ProPak™ Recombinant Human IL-2 GMP Protein, CF

ProPak™ GMP-grade Recombinant Human IL-2 (PPK-002-GMP) stimulates proliferation of CTLL‑2 mouse cytotoxic T cells. The ED50 for this effect is 0.0300-0.250 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of ProPak™ GMP IL-2.

ProPak™ IL-2 (PPK-002-GMP-010) was added to Xeno-free Human T cell media in a bag. In parallel, lyophilized IL-2 (BT-002-GMP) was added to Xeno-free Human T cell media in a bottle (CCM038-GMP-1L). After cytokine addition, the concentration of IL-2 (SPCKB-PS-000295) was determined by the Ella automated immunoassay system. Data is the average of 4 independent experiments with error bars ±SD.

ProPak IL-2 (PPK-002-GMP-010) was added to Xeno-free Human T cell media with 5% human AB serum. In parallel, lyophilized IL-2 (BT-002-GMP) was added to Xeno-free Human T cell media in a bottle (CCM038-GMP-1L). T cells from 3 donors were activated and expanded in G-Rex bioreactors for 7 days and assessed for cells/cm2 and cell viability. Data is the average of 4 independent experiments with 3 donors per experiment, error bars ±SD.

ProPak™ IL-2 (PPK-002-GMP-010) was added to Xeno-free Human T cell media with 5% human AB serum. In parallel, lyophilized IL-2 (BT-002-GMP) was added to Xeno-free Human T cell media in a bottle (CCM038-GMP-1L). T cells from 3 donors were activated for 1 day then transduced with eF1a-CD19-DHFR-eGFP lentivirus. Cells were expanded in a G-Rex® bioreactor for a total of 7 days, then assessed for fold expansion and cell viability. Data is the average of 3 donors, error bars ±SD.

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

ProPak™ Recombinant Human IL-2 GMP Protein, CF Summary

• Details of Functionality: Measured in a cell proliferation assay using CTLL‑2 mouse cytotoxic T cells. Gearing, A.J.H. and C.B. Bird (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 295.

  The ED₅₀ for this effect is 0.0300-0.250 ng/mL.

  The specific activity of recombinant human IL-2 is >5.00 x 10⁶ IU/mg, which is calibrated against an internal reference standard value assigned against the human IL-2 WHO International Standard (NIBSC code: 86/500).

• Source: E. coli-derived human IL-2 protein
  Ala21-Thr153 (Cys145Ser), with and without an N-terminal Met
  Produced using non-animal reagents in an animal-free laboratory.
  Manufactured and tested under cGMP guidelines.
• Accession #: P60568.1
• N-terminal Sequence: Met-Ala21-Pro-Thr-Ser-Ser-Ser-Thr-Lys-Lys & Ala21-Pro-Thr-Ser-Ser-Ser-Thr-Lys-Lys-Thr
• Protein/Peptide Type: GMP Recombinant Proteins
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 15521 ± 5 Da, and a second 15390 ± 5 Da product may be present
• Endotoxin Note: <5.0 EU/mL by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 15.5 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 13 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  
  A minimum of 6 months when stored between -14 °C and -40 °C. Can be stored up to 2 weeks at 2-8 °C. Refer to lot specific COA for the Use by Date.
• Buffer: Supplied as a 0.2 μm filtered solution in PBS, recombinant HSA, and Trehalose.
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 15521 ± 5 Da, and a second 15390 ± 5 Da product may be present

Notes

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For preclinical, or clinical ex vivo use
Not for in vivo use


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

Alternate Names for ProPak™ Recombinant Human IL-2 GMP 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

Recombinant Interleukin-2 (IL-2) is expressed in E. coli and has been engineered to contain the serine for cysteine substitution found in Proleukin® (aldesleukin). Recombinant IL-2 is widely used in cell culture for the expansion of T cells. IL-2 is expressed by CD4+ and CD8+ T cells, gamma δ T cells, B cells, dendritic cells, and eosinophils (1-3). Mature human IL-2 shares 56% and 66% amino acid (aa) sequence identity with mouse and rat IL-2, respectively. Human and mouse IL-2 exhibit cross-species activity (4). The receptor for IL-2 consists of three subunits that are present on the cell surface in varying preformed complexes (5-7). The 55 kDa IL-2 R alpha is specific for IL-2 and binds with low affinity. The 75 kDa IL-2 R beta , which is also a component of the IL-15 receptor, binds IL-2 with intermediate affinity. The 64 kDa common gamma chain gamma c/IL-2 R gamma , which is shared with the receptors for IL-4, -7, -9, -15, and -21, does not independently interact with IL-2. Upon ligand binding, signal transduction is performed by both IL-2 R beta and gamma c. IL-2 is best known for its autocrine and paracrine activity on T cells. It drives resting T cells to proliferate and induces IL-2 and IL-2 R alpha synthesis (1, 2). It contributes to T cell homeostasis by promoting the Fas-induced death of naïve CD4+ T cells but not activated CD4+ memory lymphocytes (8). IL-2 plays a central role in the expansion and maintenance of regulatory T cells, although it inhibits the development of Th17 polarized cells (9-11). Thus, IL-2 may be a key cytokine in the natural suppression of autoimmunity (12, 13). IL-2 expression and concentration can have either immunostimulatory effects at high doses or immunosuppressive effects at low doses due to its preferential binding to different receptor subunits expressed by various immune cell types. This has led to the generation of recombinant IL-2 variants aimed at modifying IL-2 receptor binding for increased antitumor efficacy (14, 15). These variants are typically used in combination with immune checkpoint inhibitors instead of as a monotherapy (14). IL-2 can be genetically engineered to express in NK cells for CAR T cell therapies, and in combination with other cytokines like IL-15, can increase cell viability and proliferation (16). In addition to adoptive cell transfer and checkpoint blockade inhibitors, cancer vaccines that boost immune responses have been combined with IL-2 treatment with promising results in recent studies (15). In cell culture, IL-2 is a frequently used cytokine for the proliferation, differentiation, and increased antibody secretion of B cells as they transform into plasma cells in vitro (17). IL-2 is also a classically used cytokine for the expansion of NK cells, early differentiated T cells and effector memory Treg cells for adoptive cell transfer cancer immunotherapy (16, 18). GMP IL-2 is a commonly used supplement for the expansion of these cell types for cellular therapies.

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. Mosmann, T.R. et al. (1987) J. Immunol. 138:1813.
5. Liparoto, S.F. et al. (2002) Biochemistry 41:2543.
6. Wang, X. et al. (2005) Science 310:1159.
7. Bodnar, A. et al. (2008) Immunol. Lett. 116:117.
8. Jaleco, S. et al. (2003) J. Immunol. 171:61.
9. Malek, T.R. (2003) J. Leukoc. Biol. 74:961.
10. Laurence, A. et al. (2007) Immunity 26:371.
11. Kryczek, I. et al. (2007) J. Immunol. 178:6730.
12. Afzali, B. et al. (2007) Clin. Exp. Immunol. 148:32.
13. Fehervari, Z. et al. (2006) Trends Immunol. 27:109.
14. Xue, D. et al. (2021) Antib Ther. 4:123.
15. Wolfarth, A.A. et al. (2022) Immune Netw. 22:e5.
16. Koehl, U. et al. (2015) Oncoimmunology. 5:e1115178.
17. Marsman, C. et al. (2022) Front. Immunol. 13:815449.
18. Chamucero-Millares, J.A.et al. (2021) Cell Immunol. 360:104257

Bioinformatics

• Uniprot:
  • P60568.1()