MACS GMP Recombinant Human Interleukin-12 (IL-12) is a recombinant protein optimized as an ancillary material for
ex vivo
cell processing. It is not intended for human
in vivo
applications.

Data and images for
MACS
®
GMP Recombinant Human IL-12

Figures

Figure 1

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MACS GMP Recombinant Human IL-12 biological activity.
Activity of Human MACS GMP IL-12 (blue bar), was compared to another commercially available product (black bar).

Figure 1

MACS GMP Recombinant Human IL-12 biological activity.
Activity of Human MACS GMP IL-12 (blue bar), was compared to another commercially available product (black bar).

Figure

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The biological activity of MACS GMP Recombinant Human IL-12 was determined by induction of IFN-γ secretion by PHA-activated T cells.
The biological activity of MACS GMP Recombinant Human IL-12 was determined by induction of IFN-γ secretion by PHA-activated T cells.

Specifications for
MACS
®
GMP Recombinant Human IL-12

Overview

MACS GMP Recombinant Human Interleukin-12 (IL-12) is a recombinant protein optimized as an ancillary material for
ex vivo
cell processing. It is not intended for human
in vivo
applications.

Detailed product information

Background information

IL-12 is a heterodimeric cytokine composed of two subunits (p35 and p40), which are encoded by two different genes, IL-12A (p35) and IL-12B (p40). The complete heterodimeric protein complex is termed IL-12p70. IL-12 is the most prominent member of the IL-12 cytokine family, which also contains other heterodimeric cytokines such as IL-23, IL-27 and IL-35
6
.
The p35 subunit of IL-12 contains 4 α-helix bundles, while the p40 subunit is structurally related to soluble IL-6 receptor alpha (sIL-6Rα). Hence, the structure of IL-12 is in essence that of a cytokine bound to a soluble receptor, and the ability to secrete the cytokine-like subunit (p35) depends upon association with the receptor-like subunit (p40)
7
.
IL-12 can affect many cell types, but its influence on T-cell-dependent responses has been most extensively studied: It is secreted by professional antigen presenting cells (mostly dendritic cells, but also macrophages and B cells) upon activation by microbial pathogens and binds to the IL-12 receptor complex (comprised IL-12Rβ1 and IL-12Rβ2) on T and NK cells. Receptor engagement induces the upregulation of transcription factor T-bet in CD4
+
and CD8
+
T cells and promotes their differentiation into Th1 cells or CTLs respectively. These activated T cells then secrete IFNγ and TNFα. IL-12 can also induce IFNγ production by NK cells
7
.
Naïve T cells express the IL-12 receptor subunit IL-12Rβ1, however expression of IL-12Rβ2 - and thereby responsiveness of naïve T cells to IL-12 - is dependent on TCR engagement, co-stimulation by CD28 and IL-2 signaling
8
. Signaling through the IL-12 receptor seems to be necessary for full-fledged T cell effector functions of the T cells and for IL-2 dependent proliferation
2
.

Applications

MACS GMP Recombinant Human IL-12 can be used for a variety of applications, including:
Optimization of
ex vivo
expansion protocols for adoptive cell transfer. Studies have demonstrated that
ex vivo
activation of T cells in the presence of IL-12 can improve expansion and effector function and prevent
in vivo
T cell exhaustion in animal models.
1
Generation of cytokine-induced memory-like natural killer cells (CIML NK). Memory-like NK cells can be generated
ex vivo
by cultivating human NK cells with a combination of IL-12, IL-15 and IL-18. Such CIML NK cells demonstrated an increased longevity and enhanced anticancer functionality compared to normal effector NK cells.
2
In vitro
differentiation of naive CD4
+
T cells towards Th1 cells. After activation, naïve CD4
+
T cells can be differentiated into Th1 effector cells when cultured with a combination of IL-2, IL-12 and IFNγ.
3
Technical specifications
MACS GMP Recombinant Human IL-12 is manufactured and released based on the stringent specifications displayed below.
Lot-specific results of the respective QC measurements are depicted on each Product Quality Certificate (PQC). Learn more
here.
  • Identity: Confirmed by isoelectric focusing and chip electrophoresis. This corresponds to the mature form of IL-12.
  • Purity: ≥ 95 % as determined by chip electrophoresis.
  • Product-related proteins: ≤15% as determined by chip electrophoresis.
  • Endotoxin content: < 50 EU/mg, as determined by kinetic Limulus Amoebocyte Lysate (LAL) assay (Pharmacopoeia Europaea (Ph. Eur.)).
  • Residual host cell DNA content: < 20 ng/mg, as determined by quantitative PCR specific for CHO genomic DNA.
  • Residual host cell protein content: < 25 µg/mg, as determined by CHO HCP ELISA.
MACS GMP Recombinant Human IL-12 is expressed as a single chain fusion protein. It has been shown that such fusion proteins of naturally occurring heterodimeric cytokines such as IL-12 or IL-23 are bioactive
in vitro
and
in vivo
.
5
The product is lyophilized without carrier protein or preservatives.

Biological activity

  • The specific activity is determined by induction of IFN-γ secretion by PHA-activated T cells.
    1
    The proliferation assay was calibrated with the reference reagent for human IL-12 (NIBSC code 95/544) provided by the WHO/National Institute for Biological Standards and Control.
  • Minimal specific activity: ≥ 3×
    10
    6
    U/mg

Quality statement

MACS GMP Products are manufactured and tested under a quality management system (ISO 13485) and are in compliance with relevant GMP guidelines. They are designed following the recommendations of USP <1043> on ancillary materials.

Disclaimer

MACS GMP Products are for
ex vivo
cell processing only, and are not intended for human
in vivo
applications. For regulatory status in the USA, please contact your local representative.

Resources for
MACS
®
GMP Recombinant Human IL-12

Certificates

Please follow this
link
to search for Product Quality Certificates (PQC) by lot number.

References for
MACS
®
GMP Recombinant Human IL-12

Publications

  1. Tucker, C. G. et al. (2020) Adoptive T Cell Therapy with IL-12-Preconditioned Low-Avidity T Cells Prevents Exhaustion and Results in Enhanced T Cell Activation, Enhanced Tumor Clearance, and Decreased Risk for Autoimmunity. J Immunol 205(5): 1449-1460
  2. Tarannum, M. et al. (2021) Cytokine‑induced memory‑like natural killer cells for cancer immunotherapy. Stem Cell Res Ther. 12: 592
  3. Wulff, H. et al. (2007) Cloning and characterization of an adenoviral vector for highly efficient and doxycycline-suppressible expression of bioactive human single-chain interleukin 12 in colon cancer. BMC Biotechnol. 7: 35
  4. Lieschke, G. J. et al. (1997)
    Bioactive murine and human interleukin-12 fusion proteins which retain antitumor activity
    in vivo.
    Nat. Biotechnol. 15: 35-40
  5. Vignali, D. et al. (2014) IL-12 Family Cytokines: Immunological Playmakers. Nat Immunol 13(8): 722-728
  6. Wojno, E. D. et al. (2019) The Immunobiology of the Interleukin-12 Family: Room for Discovery. Immunity 50(4): 851-870
  7. Valenzuela, J. et al. (2002) The Roles of IL-12 in Providing a Third Signal for Clonal Expansion of Naive CD8 T Cells. J Immunol 169(12): 6842-6849

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®
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