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Solid-phase immunoassay for human immunoglobulin E: use of /sup 125/I-labeled protein A as a tracer. [/sup 125/I tracer technique]

Journal Article · · Anal. Biochem.; (United States)

A solid-phase immunoassay has been developed for human immunoglobulin (Ig) E. The specific binding of /sup 125/I-labeled protein A (/sup 125/I-PA) to the Fc region of rabbit IgG anti-IgE served as a quantitative measure of specific anti-IgE antibody bound to the IgE beads under optimal assay conditions. Inhibition of antibody binding by known amounts of standard IgE was reflected in a decreased binding of /sup 125/I-PA. The degree of inhibition of /sup 125/I-PA binding was related to the amount of fluid-phase IgE present and gave a standard curve which was used to determine the concentration of IgE in test samples. The sensitivity of this method and a double antibody radioimmunoassay (RIA), which was developed using the same IgE preparation and anti-IgE antibody, was approximately the same. These assays gave similar results when used to determine levels of IgE in normal human sera that had been absorbed with protein A-Sepharose to remove components responsible for specific and nonspecific interference in the assays.

Research Organization:
National Cancer Inst., Bethesda, MD
OSTI ID:
6133374
Journal Information:
Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 93:1; ISSN ANBCA
Country of Publication:
United States
Language:
English