Recombinant Human IL-2 GMP Protein, CF GMP

Name: Recombinant Human IL-2 GMP Protein, CF
Brand: R&D Systems
SKU: BT-002-GMP

Catalog # BT-002-GMP | R&D Systems, Inc. a Bio-Techne Brand

Product Datasheet / COA / SDS

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

New! Bypass reconstitution steps by using a liquid formulation of GMP-grade Recombinant Human IL-2. Find out more here.

Setting the standard in quality research reagents

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

Source:	E. coli
Accession #:	P60568.1
Applications:	Bioactivity

View Full Technical Details for Recombinant Human IL-2 GMP Protein, CF

Technical Data
Citations / Reviews

Product Specifications for Recombinant Human IL-2 GMP Protein, CF

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 visualized with Silver Staining and 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	15.5 kDa
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 the human IL-2 WHO International Standard (NIBSC code: 86/500).

Scientific Data Examples 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.

Formulation, Preparation and Storage

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.

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.

Reconstitution Calculator

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 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.

Item	Value
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
Entrez Gene IDs	3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit)
Alternate Names	Aldesleukin, IL-2, IL-2lymphokine, IL2, Proleukin, T cell growth factor, T-cell growth factor, TCGF, interleukin 2, interleukin-2, involved in regulation of T-cell clonal expansion

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:

Manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented processes and QA control of documentation and process changes
Personnel training programs
Raw material testing and vendor qualification/monitoring
Fully validated equipment, processes and test methods
Equipment calibration schedules using a computerized calibration program
Facility maintenance, safety programs and pest control
Material review process for variances
Monitoring of stability over product shelf-life

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 research. The Certificate of Analysis provided contains the following lot specific information:

N-terminal amino acid analysis, SDS-PAGE analysis, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial testing according to USP
Host Cell Protein testing performed by ELISA
Mycoplasma testing by ribosomal RNA hybridization assay

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, 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 in a dedicated cold storage room.

Quality Assurance

Low Endotoxin Level.
No impairment of biological activity.
High quality product obtained under stringent conditions.

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.

END USER TERMS OF USE OF PRODUCT

The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and comply with all of the provisions of these End User Terms of Use of Product, you should contact your supplier of the product and make arrangements to return the product.

We suggest you print and retain a copy of these End User Terms of Use of Product for your records.

The End User is aware that R&D Systems, Inc. sells GMP products for preclinical or clinical ex vivo use and not for in vivo use. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.

R&D Systems, Inc. has the right, at its sole discretion, to modify, add or remove any terms or conditions of these End User Terms of Use without notice or liability to you. Any changes to these End User Terms of Use are effective immediately following the printing of such changes on this product insert. The most recent version of these End User Terms of Use of Product may be found at: RnDSystems.com/Legal.

You agree to review these End User Terms of Use of Product to ensure any subsequent use by you of R&D Systems' GMP Products following changes to these End User Terms of Use of Product constitutes your acceptance of all such changes.

TERMS AND CONDITIONS

The following limitation applies to R&D Systems' warranty and liability for damages: All products are warranted to meet R&D Systems' published specifications when used under normal laboratory conditions.

R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.

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Full details of R&D Systems' Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal.

Citations for Recombinant Human IL-2 GMP Protein, CF

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