Recombinant Human IL-18/IL-1F4 GMP Protein, CF GMP

Name: Recombinant Human IL-18/IL-1F4 GMP Protein, CF
Brand: R&D Systems
SKU: BT-018-GMP

R&D Systems, part of Bio-Techne | Catalog # BT-018-GMP

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

Key Product Details

IL-18 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-18

Source

E. coli

Accession #

Q14116.1

Conjugate

Unconjugated

Applications

Bioactivity

View all IL-18/IL-1F4 Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human IL-18/IL-1F4 protein
Tyr37-Asp193
Produced using non-animal reagents in an animal-free laboratory.Manufactured and tested under cGMP guidlines.

Purity

>97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 18205 Da ± 5 Da.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Tyr37-Phe-Gly-Lys-Leu-Glu-Ser-Lys-Leu-Ser

Predicted Molecular Mass

18 kDa

SDS-PAGE

18 kDa, under reducing conditions.

Activity

Measured by its ability to induce IFN-gamma secretion by KG-1 human acute myelogenous leukemia cells in the presence of TNF-alpha.
The ED₅₀ for this effect is 1.25-15.0 ng/mL.

The specific activity of GMP Recombinant Human IL-18/IL-1F4 is > 5.00 x 10⁶ units/mg, which is calibrated against the human IL-18 WHO Standard (NIBSC code: 03/200).

Host Cell Protein

<0.200 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-18/IL-1F4 GMP Protein, CF

Recombinant Human IL-18/IL-1F4 GMP Protein Bioactivity.

The bioactivity of Recombinant Human IL-18/IL-1F4 GMP Protein (Catalog # BT-018-GMP) was measured by its ability to induce IFN-gamma secretion by KG-1 human acute myelogenous leukemia cells in the presence of TNF-alpha. Three independent lots were tested for bioactivity and plotted on the same graph to show lot-to-lot consistency of GMP IL-18/IL-1F4 protein.

Equivalent Bioactivity of GMP and Animal-Free grades of Recombinant Human IL‑18/IL‑1F4.

Equivalent bioactivity of GMP (Catalog # BT-018-GMP) and Animal-Free (BT-018-AFL) grades of Recombinant Human IL-18/IL-1F4 as measured by its ability to induce IFN-gamma secretion by KG-1 human acute myelogenous leukemia cells in the presence of TNF-alpha (orange and green, respectively).

Recombinant Human IL‑18/IL‑1F4 GMP Protein SDS-PAGE.

2 μg/lane of Recombinant Human IL‑18/IL‑1F4 GMP Protein (Catalog # BT-018-GMP) was resolved with SDS-PAGE under reducing (R)and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing a band at 18 kDa under reducing conditions.

NK cells grown from PBMCs using a cytokine cocktail containing GMP IL-18 show robust expansion and high viability at day 9.

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 cells in PBMC culture are highly expansive when grown in ExCellerate NK Cell Expansion Media containing GMP cytokines.

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

BT-018-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS and TCEP with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in sterile 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.

Background: IL-18/IL-1F4

Interleukin-18 (IL-18) is a proinflammatory cytokine in the IL-1 family that exerts distinct immune effects depending on the local cytokine environment. It is expressed as a 24 kDa precursor by endothelial and epithelial cells, keratinocytes, gamma delta T cells, and phagocytes. The precursor is activated intracellularly by Caspase-1 mediated proteolysis to release the 17 kDa mature cytokine. The precursor can also be released by necrotic cells for extracellular cleavage by multiple proteases. IL-18 activation is induced by infection or tissue damage and contributes to disease pathology in chronic inflammation (1-3). IL-18 binds to the widely expressed IL-18 R alpha which recruits IL-18 R beta to form the signaling receptor complex (4, 5). Its bioactivity is negatively regulated by interactions with IL-18 binding proteins and virally encoded IL-18BP homologs (6). In the presence of IL-12 or IL-15, IL-18 enhances anti-viral Th1 immune responses by inducing IFN-gamma production and the cytolytic activity of CD8⁺ T cells and NK cells (7, 8). In the absence of IL-12 or IL-15, however, IL-18 promotes production of the Th2 cytokines IL-4 and IL-13 by CD4⁺ T cells and basophils (9, 10). In the presence of IL-1 beta or IL-23, IL-18 induces the antigen-independent production of IL-17 by gamma delta T cells and CD4⁺ T cells (11). IL-18 also promotes myeloid dendritic cell maturation and triggers neutrophil respiratory burst (12, 13). In cancer, IL-18 exhibits diverse activities including enhancing anti-tumor immunity, inhibiting or promoting angiogenesis, and promoting tumor cell metastasis (14). Mature human IL-18 shares approximately 63% amino acid sequence identity with mouse and rat IL-18 (15). Alternative splicing in human ovarian cancer generates an isoform that is resistant to Caspase-1 activation (16). A cell surface form can be expressed on M-CSF induced macrophages and released in response to bacterial endotoxin (17).

References

Dinarello, C.A. et al. (2013) Front. Immunol. 4:289.
Smith, D.E. (2011) J. Leukoc. Biol. 89:383.
Gu, Y. et al. (1997) Science 275:206.
Torigoe, K. et al. (1997) J. Biol. Chem. 272:25737.
Cheung, H. et al. (2005) J. Immunol. 174:5351.
Novick, D. et al. (1999) Immunity 10:127.
Fehniger, T.A. et al. (1999) J. Immunol. 162:4511.
Yoshimoto, T. et al. (1998) J. Immunol. 161:3400.
Yoshimoto, T. et al. (2000) Nat. Immunol. 1:132.
Kroeger, K.M. et al. (2009) J. Leukoc. Biol. 86:769.
Lalor, S.J. et al. (2011) J. Immunol. 186:5738.
Li, J. et al. (2004) Cell. Immunol. 227:103.
Elbim, C. et al. (2005) Clin. Diagn. Lab. Immunol. 12:436.
Fabbi, M. et al. (2015) J. Leukoc. Biol. 97:665.
Ushio, S. et al. (1996) J. Immunol. 156:4274.
Gaggero, A. et al. (2004) Oncogene 23:7552.
Bellora, F. et al. (2012) Eur. J. Immunol. 42:1618.

Long Name

Interleukin 18

Alternate Names

IGIF, IL-1F4, IL-1g, IL18

Entrez Gene IDs

3606 (Human); 16173 (Mouse); 29197 (Rat); 397057 (Porcine); 574151 (Primate)

Gene Symbol

IL18

UniProt

Q14116.1

Additional IL-18/IL-1F4 Products

Product Documents for Recombinant Human IL-18/IL-1F4 GMP Protein, CF

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COA

SDS

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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:

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, mass spectrometry, 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 <71>
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-18/IL-1F4 GMP Protein, CF

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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