Recombinant Human IL-29

Recombinant human IL-29 (also designated as interferon-lambda 1, IFN-λ1) offers several compelling advantages for research applications across immunology, virology, and oncology.

Cellular Specificity and Reduced Toxicity

A primary advantage of IL-29 is its selective targeting of epithelial tissues and hepatic cells while avoiding toxic effects on endothelial cells in the central nervous system, kidney, and spleen. This cell-type selectivity distinguishes IL-29 from Type I interferons, which affect multiple cell populations broadly. This specificity is particularly valuable when you need to study epithelial-specific immune responses without confounding systemic effects.

Potent Antiviral Activity

IL-29 demonstrates robust antiviral properties against diverse viral pathogens. The cytokine exhibits particularly strong activity in vivo compared to in vitro conditions, and it provides antiviral defense through mechanisms independent of Type I interferons. This makes IL-29 especially useful for studying hepatitis C virus (HCV) infection and other viral diseases where epithelial tissues are primary targets. Additionally, IL-29 enhances interferon-gamma serum levels, amplifying the antiviral immune response.

Dual Immunomodulatory Functions

IL-29 regulates both innate and adaptive immunity. The cytokine upregulates interferon-stimulated genes and toll-like receptor expression in B cells, enhancing TLR7/8-mediated antibody production. This dual functionality makes IL-29 valuable for investigating complex immune mechanisms and developing immunotherapeutic strategies.

Antitumor Potential

IL-29 exhibits anti-proliferative and antitumor activities against various cancer cell lines, including oesophageal carcinoma, gastric adenocarcinoma, and ileocecal adenocarcinoma. Importantly, unlike Type I interferons, IL-29 selectively targets cancer cells while sparing normal cells, reducing off-target toxicity in your experimental models. Engineered IL-29 variants created through site-directed mutagenesis display enhanced anti-tumor activities, offering opportunities for optimized therapeutic development.

Therapeutic Development Platform

IL-29 is currently being developed as a clinical biotherapeutic, making recombinant preparations valuable for translational research. The well-characterized nature of this cytokine and its established bioactivity in standardized assays facilitate reproducible experimental design and comparison across studies.

Yes, recombinant human IL-29 can be used as a standard for quantification or calibration in ELISA assays, provided it is validated for this purpose in your specific assay system. Most commercial human IL-29 ELISA kits use recombinant IL-29 as the standard for generating calibration curves, enabling quantification of both endogenous (natural) and recombinant IL-29 in samples.

Key points and best practices:

• Assay Compatibility: ELISA kits for human IL-29 are typically validated to detect and quantify both natural and recombinant forms of IL-29. The standard curve is usually generated using recombinant IL-29, and the assay is calibrated accordingly.
• Standard Preparation: Prepare the recombinant IL-29 standard according to the instructions provided with your ELISA kit or assay protocol. This usually involves serial dilution in the recommended assay buffer to cover the dynamic range of the assay.
• Validation: Ensure that the recombinant IL-29 you use as a standard is of high purity and bioactivity, and that it matches the sequence and post-translational modifications (if relevant) of the IL-29 detected by your assay antibodies. Most kits specify that their antibodies recognize both natural and recombinant IL-29, but if you are developing your own assay, confirm cross-reactivity and recovery.
• Matrix Effects: If you are quantifying IL-29 in complex biological matrices (e.g., serum, plasma, cell culture supernatant), validate recovery by spiking known amounts of recombinant IL-29 into your sample matrix and confirming accurate quantification.
• Documentation: Refer to your ELISA kit’s manual for specific instructions on standard preparation and validation. Many protocols explicitly state the use of recombinant IL-29 as the standard and provide recovery and linearity data to support its use.

Summary Table: Use of Recombinant Human IL-29 as ELISA Standard

In conclusion: Using recombinant human IL-29 as a standard is standard practice for quantification in ELISA assays, as long as the assay is validated for both recombinant and natural forms and the standard is prepared according to protocol.

Recombinant Human IL-29 (IFN-λ1) has been validated in published research for several key applications, primarily related to its biological activity and immunological functions. The main applications include:

• Bioassay: IL-29 is frequently used in bioassays to assess its antiviral, antitumor, and immunomodulatory activities. For example, it has been used to study antiviral responses in human whole cells, including epithelial cells and hepatocytes, and to evaluate its effects on host defense mechanisms and immune signaling pathways.

• Cell Culture: IL-29 is applied in cell culture systems to stimulate cells and investigate its effects on signaling pathways, such as NF-κB activation, interferon signaling, and cytokine production. It is used to study the regulation of immune responses and the impact on cell proliferation and survival.

• Stimulation: IL-29 is used as a stimulant to activate immune cells and epithelial cells, allowing researchers to study the expression of interferon-stimulated genes (ISGs) and the regulation of immune responses.

• Control: In some studies, IL-29 serves as a control cytokine to compare the effects of other interferons or cytokines on viral replication and immune responses.

• Functional Studies: IL-29 is used in functional studies to investigate its role in mucosal immunity, antiviral defense, and the regulation of immune responses at epithelial barriers.

These applications highlight the versatility of Recombinant Human IL-29 in research settings, particularly in immunology and virology.

To reconstitute and prepare Recombinant Human IL-29 protein for cell culture experiments, dissolve the lyophilized protein in sterile buffer—typically phosphate-buffered saline (PBS), pH 7.4, containing 0.1% endotoxin-free recombinant human serum albumin (HSA) or bovine serum albumin (BSA)—to a concentration of 100–200 μg/mL.

Step-by-step protocol:

• Centrifuge the vial briefly before opening to ensure all lyophilized material is at the bottom.
• Add sterile PBS (pH 7.4) containing 0.1% HSA or BSA to the vial to reach your desired concentration (commonly 100–200 μg/mL).
• Gently swirl or tap the vial to mix. Avoid vigorous pipetting or vortexing, which can denature the protein.
• Let the solution stand at room temperature for 10–15 minutes to ensure complete dissolution.
• Aliquot the reconstituted protein into single-use volumes to avoid repeated freeze-thaw cycles, which can reduce activity.
• Store aliquots at –20°C to –70°C for long-term storage. After reconstitution, the protein is typically stable for up to 1 month at 2–8°C or 3 months at –20°C to –70°C.

Additional notes:

• If the product datasheet specifies a different buffer (e.g., sterile water or a specific diluent), follow those instructions.
• For cell culture, always use sterile, endotoxin-free reagents and work under aseptic conditions.
• For working concentrations, further dilute the stock solution in cell culture medium immediately before use.
• Avoid multiple freeze-thaw cycles by aliquoting appropriately.

Summary Table:

This protocol ensures protein stability and bioactivity for cell-based assays. Always consult the specific product datasheet for any manufacturer-specific recommendations.