GMP

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

R&D Systems | Catalog # BT-018-GMP

R&D Systems
100% Guarantee

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 Number

Q14116.1

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.

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.

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.

=
÷

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

  1. Dinarello, C.A. et al. (2013) Front. Immunol. 4:289.
  2. Smith, D.E. (2011) J. Leukoc. Biol. 89:383.
  3. Gu, Y. et al. (1997) Science 275:206.
  4. Torigoe, K. et al. (1997) J. Biol. Chem. 272:25737.
  5. Cheung, H. et al. (2005) J. Immunol. 174:5351.
  6. Novick, D. et al. (1999) Immunity 10:127.
  7. Fehniger, T.A. et al. (1999) J. Immunol. 162:4511.
  8. Yoshimoto, T. et al. (1998) J. Immunol. 161:3400.
  9. Yoshimoto, T. et al. (2000) Nat. Immunol. 1:132.
  10. Kroeger, K.M. et al. (2009) J. Leukoc. Biol. 86:769.
  11. Lalor, S.J. et al. (2011) J. Immunol. 186:5738.
  12. Li, J. et al. (2004) Cell. Immunol. 227:103.
  13. Elbim, C. et al. (2005) Clin. Diagn. Lab. Immunol. 12:436.
  14. Fabbi, M. et al. (2015) J. Leukoc. Biol. 97:665.
  15. Ushio, S. et al. (1996) J. Immunol. 156:4274.
  16. Gaggero, A. et al. (2004) Oncogene 23:7552.
  17. 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

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 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

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

    There are currently no reviews for this product. Be the first to review Recombinant Human IL-18/IL-1F4 GMP Protein, CF and earn rewards!

    Have you used Recombinant Human IL-18/IL-1F4 GMP Protein, CF?

    Submit a review and receive an Amazon gift card!

    $25/€18/£15/$25CAN/¥2500 Yen for a review with an image

    $10/€7/£6/$10CAN/¥1110 Yen for a review without an image

    Submit a review
    Amazon Gift Card

    FAQs

    No product specific FAQs exist for this product.

    View all FAQs for Proteins and Enzymes