GMP

ProPak™ Recombinant Human IL-2 GMP Protein

R&D Systems | Catalog # PPK-002-GMP

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Key Product Details

  • Closed-System Process In order to de-risk your therapy as much as possible, ProPak GMP cytokines are designed to integrate seamlessly into a closed-system process. The risk of contamination is significantly lowered as a result. Learn more about closed process reagents.
  • Weldable Tubing Each ProPak bag is outfitted with weldable tubing, so you can connect it directly to your media, then add both to your bioreactor of choice. Tubing is compatible with any bagged media for use in a 1 L bioreactor, such as the G-Rex.
  • Ready-to-Use Format Supplied in a 3mL frozen liquid format. Extensive quality testing guarantees that the liquid formulation is just as potent and stable as lyophilized equivalents, allowing you to safely take away the need for reconstitution steps.
  • Reduced Manual Touchpoints By removing both reconstituting and aliquoting steps, there is less hands-on technician time. This drastically reduces risks of error while streamlining your journey to the clinic.

See the ProPak User Manual to learn how to add ProPak GMP cytokines into your closed process. 

Source

E. coli

Accession Number

P60568.1

Applications

Bioactivity
View all IL-2 Proteins and Enzymes »

Product Specifications

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.

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 Level

<5.0 EU/mL by the LAL method.

N-terminal Sequence Analysis

Met-Ala21-Pro-Thr-Ser-Ser-Ser-Thr-Lys-Lys & Ala21-Pro-Thr-Ser-Ser-Ser-Thr-Lys-Lys-Thr

Predicted Molecular Mass

15.5 kDa

SDS-PAGE

13 kDa, under reducing conditions.

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 0.0300-0.250 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).

Host Cell Protein

<0.500 ng per μg of protein when tested by ELISA.

Mycoplasma

Negative for Mycoplasma.

Host Cell DNA

<0.00150 ng per µg of protein when tested by PCR.

Scientific Data Images for ProPak™ Recombinant Human IL-2 GMP Protein

Lot to Lot Consistency

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™ cytokines are successfully recovered into 1L media bags.

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™ cytokines support T cell growth

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™ cytokines support lentiviral transduced T cell expansion

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.

Formulation, Preparation, and Storage

PPK-002-GMP
Formulation Supplied as a 0.2 μm filtered solution in PBS, recombinant HSA, and Trehalose.
Reconstitution
Shipping This product is shipped on dry ice. Upon receipt, store immediately at the temperature recommended below.
Stability & 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.

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.

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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 delta 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) 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

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)

Gene Symbol

IL2

UniProt

P60568.1

Additional IL-2 Products

Product Documents for ProPak™ Recombinant Human IL-2 GMP Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

R&D Systems' quality focus includes:

  • Designed, manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
  • Documented and controlled manufacturing process
  • Control of documentation and process changes by QA
  • Personnel training programs
  • Raw material inspection and vendor qualification/monitoring program
  • Validated equipment, processes and test methods
  • Equipment calibration and maintenance schedules using a Regulatory Asset Manager
  • Facility/Utilities maintenance, contamination controls, safety and pest control programs
  • Material review process for variances
  • Robust product stability program following relevant ICH guidelines

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their application. Each product is provided with a lot-specific Certificate of Analysis that contains the product's specifications and test results. Quality control testing may include, but is not limited to:

  • N-terminal amino acid analysis
  • SDS-PAGE purity analysis
  • Molecular weight analysis via mass spectrometry
  • Endotoxin assessment per USP <85> and Ph. Eur. 2.6.14 guidelines
  • Bioassay analysis
  • Microbial testing per USP <71> and Ph. Eur. 2.6.1 guidelines
  • Host cell protein assessment
  • Host cell DNA assessment
  • Mycoplasma assessment

Additional testing and documentation requested by the customer can be arranged at an additional cost. 

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis and St. Paul, Minnesota USA.

R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.


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.

    Please read our complete Animal-Free Statement

    Product Specific Notices for ProPak™ Recombinant Human IL-2 GMP Protein

    Full terms and conditions of sale can be found online in the Protein Sciences Segment T&Cs at: Terms & Conditions.

    For preclinical, or clinical ex vivo use

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