Human IFN-gamma Autoantibody Assay ELISA Kit

Molecule:

IFN-gamma

Species:

Human

Cat. No.:

CKH022 pdf (datasheet)

Quantity/Size:

1 x 96 tests

Price:

$1,175.00 BUY

Description:

Introduction
An increasing number of reports describe the presence of naturally occurring autoantibodies to a variety of cytokines in the circulation of human individuals without any obvious adverse health effects. Dysregulation of this immune network, leading to enhanced autoantibody production have also been described, resulting in a pathogenic effect (1). For example, high levels of neutralizing anti-IFN-γ autoantibodies (IFAAs) have been described to underlie disseminated non-tuberculous mycobacterial infections (2, 3). Other anti-cytokine autoantibodies associated with disease pathogenesis have been described for GM-CSF, Interleukin 17A/F (IL-17A/F) and IL-22, which are involved in pulmonary alveolar proteinosis (PAP), chronic mucocutaneous candidiasis and autoimmune polyendocrine syndrome type I, respectively (4,5).

In order to measure IFN-γ autoantibodies, a highly sensitive immuno-enzymatic capture assay was developed for the quantitative determination of IFAAs in human serum and plasma. The assay provides an attractive tool to monitor disease progression and response to therapy. The performance of the assay is easy and straightforward and therefore simple to implement. The design of the supplied pre-coated plate is optimized to preserve the native conformation of the IFN-γ molecule and avoids steric hindrance of antibody binding. In addition, the assay includes an internal negative control for each sample analysis, thereby minimizing the chance on false positivity. Overall, the Human IFN-γ Autoantibody Assay is a highly sensitive, reproducible assay providing maximal flexibility to study IFAAs in health and disease.

Principle of the test
The kit is a sensitive antigen capture assay for autoantibody determinations in human serum and plasma samples. The assay consists of streptavidin-coated 96-well strip plates with immobilized biotin-labeled IFN-γ molecules. Control wells contain an immobilized control agent for measuring non-specific binding. Standards, controls and samples are added to the wells, and (auto)antibodies present in the diluted samples bind to the captured antigen. Next, wells are washed and incubated with an alkaline phosphatase (AP)-labeled anti-human IgG conjugate. After washing away unbound conjugate, the enzymatic activity is detected by addition of a ready-to-use p-NitroPhenyl Phosphate (pNPP) substrate. Finally, the enzymatic reaction is stopped and the optical density (OD) is read at 405 nm (reference 650 nm).

 

References

1. Browne S.K., et al., Immunodeficiency secondary to anti-cytokine autoantibodies. Curr Opin Allergy Clin Immunol, 2010. 10(6): 534-41.

2. Chi C.Y., et al., Anti-IFN-gamma autoantibodies in adults with disseminated nontuberculous mycobacterial infections are associated with HLA-DRB1*16:02 and HLA-DQB1*05:02 and the reactivation of latent varicella-zoster virus infection. Blood, 2013. 121(8): 1357-66.

3. Patel S.Y., et al., Anti-IFN-gamma autoantibodies in disseminated nontuberculous mycobacterial infections. J Immunol, 2005. 175(7): 4769-76.

4. Puel A., et al., Autoantibodies against IL-17A, IL-17F, and IL-22 in patients with chronic mucocutaneous candidiasis and autoimmune polyendocrine syndrome type I. J Exp Med, 2010. 207(2): 291-7.

5. Uchida K., et al., High-affinity autoantibodies specifically eliminate granulocyte-macrophage colonystimulating factor activity in the lungs of patients with idiopathic pulmonary alveolar proteinosis. Blood, 2004. 103(3): 1089-98.

Data PDF:

CKH022

Storage/Stability:

Store kit at 4°C

Application:

ELISA