved the stability of IL-2 by generating a synthetic structural core while maintaining efficiency in engagement and activation of IL-2 R beta / gamma _c receptors. The enhanced IL-2 displays improved heat stability enabling it to retain bioactivity while withstanding elevated temperatures and extended culture durations. Recombinant IL-2 is a bioactive protein for use in cell culture applications, often used for the expansion of T cells. IL-2 is expressed by CD4+ and CD8+ T cells, gamma delta T cells, B cells, dendritic cells, and eosinophils (1-3). rhIL-2 is also used for the expansion of NK cells, early differentiated T cells and effector memory Treg cells for adoptive cell transfer cancer immunotherapy (4,5). rhIL-2 Heat Stable agonist is verified for activity using an NK92 proliferation assay as well as T Cell culture applications comparing to the commonly used mutant aldesleukin IL-2 version.

References

Ma, A. et al. (2006) Annu. Rev. Immunol. 24:657.
Gaffen, S.L. and K.D. Liu (2004) Cytokine 28:109.
Taniguchi, T. et al. (1983) Nature 302:305.
Koehl, U. et al. (2015) Oncoimmunology. 5:e1115178.
Chamucero-Millares, J.A. et al. (2021) Cellular Immunol. 360:104257.

Long Name

Interleukin 2

Alternate Names

Aldesleukin, IL2, Proleukin, TCGF

Entrez Gene IDs

3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit)