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Immunoassays | Inflammation & Cytokines

Erenna® IL-15 Immunoassay Kit (Cat# 03-0058-xx)

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The Erenna® IL-15 Immunoassay uses a quantitative fluorescent sandwich immunoassay technique to measure IL-15 in human serum and plasma samples. A capture antibody specific for human IL-15 has been pre-coated onto paramagnetic microparticles (MPs). The user pipettes MPs, standards, and samples into uncoated microplate wells. During incubation, the IL-15 present in the sample binds to the capture antibody on the coated MPs. Unbound molecules are washed away during the subsequent buffer exchange and wash steps. Fluor-labeled detection antibody is added to each well and incubated. This detection antibody recognizes and binds to IL-15 that has been captured onto the MPs. During the following wash step the MPs are transferred to a clean plate. Elution buffer is then added and incubated. The elution buffer dissociates the bound protein sandwich from the MP surface, releasing the labeled antibodies. These antibodies are separated during transfer to a final microplate. The plate is loaded into the Erenna® System where the labeled molecules are detected and counted. The number of fluor-labeled detection antibodies counted is directly proportional to the amount of IL-15 present in the sample when captured. The amount of IL-15 in unknown samples is interpolated from a standard curve.

Assay Performance

TABLE 1. Analytical sensitivity of the Erenna® IL-15 Immunoassay Kit1
Lower Limit of Detection 0.01 pg/mL
Lower Limit of Quantification2 0.1 pg/mL
Upper Limit of Quantification 25 pg/mL
Low-end CV% Range 7 - 16%
Low-end CV% Average 10%
Recommended Sample Volume 100 μL
Minimum Sample Volume Required3 3 μL

1 See product insert for updated values
2 LLoQ = 20% CV and ± 20% recovery
3 based upon median [IL-15] in a healthy reference

FIGURE 1. [IL-15] in EDTA plasma from 10 healthy donors, with median and interquartile range. The Erenna® IL-15 Immunoassay Evaluation Reagent Kit reliably quantifies IL-15 in healthy subjects, who have a median [IL-15] of 3.38 pg/mL that is well above the detection limit of 0.1 pg/mL.

FIGURE 2. Erenna® IL-15 Immunoassay Evaluation Reagent Kit low-end standard curve signal (left) and curve fit (right).

Representative data shown for demonstration purposes only. Individual results may vary depending upon samples tested and protocol used.

Biology and Disease

Interleukin-15 (IL-15) is a cytokine that bears structural and biological similarities to IL-2, as both cytokines are triggered by the presence of exogenous bacteria and viruses. Unlike IL-2, which is expressed mainly by activated monocytes, IL-15 is constitutively expressed by a variety of tissues and cell types, including macrophages, dendritic cells, osteoclasts and fibroblasts. IL-15 is known to affect the proliferation of T-, B- and natural killer cells, and initiates the up-regulation of the cytokines GM-CSF and TNFα. The pro-inflammatory properties of IL-15 elicit autoimmune activity and elevated levels of IL-15 have been associated with psoriasis, multiple sclerosis, rheumatoid arthritis and systemic lupus.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Interleukin-15
UniProtKB/Swiss-Prot ID: P40933
Protein-protein interaction database: P40933
Alternative Names:
Gene Names: IL15
Synonym Gene Names:
Function: Cytokine that stimulates the proliferation of T-lymphocytes. Stimulation by IL-15 requires interaction of IL-15 with components of IL-2R, including IL-2R beta and probably IL-2R gamma but not IL-2R alpha.
Biological Processes: immune response, cell-cell signaling
Molecular Function: cytokine activity, cytokine receptor binding, signal transducer activity

* Source Info from www.uniprot.org.

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