Erenna® IL-17F Immunoassay Kit (Cat# 03-0018-xx)

Introduction

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

Assay Performance

TABLE 1. Analytical sensitivity of the Erenna® IL-17F Immunoassay Kit¹
Lower Limit of Detection	0.5 pg/mL
Lower Limit of Quantification²	1.6 pg/mL
Upper Limit of Quantification	400 pg/mL
Low-end CV% Range	2 - 21%
Low-end CV% Average	7%
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-17F] in a healthy reference

FIGURE 1. [IL-17F] in serum from 20 healthy donors, with median and interquartile range. The Erenna® IL-17F Immunoassay Kit reliably quantifies IL-17F in serum from healthy subjects, who have a median [IL-17F] of 20.9 pg/mL that is well above the detection limit of 0.5 pg/mL.

FIGURE 2. Erenna® IL-17F 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

Interleukin-17F (IL-17F) is a member of a group of cytokines called the IL-17 family. Other members in the family include IL-17A, IL-17B, IL-17C and IL-17E (IL-17E is also known as IL-25). IL-17F is involved in mediating pro-inflammatory responses by inducing the production of many other cytokines, chemokines, and prostaglandins from many cell types. The increased expression of chemokines attracts other cells, including neutrophils. IL-17F has many physiological functions, such as airway remodeling. As a result of these roles, the IL-17 family has been linked to many immune/autoimmune related diseases, including rheumatoid arthritis (RA), asthma, allergic responses, lupus, allograft rejection and anti-tumor immunity.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name:	Interleukin-17F
UniProtKB/Swiss-Prot ID:	Q96PD4
Protein-protein interaction database:	Q96PD4
Alternative Names:	Cytokine ML-1, Interleukin-24
Gene Names:	IL17F
Synonym Gene Names:	IL24
Function:	Stimulates the production of other cytokines such as IL-6, IL-8 and granulocyte colony-stimulating factor, and can regulate cartilage matrix turnover. Stimulates PBMC and T-cell proliferation. Inhibits angiogenesis.
Biological Processes:	cartilage development, inflammatory response, lymphotoxin A biosynthetic process, negative regulation of angiogenesis, proteoglycan metabolic process, regulation of granulocyte macrophage colony-stimulating factor biosynthetic process, regulation of interleukin-2 biosynthetic process, regulation of interleukin-6 biosynthetic process, regulation of interleukin-8 biosynthetic process, regulation of transforming growth factor beta receptor signaling pathway
Molecular Function:	cytokine activity, cytokine binding, protein homodimerization activity

* Source Info from www.uniprot.org.

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