1. 23. Clinical laboratory assessment of IgE-dependent hypersensitivity
Robert G. Hamilton, PhD, DABMLI, N.Franklin Adkinson, MD 
Journal of Allergy and Clinical Immunology 
Volume 111, Issue 2, Pages S687-S701 (February 2003)
DOI: /mai
Copyright © 2003 Mosby, Inc. Terms and Conditions

2. Fig. 1
A, Schematic of the steps involved in the allergosorbent test. For diagnostic purposes (IgE antibody detection, allergen-specific antibody in serum of different isotypes (IgE, IgG, IgA) is bound to optimal amounts of allergen (closed circles) insolubilized onto a solid phase. After removal of unbound serum proteins with buffer, bound IgE antibody is detected with 125I or enzyme-labeled antihuman IgE detection antibody in the RAST or enzyme-linked allergosorbent test (EAST), respectively. The response (counts per minute [CPM] bound to the allergosorbent or residual optical density) after a last buffer wash is proportional to the amount of specific IgE antibody in the original serum. Multiple dilutions of reference and test sera are simultaneously analyzed, and their dose-response curves should dilute out in parallel.
FIG 1. B, For allergen potency assessment, the EAST/RAST inhibition assay is used. In these assays, increasing amounts of fluid-phase allergen are added to a fixed amount of IgE antibody-containing serum before the allergosorbent is added. As the potency of the fluid-phase allergen increases, there is less free IgE available to bind the allergosorbent. Thus, there is proportionally less labeled anti-human IgE bound and a lower response signal. This gives rise to a negative dose-response curve.
Journal of Allergy and Clinical Immunology  , S687-S701DOI: ( /mai )
Copyright © 2003 Mosby, Inc. Terms and Conditions

3. Fig. 1
A, Schematic of the steps involved in the allergosorbent test. For diagnostic purposes (IgE antibody detection, allergen-specific antibody in serum of different isotypes (IgE, IgG, IgA) is bound to optimal amounts of allergen (closed circles) insolubilized onto a solid phase. After removal of unbound serum proteins with buffer, bound IgE antibody is detected with 125I or enzyme-labeled antihuman IgE detection antibody in the RAST or enzyme-linked allergosorbent test (EAST), respectively. The response (counts per minute [CPM] bound to the allergosorbent or residual optical density) after a last buffer wash is proportional to the amount of specific IgE antibody in the original serum. Multiple dilutions of reference and test sera are simultaneously analyzed, and their dose-response curves should dilute out in parallel.
FIG 1. B, For allergen potency assessment, the EAST/RAST inhibition assay is used. In these assays, increasing amounts of fluid-phase allergen are added to a fixed amount of IgE antibody-containing serum before the allergosorbent is added. As the potency of the fluid-phase allergen increases, there is less free IgE available to bind the allergosorbent. Thus, there is proportionally less labeled anti-human IgE bound and a lower response signal. This gives rise to a negative dose-response curve.
Journal of Allergy and Clinical Immunology  , S687-S701DOI: ( /mai )
Copyright © 2003 Mosby, Inc. Terms and Conditions