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Recombinant Human IL-2 GMP Protein, CF

Name: Recombinant Human IL-2 GMP Protein, CF
Brand: R&D Systems
SKU: BT-002-GMP
Price: 499 USD
Availability: InStock

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

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Citations (3)

Product Documents

Animal-Free. Contains the Cysteine to Serine substitution found in Proleukin® (aldesleukin)

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

Key Product Details

IL-2 Manufactured in Bio-Techne's new GMP facility
Lot-to-lot consistency
Stringent guidelines for patient safety
Scalability necessary to support successful therapeutics
Learn more about manufacturing in our new GMP facility
Test it in your process! Request a sample of GMP IL-2
A DMF is on file for this protein

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

<0.01 EU per 1 μg of the protein 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 Recombinant Human IL-2 GMP Protein, CF

Recombinant Human IL-2 GMP Protein Bioactivity.

GMP-grade Recombinant Human IL-2 (Catalog # BT-002-GMP) as measured in a cell proliferation assay using 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 GMP IL-2.

Equivalent Bioactivity of GMP and Animal-Free grades of Recombinant Human IL-2.

Equivalent bioactivity of GMP (Catalog # BT-002-GMP) and Animal-Free (BT-002-AFL) grades of Recombinant Human IL-2 as measured in a cell proliferation assay using CTLL-2 mouse cytotoxic T cells (orange and green, respectively).

Recombinant Human IL-2 GMP Protein SDS-PAGE.

2 μg/lane of Recombinant Human IL-2 GMP Protein (Catalog # BT-002-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 13 kDa.

Recombinant Human IL-2 GMP Protein Measured by Quantikine ELISA.

Two samples of GMP-grade human IL-2 (Catalog number BT-002-GMP, green circles) were interpolated with the human IL-2 Quantikine ELISA (D2050) standard curve (black circles) using 4PL logistic regression analysis. The human IL-2 Quantikine ELISA has an assay range of 31.2-2,000 pg/mL.

Recombinant Human IL-2 GMP Protein Measured by ELLA.

Thirteen samples of GMP-grade human IL-2 (Catalog number BT-002-GMP, green circles) were interpolated with the Simple Plex Human IL-2 cartridge (SPCKB-PS-000295) standard curve (black circles) using 4PL logistic regression analysis. The Simple Plex IL-2 cartridge has an assay range of 0.54-2,050 pg/mL.

Recombinant Human IL-2 GMP Protein Purity Analysis by Size Separation.

Three independent lots of Recombinant Human IL-2 GMP Protein (Catalog # BT-002-GMP) were analyzed by Maurice CE-SDS PLUS (IS is an Internal Standard). A gel-like representation of the purity analysis data (inset) can be obtained from the Lane View feature in Compass software for iCE. Profiles from the three runs were superimposed, showing excellent manufacturing consistency.

Recombinant Human IL-2 GMP Protein Purity Analysis by Charge Variation.

Three independent lots of Recombinant Human IL-2 GMP Protein (Catalog # BT-002-GMP) were analyzed by Maurice icIEF using native fluorescence detection (Mkr 5.85 and 9.99 are pI Markers). Profiles from the three runs were superimposed, showing excellent manufacturing consistency.

Expansion and viability of PBMCs.

Donor PBMCs were grown using ExCellerate NK Cell Expansion Media (Catalog # CCM037) supplemented with IL-2 (Catalog # BT-002-GMP), IL-12 (Catalog # 10018-IL), IL-15 (Catalog # BT-015-GMP), IL-18 (Catalog # BT-018-GMP), and IL-21 (Catalog # BT-021-GMP) on plates coated with anti-human NKp46 (Catalog # MAB1850). PBMCs were plated to contain 1.8e4 NK cells as determined by flow cytometry on day 0. On day 5, 0.1e6 total cells were taken to seed a 6 well plate. Viability, expansion, and NK purity were analyzed on day 9, with n=5 +/- SEM. A: Total expansion of CD56+ CD3- cells between day 0 and day 9. B: PBMCs grown in ExCellerate NK Cell Expansion Media supplemented with cytokines exhibit viability greater than 80% at day 9.

NK ExCellerate media preferentially expands CD56+ NK cells.

Donor PBMCs were grown using ExCellerate NK Cell Expansion Media (Catalog # CCM037) supplemented with IL-2 (Catalog # BT-002-GMP), IL-12 (Catalog # 10018-IL), IL-15 (Catalog # BT-015-GMP), IL-18 (Catalog # BT-018-GMP), and IL-21 (Catalog # BT-021-GMP) on plates coated with anti-human NKp46 (Catalog # MAB1850). PBMCs were plated to contain 1.8e4 NK cells as determined by flow cytometry on day 0. On day 5, 0.1e6 total cells were taken to seed a 6 well plate. Viability, expansion, and NK purity were analyzed on day 9, with n=5 +/- SEM for day 9; n=1 for day 0. A: Purity of NK cells at day 9. B: NK ExCellerate media preferentially expands CD56+ NK cells. Percentage of CD56+ cells in culture rose from less than 10% in both donors to more than 60% by day 9.

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Formulation, Preparation, and Storage

Lyophilized: BT-002-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in Sodium Acetate with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in sterile deionized water.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Liquid: BT-002-GMP/LQ

Formulation	Supplied as a 0.2 μm filtered solution in Sodium Acetate.
Shipping	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. A minimum of 12 months when stored at ≤ -65 °C as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 °C under sterile conditions after opening. 3 months, ≤ -20 °C under sterile conditions after opening.

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

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

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.
Mosmann, T.R. et al. (1987) J. Immunol. 138:1813.
Liparoto, S.F. et al. (2002) Biochemistry 41:2543.
Wang, X. et al. (2005) Science 310:1159.
Bodnar, A. et al. (2008) Immunol. Lett. 116:117.
Jaleco, S. et al. (2003) J. Immunol. 171:61.
Malek, T.R. (2003) J. Leukoc. Biol. 74:961.
Laurence, A. et al. (2007) Immunity 26:371.
Kryczek, I. et al. (2007) J. Immunol. 178:6730.
Afzali, B. et al. (2007) Clin. Exp. Immunol. 148:32.
Fehervari, Z. et al. (2006) Trends Immunol. 27:109.
Xue, D. et al. (2021) Antib Ther. 4:123.
Wolfarth, A.A. et al. (2022) Immune Netw. 22:e5.
Koehl, U. et al. (2015) Oncoimmunology. 5:e1115178.
Marsman, C. et al. (2022) Front. Immunol. 13:815449.
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 Recombinant Human IL-2 GMP Protein, CF

Download Product Datasheets: BT-002-GMP (lyophilized)

Download Product Datasheets: BT-002-GMP/LQ (liquid)

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.

Download SDS BT-002-GMP (lyophilized)

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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 Recombinant Human IL-2 GMP Protein, CF

Proleukin is a registered trademark of Clinigen Holdings Limited. 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