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Enhanced TNF Ligands

The critical role of signals transmitted via TNF ligands and receptors in mediating inflammation, cancer, and immunity has been well established in the literature. This understanding has been harnessed on the drug development frontier to mediate the pro-inflammatory properties of TNF signaling using ligand-neutralizing antibodies (eg. Remicade®) or soluble receptors (eg. Enbrel®). Similar strategies have sought to use recombinant TNF ligands to enhance, for instance, anti-viral and anti-cancer immune activation.

One limitation of this strategy is the inherent instability of recombinant ligands, and their inability to effectively mimic the natural clustering of TNF receptors on the cell surface required for immune activation. Oligomerization of TNF ligands using enhancers (eg. cross-linking antibodies), and expression of self-oligomerizing chimeric proteins (eg. Enzo MEGA® ligands) have been shown to improve stability and significantly enhance immune activation compared to that achieved with recombinant ligands alone (see figure below).

 

High-quality Enhanced Ligands Deliver Consistent Results

Enzo Life Sciences offers a diverse collection of enhanced ligands for use in immune activation studies. Our experience in the design and large-scale production of complex, high-purity, low endotoxin protein constructs results in enhanced ligands with the purity, stability, and consistency to deliver results you can trust for the duration of your study.

Oligomerization Domains in Enzo Enhanced Ligands:
  • MEGA® Ligands – use the ACRP30headless domain from human or mouse ACRP30
  • KILLER™ Ligands – use a proprietary linker peptide that promotes trimerization
  • SUPERKILLER™ Ligands – use the KILLER™ linker peptide mutated to increase disulfide-mediated cross-linking

Enhance Immune Activation with MEGACD40L® Protein

B cell lymphocyte activation by various CD40L constructs. PBMCs were treated for 48 hrs in serum free media containing serially diluted MEGACD40L® Protein (ALX-522-110), CD40L + 2ug/mL Enhancer, or CD40L. Cells were dual-stained with anti-human CD19–PE and anti-human CD86–APC and analyzed by flow cytometry. Data are presented as the percent of CD19+ B cells that co-stain as CD86+.

Featured Product

Enhanced ligand that utilizes the KILLER™ linker peptide mutated to increase disulfide-mediated cross-linking to form a more stable oligomer
  • Improves stability
  • Enhances immune activation compared to other recombinant ligands


Figure: Comparison of pro-apoptotic activities of human and mouse SUPERKILLERTRAIL™ Protein, Soluble (human) (rec.) on human tumor cell lines Jurkat and CEM.
Method: Cells were treated with mouse and human SUPERKILLERTRAIL™ Protein, Soluble (human) (rec.) (CC-mutant) (50ng/ml) for 14 hours, stained with propidium iodide and analyzed by flow cytometry.
 

Featured Enhanced TNF Ligands:

MEGACD40L® Protein (soluble) (human), (recombinant) 

High activity, high purity CD40L protein for co-stimulatory activation of an immune response
Produced in CHO cells. The extracellular domain of human CD40L (CD154) (aa 116-261) is fused at the N-terminus to mouse ACRP30headless (aa 18-111) and a FLAG®-tag., ≥90% (SDS-PAGE) | Print as PDF
 
ALX-522-110-C010 10 µg 499.00 USD
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MEGACD40L® Protein (soluble) (mouse), (recombinant) 

Highly active
Produced in CHO cells. The extracellular domain of mouse CD40L (CD154) (aa 115-260) is fused at the N-terminus to mouse ACRP30headless (aa 18-111) and a FLAG®-tag., ≥95% (SDS-PAGE) | Print as PDF
 
ALX-522-120-C010 10 µg 536.00 USD
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SUPERFASLIGAND® Protein (soluble) (human), (recombinant) 

Fas ligand with improved stability providing significantly enhanced immune activation.
Produced in HEK 293 cells. The extracellular domain of human FasL (APO-1L; CD95L; CD178) (aa 103-281) is fused at the N-terminus to a linker peptide (26 aa) and a FLAG®-tag. Glycosylation of rhs SUPERFASLIGAND® is similar to natural human FasL., ≥95% (SDS-PAGE), ELISA | Print as PDF
 
ALX-522-020-C005 5 µg 284.00 USD
 
ALX-522-020-3005 3x5 µg SuperPack 657.00 USD
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KillerTRAIL™ Protein (soluble) (human), (recombinant) 

Enhanced ligand that utilizes the proprietary Killer™ linker peptide that promotes stability.
Produced in E. coli. The extracellular domain of human TRAIL (aa 95-281) is fused at the N-terminus to a His-tag and a linker peptide., ≥99% (SDS-PAGE, MS analysis) | Print as PDF
 
ALX-201-073-C020 20 µg 444.00 USD
 
ALX-201-073-3020 3x20 µg 913.00 USD
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SUPERKILLERTRAIL® Protein (soluble) (human), (recombinant) 

Enhanced ligand with improved stability providing significantly enhanced immune activation.
Produced in E. coli. The extracellular domain of human TRAIL (aa 95-281) is fused at the N-terminus to a His-tag and a linker peptide. The active multimeric conformation is stabilized by an inserted mutation allowing an additional CC-bridge., ≥98% (SDS-PAGE). MS analysis, <1% impurity (mainly Hsp70 protein from E. coli) | Print as PDF
 
ALX-201-115-C010 10 µg 444.00 USD
 
ALX-201-115-3010 3x10 µg 940.00 USD
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ACRP30headless (human), (recombinant) 

Produced in HEK 293 cells. The non-homologous (aa 16-45) and collagen domain of human ACRP30 (aa 45-108) are fused at the N-terminus to a FLAG®-tag., ≥90% (SDS-PAGE) | Print as PDF
 
ALX-203-002-C010 10 µg 281.00 USD
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ACRP30headless (mouse), (recombinant) 

Produced in HEK 293 cells. The collagen domain of mouse ACRP30 (aa 18-111) is fused at the N-terminus to a linker peptide (8 aa) and a FLAG®-tag., ≥90% (SDS-PAGE) | Print as PDF
 
ALX-203-003-C010 10 µg 281.00 USD
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TNF-α (human), (recombinant) (cell culture grade) 

94948-59-1, Produced in yeast., ≥97% (SDS-PAGE) | Print as PDF
 
ALX-520-002-C010 10 µg 184.00 USD
 
ALX-520-002-C050 50 µg 655.00 USD
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Fc (human):TNF-α (soluble) (human), (recombinant) 

Produced in CHO cells. The extracellular domain of human TNF-α (aa 85-233) is fused at the N-terminus to the Fc portion of human IgG1 and a linker peptide (20 aa)., ≥90% (SDS-PAGE) | Print as PDF
 
ALX-522-087-C010 10 µg 426.00 USD
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