Recombinant human TL1A/TNFSF15 protein - Bioactive cytokine

Human TL1A protein - Mammalian cell-expressed, tag-free, carrier-free

Recombinant human TL1A is a high-quality and biologically active cytokine, validated using proprietary TL1A reporter cells. This member of the TNF superfamily is produced in CHO cells to ensure protein glycosylation and bona fide 3D structure. Human TL1A produced in E. coli can form aggregates, potentially impairing its intracellular signaling efficiency (in-house observations).

Recombinant human TL1A can be used together with HEK-Blue™ TL1A cells for the screening of inhibitory molecules, such as Tulisokibart, a monoclonal antibody targeting TL1A (see figures).

TL1A signaling and biological functions

Key features

• Each lot is validated using HEK-Blue™ TL1A cells
• Endotoxin < 0.1 EU/µg
• 0.2 µm sterile-filtered

Applications

• Standard for TL1A detection and quantification
• Screening and release assays for antibodies blocking TL1A signaling
• Screening and release assays for engineered TL1A

Tumor necrosis factor (TNF)-like 1A (TL1A) drives the production of pro-inflammatory cytokines and differentiation of T helper subsets. Abnormal TL1A expression is linked to enteric autoimmune diseases (e.g. Crohn’s disease) and extends to other diseases including allergic airway inflammation.

 More details

InvivoGen’s products are for research use only, and not for clinical or veterinary use.

Figures

SDS-PAGE of the recombinant human (h)TL1A cytokine. 1.5 μg of hTL1A, treated or not with dithiothreitol (DTT) reducing agent, was loaded on a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad). Detection was performed as per the manufacturer’s instructions. Without DTT, multimeric bands were detected at ~50 kDa. In the presence of DTT, monomeric bands were detected at ~28 kDa. Note: As hTL1A is produced in mammalian cells, it displays heterogeneous glycosylation, thus impacting its size (arrows).

Dose-response of HEK-Blue™ TL1A cells to recombinant TL1A. Cells were stimulated with increasing concentrations of recombinant human (h) or mouse (m) TL1A. After overnight incubation, the NF-κB/AP1-induced SEAP activity was determined using QUANTI-Blue™, a SEAP detection reagent. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

 

Dose-dependent inhibition of HEK-Blue™ TL1A cell response using Tulisokibart biosimilar. Increasing concentrations of Anti-hTL1A-hIgG1 (0.01 ng/ml - 1 µg/ml) were incubated with recombinant human TL1A (3 ng/ml) for 1 h before the addition of HEK-Blue™ TL1A cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown in percentage of activity (mean ± SEM).

Specifications

Source: Chinese hamster ovary (CHO) cells

Species: Human

Alternative name: TNFSF15

Carrier: Carrier-free

Tag: Tag-free

Accession number: O95150

Protein size: 180 a.a. (L72-L251)

Molecular weight: ~ 24-27 kDa (monomer) (SDS-PAGE)

Solubility: 100 μg/ml in water

Formulation: Phosphate buffer saline (pH 7.4), 5% saccharose

Sterility: 0.2 µm filtration

Form:  Lyophilized

Reconstitution buffer:  Endotoxin-free water (provided)

Purity:  ≥95% (SDS-PAGE)

Endotoxin level:  ≤ 0.1 EU/μg (measurement by kinetic chromogenic LAL assay)

Tested applications:  Cellular assays

Quality control: Each lot is functionally tested and validated.

Contents

Recombinant human TL1A is provided lyophilized and is available in two quantities:

• rcyc-htl1a: 20 µg
• rcyc-htl1a-5: 5 x 20 µg
• 1.5 ml endotoxin-free water for rcyc-htl1a and rcyc-htl1a-5

 Recombinant hTL1A is shipped at room temperature.

 Upon receipt, the product should be stored at -20°C.

 Avoid repeated freeze-thaw cycles.

Details

TL1A background

Tumor necrosis factor (TNF)-like 1A (TL1A), also called TNFSF15, is a cytokine that belongs to the TNF superfamily. TL1A is produced by endothelial cells, activated dendritic cells, and macrophages. It is synthesized as a membrane-bound trimeric molecule. Alternative splicing or cleavage by the tumor necrosis factor-alpha converting enzyme results in soluble TL1A [1]. Both forms bind the homotrimeric transmembrane death receptor 3 (DR3), triggering TRADD/TRAF2/RIP signaling, and ultimately leading to NF-κB and MAPKs activation. Subsequent gene expression in target cells drives the production of pro-inflammatory cytokines and differentiation of T helper subsets [1].

Relevance for therapeutics development

TL1A is normally responsible for mediating protective immune responses. However, it can also play a role in chronic inflammatory disorders and have harmful impacts. Abnormal TL1A expression or TL1A overactivity promotes excessive inflammation and immune cell infiltration, contributing to tissue damage and further inflammation amplification [1].

Pharmaceutical companies have developed potent monoclonal antibodies to block TL1A signaling and stop the downstream inflammatory cascade and tissue damage, primarily in the onset of asthma and Crohn’s disease. Anti-TL1A inhibitors such as Tulisokibart (MK-7240 or PRA023, Merck), RVT-3101 (PF-06480605, Roche), and Duvakitug (TEV-48574, Sanofi & Teva Pharmaceuticals) have demonstrated "best-in-class" potential for treating inflammation and scarring in IBD [2-4]. Duvakitug is currently being studied for its safety as an asthma medication [5-6].

References:

1. Xu W.D., et al., 2022. Role of TL1A in Inflammatory Autoimmune Diseases: A Comprehensive Review. Front. Immunol. 13: 891328.
2. Sands, B.E., et al., 2024. Phase 2 Trial of Anti-TL1A Monoclonal Antibody Tulisokibart for Ulcerative Colitis. N Engl J Med, 391(12): p. 1119-1129.
3. Danese, S., et al., 2021. Anti-TL1A Antibody PF-06480605 Safety and Efficacy for Ulcerative Colitis: A Phase 2a Single-Arm Study. Clinical Gastroenterology and Hepatology. 19(11): p. 2324-2332.e6.
4. Solitano, V., et al., 2024. TL1A inhibition for inflammatory bowel disease treatment: From inflammation to fibrosis. Med. 5(5): p. 386-400.
5. Balyan, R., et al.,, 2024. P633 First-in-Human Pharmacokinetic and Safety Study of an Anti-TL1A Antibody, TEV-48574, in Healthy Volunteers and Asthma Patients. Journal of Crohn's and Colitis. 18(Supplement_1): p. i1215-i1216.
6. Raphael, G., et al., 2024. P1061 TEV-48574, an anti-TL1A antibody in development for use in IBD, is safe and well tolerated following 16 weeks of subcutaneous treatment in adults with severe uncontrolled T2-low/non T2 asthma. Journal of Crohn's and Colitis. 18(Supplement_1): p. i1908-i1908.