Recombinant Human IL-2 Heat Stable Agonist Protein, CF

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.

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.

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 (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.

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.

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.

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.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human IL-2 Heat Stable Agonist Protein, CF Summary

• Additional Information: AI Modified Protein with thermostable synthetic core and enhanced beta subunit engagement
• Details of Functionality: 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.
• Source: E. coli-derived human IL-2 protein
  Proprietary
• Accession #: Engineered based on P60568.1
• Protein/Peptide Type: Recombinant Proteins
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• SDS-PAGE: 7 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Reconstitution Instructions: Reconstitute the 10 μg size at 100 μg/mL in PBS. Reconstitute all other sizes at 500 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human IL-2 Heat Stable Agonist Protein, CF

• Aldesleukin
• IL2
• IL-2
• IL-2lymphokine
• interleukin 2
• interleukin-2
• involved in regulation of T-cell clonal expansion
• Proleukin
• T cell growth factor
• T-cell growth factor
• TCGF

Background

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.

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

Bioinformatics

• Uniprot:
  • Engineered based on P60568.1()