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Immunoassays | Inflammation & Cytokines

Erenna® IL-22 Immunoassay Kit (Cat# 03-0059-xx)

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Introduction

The Erenna® IL-22 Immunoassay Kit uses a quantitative fluorescent sandwich immunoassay technique to measure IL-22 in plasma and serum samples. A capture antibody specific for IL-22 has been pre-coated onto paramagnetic microparticles (MP). The user pipettes MP, standards, and samples into uncoated microplate wells. During incubation, the IL-22 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-22 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-22 present in the sample when captured. The amount of IL-22 in unknown samples is interpolated from a standard curve.

Assay Performance


TABLE 1. Analytical sensitivity of the Erenna® IL-22 Immunoassay Kit1
Lower Limit of Detection 0.024 pg/mL
Lower Limit of Quantification2 0.2 pg/mL
Upper Limit of Quantification 100 pg/mL
Low-end CV% Range 2 - 20%
Low-end CV% Average 11%
Recommended Sample Volume 100 μL
Minimum Sample Volume Required3 10 μL

1 See product insert for updated values
2 LLoQ = 20% CV and ± 20% recovery
3 based upon median [IL-22] in a healthy reference


FIGURE 1. [IL-22] in EDTA plasma from 10 healthy donors, with median and interquartile range. The Erenna® IL-22 Immunoassay Kit reliably quantifies IL-22 in healthy subjects, who have a median [IL-22] of 3.3 pg/mL that is well above the detection limit of 0.024 pg/mL.

FIGURE 2. Erenna® IL-22 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

TInterleukin-22 (IL-22), also known as IL-10-related T-cellderived inducible factor (IL-TIF), is a member of the IL-10 cytokine family. Mainly produced by activated T-cells and natural killer cells, IL-22 binds to a receptor complex with two components, CRF2-4 and IL-22R. The signaling cascade is said to up-regulate pro-inflammatory cytokines and antimicrobial peptides, and induce production of acutephase reactants in liver and hepatoma cells. The biological function of IL-22 is associated with many inflammatory disorders, including Crohn’s disease, inflammatory bowel disease, psoriasis, rheumatoid arthritis and ulcerative colitis. Thus, IL-22 remains an attractive therapeutic target because of its pro-inflammatory properties.


TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Interleukin-22
UniProtKB/Swiss-Prot ID: Q9GZX6
Protein-protein interaction database: Q9GZX6
Alternative Names: Cytokine Zcyto18, IL-10-related T-cell-derived-inducible factor
Gene Names: IL22
Synonym Gene Names: ILTIF, ZCYTO18
Function: Cytokine that contributes to the inflammatory response in vivo.
Biological Processes: acute-phase response, cell-cell signaling
Molecular Function: cytokine activity, interleukin-22 receptor binding

* Source Info from www.uniprot.org.


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