ProPak™ Recombinant Human IL-7 GMP Protein, CF

ProPak GMP-grade Recombinant Human IL-7 (PPK-007-GMP) stimulates proliferation of PHA-activated human peripheral blood lymphocytes. The ED50 for this effect is 0.100-0.500 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of ProPak GMP IL‑7.

ProPak IL-7 (PPK-007-GMP) and ProPak IL-15 (PPK-015-GMP) were added to Xeno-free Human T cell media in a bag (CCM038-GMP-1B). In parallel, liquid IL-7 (BT-007-GMP-025/LQ) and IL-15 (BT-015-GMP-025/LQ) or lyophilized IL-7 (BT-007-GMP) and IL-15 (BT-015-GMP) were added to Xeno-free Human T cell media in a bottle (CCM038-GMP-1L). After cytokine addition, the concentration of IL-7 (SPCKB-CS-003676) and IL-15 (SPCKB-CS-003688) was determined by the Ella automated immunoassay system. Data is the average of 4 independent experiments with error bars ±SD.

ProPak IL-7 (PPK-007-GMP) and ProPak IL-15 (PPK-015-GMP) were compared liquid IL-7 (BT-007-GMP-025/LQ) and IL-15 (BT-015-GMP-025/LQ) or lyophilized IL-7 (BT-007-GMP) and IL-15 (BT-015-GMP). Each were added to Xeno-free Human T cell media in a bag (CCM038-GMP-1B) with 5% huAB Serum. T cells from 3 donors were activated and expanded in G-Rex bioreactors for 9 days and assessed for fold expansion. Data is the average of 4 independent experiments with 3 donors per experiment, error bars ±SD.

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

ProPak™ Recombinant Human IL-7 GMP Protein, CF Summary

• Details of Functionality: Measured in a cell proliferation assay using PHA-activated human peripheral blood lymphocytes (PBL). Yokota, T. et al. (1986) Proc. Natl. Acad. Sci. USA 83:5894. The ED₅₀ for this effect is 0.100-0.500 ng/mL. The specific activity of Recombinant Human IL-7 is >1.00 x 10⁸ units/mg, which is calibrated against the human IL-7 reference standard (NIBSC code: 90/530).
• Source: E. coli-derived human IL-7 protein
  Asp26-His177, with an N-terminal Met
  Produced using non-animal reagents in an animal-free laboratory.
  Manufactured and tested under cGMP guidelines.
• Accession #: P13232.1
• N-terminal Sequence: Met-Asp26-(Cys)-Asp-Ile-Glu-Gly-Lys-Asp-Gly
• 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 17507 Da ± 50 Da.
• Endotoxin Note: <5.0 EU/mL by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 17 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: 17 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  
  A minimum of 6 months when stored between -14 °C and -40 °C. Can be stored up to 2 weeks at 2-8 °C. Refer to lot specific COA for the Use by Date.
• Buffer: Supplied as a 0.2 μm filtered solution in PBS, recombinant HSA, and Trehalose.
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 17507 Da ± 50 Da.

Notes

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For preclinical, or clinical ex vivo use
Not for in vivo use

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ProPak™ Recombinant Human IL-7 GMP Protein, CF

• IL7
• IL-7
• IL-7interleukin-7
• interleukin 7
• Lymphopoietin-1
• PBGF

Background

IL-7 (interleukin-7) is a 25 kDa cytokine of the hemopoietin family that plays important roles in lymphocyte differentiation, proliferation, and survival (1-4). Human IL‑7 cDNA encodes 177 amino acids (aa) that include a 25 aa signal peptide (3). Human IL-7 shares approximately 60-63% aa sequence identity with mouse, rat, canine and feline IL-7, and 72-76% with equine, bovine, ovine, and porcine IL-7. Human and mouse IL-7 exhibit cross-species activity (2, 3). 

IL-7 is produced by a wide variety of cells in primary and secondary lymphoid tissues, including stromal epithelial cells of the thymus, bone marrow, and intestines (1, 2, 5). Circulating IL-7 is limiting in healthy animals, but increases during lymphopenia (1, 6). IL-7 signals through a complex of the IL-7 Receptor alpha subunit (IL-7 R alpha , also known as CD127) with the common gamma chain ( gamma c) (1). The gamma c is also a subunit of the receptors for IL-2, -4, -9, -15, and -21 (1). 

