Recombinant Human IL-2, Animal-Free Protein

Newer Version Available: BT-002-AFL
NEW
Also available, GMP IL-2 Protein, Catalog # 202-GMP! Animal-free and manufactured under GMP guidelines for therapeutic manufacturing applications
Catalog # Availability Size / Price Qty
AFL202
AFL202-050
AFL202-010
Equivalent Bioactivity of GMP, Animal-Free, and RUO grades of Recombinant Human IL-2
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Citations (2)
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Recombinant Human IL-2, Animal-Free Protein Summary

Animal Free Proteins
Animal-free proteins are particularly important for researchers concerned with experimental variables caused by trace animal components or mammalian pathogens. Our products generated under animal-free conditions share the same biological activities as those produced using our standard laboratory techniques.

Product Specifications

Purity
>97%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<0.01 EU per 1 μg of the protein by the LAL method.
Activity
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 0.05-0.25 ng/mL.

The specific activity of recombinant human IL-2 is >5.0 x 106 IU/mg, which is calibrated against the human IL-2 WHO International Standard (NIBSC code: 86/500).
Source
E. coli-derived human IL-2 protein
Ala21-Thr153, with an N-terminal Met
Produced using non-animal reagents in an animal-free laboratory.
Accession #
N-terminal Sequence
Analysis
Met
Predicted Molecular Mass
15 kDa

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AFL202

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

AFL202

Formulation Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA with Trehalose.
Reconstitution Reconstitute at 0.2 mg/mL in sterile 100 mM Acetic Acid.  Alternatively, reconstitute at 0.1 mg/mL in sterile deionized water and use within 24 hours, store at 2 to 8 °C.
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 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Animal Free Proteins
Animal-free proteins are particularly important for researchers concerned with experimental variables caused by trace animal components or mammalian pathogens. Our products generated under animal-free conditions share the same biological activities as those produced using our standard laboratory techniques.

Scientific Data

Bioactivity View Larger

Equivalent bioactivity of GMP (202-GMP), Animal-Free (Catalog # AFL202) and RUO (202-IL/CF) grades of Recombinant Human IL-2 as measured in a cell proliferation assay CTLL 2 mouse cytotoxic T cells (orange, green, red, respectively).

Reconstitution Calculator

Reconstitution Calculator

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Background: IL-2

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.

References
  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) Antibody Therapeutics. 4(2): 123-133.
  15. Wolfarth, A.A. et al. (2022) Immune Netw. 22(1): e5.
  16. Koehl, U. et al. (2015) Oncoimmunology. 5(4).
  17. Marsman, C. et al. (2022) Front. In Immunol. 13(815449).
  18. Chamucero-Millares, J.A. et al. (2021) Cellular Immunology. 360(104257).
Long Name
Interleukin 2
Entrez Gene IDs
3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit)
Alternate Names
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

Manufacturing Specifications

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.  

 Production

  • All molecular biology procedures use animal-free media and dedicated labware.
  • Dedicated fermentors are utilized in committed animal-free areas.

Purification

  • Protein purification columns are animal-free.
  • Bulk proteins are filtered using animal-free filters.
  • Purified proteins are stored in animal-free containers in a dedicated cold storage room.

    Quality Assurance

    • Low Endotoxin Level.
    • No impairment of biological activity.
    • High quality product obtained under stringent conditions.
    • For ex vivo research or bioproduction, additional documentation can be provided.

    Please read our complete Animal-Free Statement

    Citations for Recombinant Human IL-2, Animal-Free Protein

    R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

    2 Citations: Showing 1 - 2
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    1. Eighty-eight variants highlight the role of T cell regulation and airway remodeling in asthma pathogenesis
      Authors: TA Olafsdotti, F Theodors, K Bjarnadott, US Bjornsdott, AB Agustsdott, OA Stefansson, EV Ivarsdotti, JK Sigurdsson, S Benonisdot, GI Eyjolfsson, D Gislason, T Gislason, S Guðmundsdó, A Gylfason, BV Halldorsso, GH Halldorsso, T Juliusdott, AM Kristinsdo, D Ludviksdot, BR Ludviksson, G Masson, K Norland, PT Onundarson, I Olafsson, O Sigurdardo, L Stefansdot, G Sveinbjorn, V Tragante, DF Gudbjartss, G Thorleifss, P Sulem, U Thorsteins, GL Norddahl, I Jonsdottir, K Stefansson
      Nat Commun, 2020;11(1):393.
      Species: Human
      Sample Types: Whole Cells
      Applications: Cell Culture
    2. IL-15 promotes activation and expansion of CD8+ T cells in HIV-1 infection
      Authors: Souheil-Antoine Younes
      J Clin Invest, 2016;126(7):2745-2756.
      Species: Human
      Sample Types: Whole Cells
      Applications: Bioassay

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