Capture assay for specific IgE. An improved quantitative method.
A capture assay for the measurement of specific IgE in the serum of allergic patients is described, using monoclonal anti-human IgE (coated to the wells of a microtiter plate) and biotinylated allergens in solution. In a single incubation, IgE is bound to the solid phase through the Fc fragment and biotinylated allergens react with their specific IgE Fab regions, if present. In a second step, streptavidin-HRP conjugate is added to reveal the amount of biotin fixed on the solid phase. Quantitative determinations are obtained by comparison with a standard curve of total IgE incubated with a biotinylated monoclonal anti-IgE, complementary to the one employed as capture antibody. The assay is unaffected by allergen-specific IgG, shows good intra- and interassay reproducibility and is linear over a wide range of specific IgE concentrations. The method could be used with a wide range of different allergens.
Olivieri
V
V
Radim S.p.A., R.&D. Laboratories, (Rome), Italy.
Beccarini
I
I
Gallucci
G
G
Romano
T
T
Santoro
F
F
D000818:Animals; D000918:Antibody Specificity; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007073:Immunoglobulin E / Q000097:blood*; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D011852:Radioallergosorbent Test; D015203:Reproducibility of Results
D007074:Immunoglobulin G; D007073:Immunoglobulin E