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Immunoassays | Inflammation & Cytokines

Erenna® G-CSF Immunoassay Kit (Cat# 03-0047-xx)

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

Assay Performance

TABLE 1. Analytical sensitivity of the Erenna® G-CSF Immunoassay Kit1
Lower Limit of Detection 0.03 pg/mL
Lower Limit of Quantification2 0.08 pg/mL
Upper Limit of Quantification 20 pg/mL
Low-end CV% Range 5 - 9%
Low-end CV% Average 7%
Recommended Sample Volume 10 µL
Minimum Sample Volume Required3 1 µL

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

FIGURE 1. [G-CSF] in EDTA plasma from 10 healthy donors, with median and interquartile range. The Erenna® G-CSF Immunoassay Kit reliably quantifies G-CSF in healthy subjects, who have a median [G-CSF] of 17 pg/mL that is well above the detection limit of 0.03 pg/mL.

FIGURE 2. Erenna® G-CSF 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

Granulocyte-Colony Stimulating Factor (G-CSF) is an endogenous hematopoietic agent activated and expressed by many types of cells, including macrophages, tumor cells, fibroblasts and endothelial cells. One of the main functions of G-CSF is to regulate formation of neutrophils during hematopoiesis, which are important for proper immune function. For example, in cancer patients, reduced hematopoeisis from chemotherapy results in low neutrophil production, which jeopardizes the immune system, leaving the body susceptible to infectious attacks. G-CSF therapy has been shown to restore hematopoiesis in cancer and leukemic patients by stimulating white blood cell production. Interestingly, G-CSF has also demonstrated effectiveness in improving cardiac function following a myocardial infarction.

TABLE 2. Additional UniProtKB/Swiss-Prot Information*
Protein Name: Granulocyte colony-stimulating factor
UniProtKB/Swiss-Prot ID: P09919
Protein-protein interaction database: P09919
Alternative Names: Pluipoietin, INN=Filgrastim, INN=Lenograstim
Gene Names: CSF3
Synonym Gene Names: c17orf333, GCSF
Function: Granulocyte/macrophage colony-stimulating factors are cytokines that act in hematopoiesis by controlling the production, differentiation, and function of 2 related white cell populations of the blood, the granulocytes and the monocytes-macrophages. This CSF induces granulocytes.
Biological Processes: cytokine-mediated signaling pathway, granulocyte differentiation, immune response, positive regulation of cell proliferation
Molecular Function: cytokine activity, enzyme binding, granulocyte colony-stimulating factor receptor binding, growth factor activity

* Source Info from www.uniprot.org.

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