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Immunoassays | Inflammation & Cytokines

Erenna® IL-17A/F Heterodimer Immunoassay Kit (Cat# 03-0073-xx)

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The Erenna® IL-17A/F Heterodimer Immunoassay Kit uses a quantitative fluorescent sandwich immunoassay technique to measure IL-17A/F Heterodimer in plasma and serum samples. A capture antibody specific for IL-17A/F Heterodimer has been pre-coated onto paramagnetic microparticles (MP). The user pipettes MP, standards, and samples into uncoated microplate wells. During incubation, the IL-17A/F Heterodimer present in the sample binds to the capture antibody on the coated MP. 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-17A/F Heterodimer that has been captured onto the MP. 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-17A/F Heterodimer present in the sample when captured. The amount of IL-17A/F Heterodimer in unknown samples is interpolated from a standard curve.

Assay Performance

TABLE 1. Analytical sensitivity of the Erenna® IL-17A/F Heterodimer Immunoassay Kit1
Lower Limit of Detection 0.023 pg/mL
Lower Limit of Quantification2 0.3 pg/mL
Upper Limit of Quantification 75 pg/mL
Low-end CV% Range 1 - 9%
Low-end CV% Average 5%
Recommended Sample Volume 100 μL
Minimum Sample Volume Required3 100 μL

1 See product insert for updated values
2 LLoQ = 20% CV and ± 20% recovery
3 based upon median [IL-17A/F Heterodimer] in a healthy reference

FIGURE 1. [IL-17A/F] in serum from 20 healthy donors (8 samples < LLoQ), with median and interquartile range.

FIGURE 2. The Erenna® IL-17A/F Heterodimer Immunoassay 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

The IL-17 family is comprised of cytokines crucial for autoimmune and inflammatory responses. Out of its 6 founding members, IL-17A and IL-17F share the greatest sequence homology with four highly conserved cysteine residues. These conserved residues form disulfide bonds that contribute to the 3- dimensional molecular structure, leading to both homodimeric IL-17A/A and IL-17F/F, and heterodimeric IL-17A/F forms. The IL-17 family has been linked to rheumatoid arthritis (RA), asthma, allergic responses, lupus, allograft rejection and anti-tumor immunity. In vitro and in vivo, IL-17A/F has been shown to be more potent than IL-17F/F but less potent than IL-17A/A in inducing pro-inflammatory activities.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Interleukin-17A/F Heterodimer
UniProtKB/Swiss-Prot ID:
Protein-protein interaction database:
Alternative Names:
Gene Names:
Synonym Gene Names:
Biological Processes: apoptosis, cell-cell signaling, immune response, inflammatory response, positive regulation of transcription from RNA polymerase II promoter, protein amino acid glycosylation
Molecular Function: cytokine activity

* Source Info from www.uniprot.org.

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