IL-7 R alpha is expressed on double negative (CD4-CD8-) and single positive (CD4+ or CD8+) naïve and memory T cells, but undergoes IL-7-mediated down‑regulation and shedding during antigen-driven T cell proliferation, and is absent on regulatory T cells (1, 2, 6-11). IL-7 contributes to the maintenance of all naïve and memory T cells, mainly by promoting expression of the anti-apoptotic protein Bcl-2 (9-11). It is required for optimal T cell-dendritic cell interaction (6). IL-7 is expressed early in B cell development prior to the appearance of surface IgM (1, 5, 9). In mouse, IL-7 activation of IL-7 R alpha is critical for both T cell and B cell lineage development, while in humans, it is required for T cell but not for B cell development (4, 9, 12, 13). However, IL-7 functions in both mouse and human pro-B cells to suppress premature Ig light chain recombination during proliferative growth (14, 15).

Like other common gamma-chain cytokines like IL-2 and IL-15, IL-7 and its receptor, IL-7R, has been used in a variety of immunotherapy applications, often in fluid tumors and in some instances of solid tumor models (16). Sometimes use of recombinant IL-7 is preferential as current studies and early clinical trials of cancer have found less severe toxicity or side effects upon treatment with IL-7 in comparison to IL-15 or IL-2 (16). 

In CAR-T cell therapies, enhanced expression and secretion of human IL-7 and CCL19 have enhanced the ability of T cells to expand and migrate in vitro (17). Engineered CAR T cells expressing IL-7 or a constitutively active IL-7R results in increased efficacy of CAR T anti-tumor effects (16, 18). IL-7 is also frequently used in combination with IL-15 as a supplement in cell culture of CAR T cells to support their expansion (19). Additionally, IL-7/IL-15 in the presence of cord blood-derived T cells helps to maintain their early differentiation state (20). Monoclonal antibodies against IL-7R or small molecule inhibitors against the IL-7R signaling pathway are commonly used in circumstances of autoimmune diseases to delay disease progression (16).  Also due to its ability to stimulate both adaptive and innate immune cells, treatment with IL-7 has shown improved survival in patients with sepsis who are at risk of deadly secondary infections (21), providing evidence for IL-7 applications beyond cancer immunotherapy.

1. Sasson, S.C. et al. (2006) Curr. Drug Targets 7:1571.
2. Barata, J.T. et al. (2006) Exp. Hematol. 34:1133.
3. Goodwin, R.G. et al. (1990) Proc. Natl. Acad. Sci. USA 86:302.
4. Namen, A.E. et al. (1988) Nature 333:571.
5. Shalapour, S. et al. (2012) PLoS ONE 7: e31939.
6. Saini, M. et al. (2009) Blood 113:5793.
7. Park, J.H. et al. (2004) Immunity 21:289.
8. Vranjkovic, A. et al. (2007) Int. Immunol. 19:1329.
9. Sudo, T. et al. (1993) Proc. Natl. Acad. Sci. 90:9125.
10. Seddon, B. et al. (2003) Nat. Immunol. 4:680.
11. Schluns, K.S. et al. (2000) Nat. Immunol. 5:426.
12. Peschon, J.J. et al. (1994) J. Exp. Med. 180:1955.
13. Pribyl, J.A. and T.W. LeBien (1996) Proc. Natl. Acad. Sci. 93:10348.
14. Johnson, K. et al. (2012) J. Immunol. 188:6084.
15. Nodland, S.E. et al. (2011) Blood 118:2116.
16. Wang, C. et al. (2022) Int. J. Mol. Sci. 23:10370.
17. Pang, N. et al. (2021) J Hematol Oncol. 14:118.
18. Li, L. et al. (2022) Sci Rep. 12:12506.
19. Xu, Y. et al. (2014) Blood. 123:3750.
20. Marton,C. et al. (2022) Cancer Gene Ther. 29:961.
21. Winer, H. et al. (2022) Cytokine. 160:156049.

Bioinformatics

• Uniprot:
  • P13232.1()