Recombinant Human IL-15 GMP Protein, CF

GMP-grade Recombinant Human IL-15 (Catalog # BT-015-GMP) stimulates cell proliferation in the MO7e human megakaryocytic leukemic cell line. The ED50for this effect is 0.300-2.60 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP IL-15.

2 μg/lane of GMP-grade Recombinant Human IL-15 (Catalog # BT-015-GMP) was resolved with SDS-PAGE under reducing (R) conditions and visualized by Coomassie® Blue staining, showing a single band at 9 kDa.

Two samples of GMP-grade human IL-15 (Catalog number BT-015-GMP, green circles) were interpolated with the human IL-15 Quantikine ELISA (D1500) standard curve (black circles) using 4PL regression analysis. The human IL-15 Quantikine ELISA has an assay range of 3.9-250 pg/mL.

Thirteen samples of GMP-grade human IL-15 (Catalog number BT-015-GMP, green circles) were interpolated with the Simple Plex Human IL-15 cartridge (SPCKB-PS-000500) standard curve (black circles) using 4PL logistic regression analysis. The Simple Plex IL-15 cartridge has an assay range of 0.2-1,950 pg/mL.

Three independent lots of Recombinant Human IL-15 GMP Protein (Catalog # BT-015-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.

Three independent lots of Recombinant Human IL-15 GMP Protein (Catalog # BT-015-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.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human IL-15 GMP Protein, CF Summary

• Additional Information: Animal-Free
• Details of Functionality: Measured in a cell proliferation assay using MO7e human megakaryocytic leukemic cells. The ED 50 for this effect is 0.300-2.60 ng/mL. The specific activity of recombinant human IL-15 is >2.00 x 10 8 units/mg, which is calibrated against the human IL-15 reference standard (NIBSC code: 95/554).
• Source: E. coli-derived human IL-15 protein Asn49-Ser162 Produced using non-animal reagents in an animal-free laboratory. Manufactured and tested under cGMP guidelines.
• Accession #: P40933.1
• N-terminal Sequence: Asn49-Trp-Val-Asn-Val-Ile-Ser-Asp-Leu-Lys
• Protein/Peptide Type: GMP Recombinant Proteins
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 12761 Da ± 5 Da
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 13 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 9 kDa, under reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  A minimum of 12 months when stored at ≤ -20 degreesC as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 3 months, ≤ -20 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 12761 Da ± 5 Da
• Reconstitution Instructions: Reconstitute at 100-500 μg/mL in PBS.

Notes

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Alternate Names for Recombinant Human IL-15 GMP Protein, CF

• IL15
• IL-15
• IL-15MGC9721
• interleukin 15
• interleukin-15

Background

Interleukin 15 (IL-15) is a widely expressed 14 kDa cytokine that is structurally and functionally related to IL-2 and plays an important role in many immunological diseases (1, 2). Mature human IL-15 shares 70% amino acid sequence identity with mouse and rat IL-15. Alternative splicing generates isoforms of IL-15 with either a long or short signal peptide (LSP or SSP), and the SSP isoform is retained intracellularly (3). IL-15 binds with high affinity to IL-15 R alpha (4). It binds with lower affinity to a complex of IL-2 R beta and the common gamma chain ( gamma c) which are also subunits of the IL-2 receptor complex (5). IL-15 associates with IL-15 R alpha in the endoplasmic reticulum, and this complex is expressed on the cell surface (6). The dominant mechanism of IL-15 action is known as transpresentation in which IL-15 and IL-15 R alpha are coordinately expressed on the surface of one cell and interact with complexes of IL-2 R beta / gamma c on adjacent cells (7). This enables cells to respond to IL-15 even if they do not express IL-15 R alpha (6). In human and mouse, soluble IL-15-binding forms of IL-15 R alpha can be generated by proteolytic shedding and bind up nearly all the IL-15 in circulation (8-10). Soluble IL-15 R alpha functions as an inhibitor that limits IL-15 action (4, 9). Ligation of membrane-associated IL-15/IL-15 R alpha complexes also induces reverse signaling that promotes activation of the IL-15/IL-15 R alpha expressing cells (11). IL-15 induces or enhances the differentiation, maintenance, or activation of multiple T cell subsets including NK, NKT, Th17, Treg, and CD8+ memory cells (12 - 16). An important component of these functions is the ability of IL-15 to induce dendritic cell differentiation and inflammatory activation (11, 14). IL-15 exhibits anti-tumor activity independent of its actions on NK cells or CD8+ T cells (17). It also inhibits the deposition of lipid in adipocytes, and its circulating levels are decreased in obesity (18). Immunotherapy treatment with recombinant IL-15 has the advantage of not stimulating Treg cells like IL-2 does but has the drawback of associated toxicity at higher doses. This has led to increased investigation on mitigating IL-15 toxicity and combination immunotherapy approaches using immune checkpoint inhibitors (19, 20). Preclinical and early clinical studies have shown the potential of also using IL-15 in combination with cancer vaccines to improve their anti-tumor response (20). IL-15 can also be used for the preconditioning of CAR T cells or for engineering cells to express IL-15 in vivo. Adoptive cell transfer of NK cells engineered to express CD19 and IL-15 were well tolerated in patients with CD19-positive cancers (20). IL-15 can be used in combination with other cytokines like IL-21 to increase the efficiency of NK cell expansion and maturation in stem cell culture protocols (21). The combination of IL-15 with IL-7 also promotes expansion of early-differentiated CD8+ T cells in culture with the added benefit of decreasing Treg cell generation, unlike IL-2, for adoptive cell transfer in cancer immunotherapy (22). GMP IL-7 and GMP IL-15 are commonly used in combination for ex vivo expansion of T cells for cellular therapies.

1. De Sabatino, A. et al. (2011) Cytokine Growth Factor Rev. 22:19.
2. Grabstein, K. et al. (1994) Science 264:965.
3. Tagaya, Y. et al. (1997) Proc. Natl. Acad. Sci. USA 94:14444.
4. Giri, J.G. et al. (1995) EMBO J. 14:3654.
5. Giri, J. et al. (1994) EMBO J. 13:2822.
6. Dubois, S. et al. (2002) Immunity 17:537.
7. Castillo, E.F. and K.S. Schluns (2012) Cytokine 59:479.
8. Budagian, V. et al. (2004) J. Biol. Chem. 279:40368.
9. Mortier, E. et al. (2004) J. Immunol. 173:1681.
10. Bergamaschi, C. et al. (2012) Blood 120:e1.
11. Budagian, V. et al. (2004) J. Biol. Chem. 279:42192.
12. Mortier, E. et al. (2003) J. Exp. Med. 205:1213.
13. Gordy, L.E. et al. (2011) J. Immunol. 187:6335.
14. Harris, K.M. (2011) J. Leukoc. Biol. 90:727.
15. Xia, J. et al. (2010) Clin. Immunol. 134:130.
16. Schluns, K.S. et al. (2002) J. Immunol. 168:4827.
17. Davies, E. et al. (2010) J. Leukoc. Biol. 88:529.
18. Barra, N.G. et al. (2010) Obesity 18:1601.
19. Xue, D. et al. (2021) Antib Ther. 4:123.
20. Wolfarth, A.A. et al. (2022) Immune Netw. 22:e5.
21. Oberoi, P. et al. (2020) Cells. 9:811.
22. Chamucero-Millares, J.A. et al. (2021) Cellular Immunol. 360:104257.

Bioinformatics

• Uniprot:
  • P40933.1()