Recombinant human IL-17A protein - Bioactive cytokine

Human IL-17A protein - Mammalian cell-expressed, tag-free, carrier-free

Recombinant human IL-17A is a high-quality and biologically active cytokine, validated using proprietary IL-17 reporter cells. This pro-inflammatory cytokine is produced in CHO cells to ensure protein glycosylation and bona fide 3D structure.

Recombinant human IL-17A can be used together with HEK-Blue™ IL-17 cells for the screening of inhibitory molecules, such as Secukinumab, a therapeutic monoclonal antibody targeting IL-17A (see figures).

IL-17A signaling and biological functions

Key features

• Each lot is validated using HEK-Blue™ IL-17 cells
• Endotoxin ≤ 0.001 EU/µg
• 0.2 µm sterile-filtered

Applications

• Standard for IL-17A detection and quantification
• Screening and release assays for antibodies blocking IL-17A signaling
• Screening and release assays for engineered IL-17

The interleukin-17 (IL-17) family comprises six members (IL-17A – 17F), which have various biological functions, including driving an inflammatory cascade during infections and autoimmune diseases. IL-17A is a key therapeutic target for the treatment of moderate-to-severe plaque psoriasis, psoriatic arthritis, and ankylosing spondylitis.

 More details

All InvivoGen products are for internal research use only and not for human or veterinary use.

Figures

SDS PAGE of the recombinant human (h)IL-17A protein. 1 μg of CHO-derived hIL-17A was loaded on a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad). Detection was performed as per the manufacturer’s instructions. Its predicted size is ~15 kDa (lower band). The higher band at ~18 kDa corresponds to the glycosylated protein. 

Dose-response of HEK-Blue™ IL-17 cells to recombinant human IL-17A. HEK-Blue™ IL-17 cells were stimulated with increasing concentrations of recombinant IL-17A. After overnight incubation, the NF-kB/AP-1-induced SEAP activity was determined using QUANTI-Blue™ Solution, a SEAP detection reagent. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

Dose-dependent inhibition of HEK-Blue™ IL-17 cell response using Secukinumab biosimilar. Increasing concentrations of Anti-hIL-17-hIgG1 (0.1 ng/ml - 10 µg/ml) were incubated with recombinant human IL-17A (1 ng/ml) for 1 hour prior to the addition of HEK-Blue IL-17 cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown as the percentage of activity (mean ± SEM).

Specifications

Source: Chinese hamster ovary (CHO) cells

Species: Human

Carrier: Carrier-free

Tag: Tag-free

Accession number: Q16552

Protein size (predicted): 132 a.a.  (G24-A155)

Molecular weight: ~ 16 kDa (SDS-PAGE)

Solubility: 100 μg/ml in water

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

Form:  Lyophilized

Reconstitution buffer:  Endotoxin-free water (provided)

Purity:  ≥95% (SDS-PAGE)

Endotoxin: ≤ 0.001 EU/μg (measurement by kinetic chromogenic LAL assay)

Tested applications:  Cellular assays

Quality control: Each lot is functionally tested and validated.

Contents

Recombinant human IL-17A is provided lyophilized and is available in two quantities:

• rcyc-hil17a: 20 µg
• rcyc-hil17a-5: 5 x 20 µg (100 µg)
• 1.5 ml endotoxin-free water for rcyc-hil17a and rcyc-hil17a-5

 Recombinant hIL-17A is shipped at room temperature.

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

 Avoid repeated freeze-thaw cycles.

Details

IL-17A background

Interleukin 17 (IL-17) is a family of six closely related cytokines (IL-17A to IL-17F) which have both pro-inflammatory and anti-inflammatory activities. IL-17A and IL-17F, which can form a heterodimer, play an important role in Th17 immunity and are implicated in tumorigenesis and autoimmune diseases, whereas IL-17E (also known as IL-25) appears to promote Th2 immunity [1,2]. Interleukin‑17A (IL‑17A), also known as CTLA‑8, is a 15‑20 kDa glycosylated cytokine that plays an important role in anti‑microbial and chronic inflammation. It induces chemokine production, neutrophil influx, and the production of antibacterial peptides [3].

IL-17 cytokines exert their biological activities by binding to heterodimeric receptors containing the ubiquitous IL-17RA chain and a second IL-17R(C, B, or E) chain. IL-17A and IL-17F bind to the IL-17RA/IL-17RC receptor, IL-17C binds to the IL-17RA/IL-17RE receptor, and IL-17E binds to the IL-17RA/IL-17RB receptor [1, 2]. The activated heterodimeric receptor recruits the Act1 adaptor and induces the TNF receptor-associated factor 6 (TRAF6) ubiquitylation. This triggers a signaling cascade that results in NF-κB and AP-1 activation [1].

Antibodies targeting IL-17A, namely Secukinumab and Ixekizumab, were approved in 2016 for the treatment of moderate-to-severe plaque psoriasis, ankylosing spondylitis (AS), and psoriatic arthritis (PsA). Surprisingly, trials of Secukinumab and Brodalumab, an anti-IL-17RA in Crohn’s disease, were terminated early due to worsening of the disease in the treatment group [3-4]. These findings agree with IL-17's supportive role in skin wound healing [3].

References:

1. Monin L. & Gaffen S.L., 2018. Interleukin 17 family cytokines: signaling mechanisms, biological activities, and therapeutic implications. Cold Spring Harb Perspect Biol. 10(4).
2. Pappu R. et al., 2011. The interleukin-17 cytokine family: critical players in host defence and inflammatory diseases. Immunology. 134: 8-16.
3. Huangfu L, Li R, Huang Y, Wang S, 2023. The IL-17 family in diseases: from bench to bedside. Signal Transduct Target Ther.11;8(1):402.
4. Amatya N, Garg AV, Gaffen SL, 2017. IL-17 Signaling: The Yin and the Yang. Trends Immunol. 38(5):310-322.