Recombinant human IL-23 protein - Bioactive cytokine

Human IL-23 (linked heterodimer) protein - Mammalian cell-expressed, tag-free, carrier-free

Recombinant human IL-23 is a high-quality and biologically active cytokine, validated using proprietary IL-23 reporter cells. This pro-inflammatory cytokine comprises two subunits: a specific IL-23p19 subunit and a common IL-12p40 subunit shared with IL-12. It is produced in CHO cells to ensure protein glycosylation and bona fide 3D structure.

Recombinant human IL-23 can be used together with HEK-Blue™ IL-23 cells for the screening of inhibitory molecules, such as Ustekinumab, a therapeutic monoclonal antibody targeting the IL-12/IL-23 p40 subunit (see figures).

IL-23 signaling and biological functions

Key features

• Each lot is validated using HEK-Blue™ IL-23 cells 
• Endotoxin < 0.1 EU/µg
• 0.2 µm sterile-filtered

Applications

• IL-23 detection and quantification assays (positive control)
• Screening and release assays for antibodies blocking IL-23 signaling
• Screening and release assays for engineered IL-23 proteins

Interleukin 23 (IL-23) is a pro-inflammatory cytokine driving IL-17, IFN-γ, and IL-22 production by Th17 and group 3 innate lymphoid (ILC3) cells in response to microbial infections at barrier sites. Dysregulated IL-23 production is linked to immune-mediated inflammatory diseases such as psoriasis, Crohn's disease, and ulcerative colitis. 

 More details

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

Figures

SDS-PAGE analysis of the recombinant human (h)IL-23. 1 µg of hIL-23 was loaded on a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad). Detection was performed as per the manufacturer’s instructions. A band was detected at ~61 kDa.

Dose-response of HEK-Blue™ IL-23 cells to recombinant human (h)IL-23. Cells were stimulated with increasing concentrations of recombinant human IL-23. After overnight incubation, the STAT3-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™ IL-12 or HEK-Blue™ IL-23 cells response using Ustekinumab biosimilar. Increasing concentrations of Anti-hIL-12/IL-23-p40-hIgG1 (0.3 ng/ml - 30 µg/ml) were incubated with recombinant human (h)IL-12 (3 ng/ml) or hIL-23 (1 ng/ml) for 1 h before the addition of either HEK-Blue™ IL-12 cells (A) or HEK-Blue™ IL-23 cells (B). 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

Carrier: Carrier-free

Tag: Tag-free

Construction: Heterodimer linked by 10 amino acids (a.a.)

Accession number: P29460.1 (IL-12p40), Q9NPF7 (IL-23p19)

Protein size: 486 a.a. (I23-S328(p40)-Linker-R20-P189(p19))

Molecular weight: ~ 61 kDa (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 IL-23 is provided lyophilized and is available in two quantities:

• rcyc-hil23: 10 µg
• rcyc-hil23-5: 5 x 10 µg
• 1.5 ml endotoxin-free water for rcyc-hil23 and rcyc-hil23-5

 Recombinant IL-23 is shipped at room temperature.

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

 Avoid repeated freeze-thaw cycles.

Details

IL-23 background

Interleukin 23 (IL-23) belongs to the heterodimeric IL-12 cytokine family. This family is characterized by the sharing of p19, p28, p35, p40, and Ebi3 subunits, and includes IL-12, IL-23, IL-27, IL-35, and IL-39 [1]. The secretion of bioactive IL-23 requires the co-expression of IL-23p19 and IL-12p40 and disulfide bond formation. IL-23 is primarily produced by pathogen-activated macrophages and dendritic cells (DC) in barrier tissues, such as the gut and skin [1].

The binding of IL-23 to its IL-12Rβ1/IL-23R heterodimeric receptor triggers a JAK2/TYK2 signal transduction, leading to the activation of STAT3 or STAT4. Activated STAT3 and STAT4 form homodimers that are translocated to the nucleus, where they regulate the expression of target genes. The IL-23 receptor is expressed by activated Th17 cells and activated group 3 innate lymphoid cells (ILC3) [2]. IL-23 signaling promotes the secretion of IL-17A and IFN-γ by pathogenic Th17 cells and IL-17A, IL-17F, and IL-22 by ILC3 cells [2]. Moreover, IL-23 signaling in these cells promotes the expression of IL-23R in a positive feedback loop.

Relevance for therapeutics development

IL-23 is a key cytokine for controlling microbial infections at barrier sites [3]. However, excessive IL-23 production contributes to immune-mediated inflammation diseases, including psoriasis and IBD (Inflammatory bowel disease, encompassing Crohn's disease and ulcerative colitis) [2, 4]. Additionally, genetic studies showed an association between IL-23R polymorphism and IBD [4].

Ustekinumab is a fully human monoclonal antibody (mAb) that targets the p40 subunit common to IL-12 and IL-23, preventing their interactions with their receptor common chain IL-12Rβ1 [5]. This antibody is FDA-approved for treating psoriasis, psoriatic arthritis, Crohn's disease, and ulcerative colitis [6].

Risankinumab and Mirikizumab are two human mAbs targeting the IL-23p19 specific subunit of IL-23, blocking the cytokine interaction with its receptor [4, 7]. Risankinumab is FDA-approved for treating plaque psoriasis, psoriatic arthritis, and Crohn's disease in adults [8]. Mirikizumab is FDA-approved as a second-line therapy for moderate-to-severe active ulcerative colitis [9].

Addressing the benefits of neutralizing IL-12 and IL-23 together or individually, depending on the clinical context, can be achieved using neutralizing mAbs targeting the p19 subunit of IL-23 [3].

References:

1. Floss, D.M., et al., 2020. IL-12 and IL-23-Close Relatives with Structural Homologies but Distinct Immunological Functions. Cells, 9(10): 2184.
2. Moschen, A.R., et al., 2019. IL-12, IL-23 and IL-17 in IBD: immunobiology and therapeutic targeting. Nat rev Gastroenterol Hepatol. 16(3):185-196.
3. Tait Wojno, E.D. et al., 2019. The Immunobiology of the Interleukin-12 Family: Room for Discovery. Immunity. 50(4):851-870.
4. Parigi T.L., et al., 2022. Blockade of IL-23: What is in the pipeline? J Crohns Colitis. 16 (supp 2):ii64-ii72.
5. Benson, J.M, et al., 2011. Discovery and mechanism of ustekinumab: a human monoclonal antibody targeting interleukin-12 and interleukin-23 for treatment of immune-mediated disorders. MAbs 3(6):535-545.
6. FDA website accessed in Nov 2024: https://www.accessdata.fda.gov/drugsatfda_docs/label/2024/761044s013lbl.pdf#page=37
7. Danese, S. & Peyrin-Biroulet, L., 2024. Are all IL-23 blockers the same in inflammatory disease? Nat Rev Gastr & Hepatol. 21(3):138-139.
8. https://www.accessdata.fda.gov/drugsatfda_docs/label/2024/761105s029,761262s007lbl.pdf
9. https://pi.lilly.com/us/omvoh-uspi.pdf?s=pi