IgE receptors expressed on antigen presenting cells have been shown to facilitate the presentation of allergens in the presence of specific IgE, resulting in effective T cell activation at low allergen concentrations. It has been demonstrated with this assay that formation of allergen-IgE complexes increases with IgE clonality and allergen affinity. Interference with these IgE-dependent mechanisms by ‘blocking’ IgG antibodies may down regulate T cell responses and manifest as a reduction in allergic responses and long-term clinical tolerance.
The IgE-FAB assay represents an in vitro model of facilitated allergen presentation. Allergen-IgE complexes are incubated with an EBV-transformed B cell line and complexes bound to CD23 on the surface of cells are detected by flow cytometry.
Principle of the FAB-assay
- Allergen-serum complex binding
- Incubation of antibody/allergen complex with EBV-transformed B cells
- Bounded complexes to CD23 on cell surface are stained with anti-IgE
- Analyze IgE levels by flow cytometry
KEY APPLICATIONS:
This assay is suitable for monitoring inhibitory antibody function from patients receiving allergen-specific immunotherapy.
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