An enzymeimmunoassay for total thyroxine using avidin-biotin separation system and thyroxine-peroxidase conjugate

An avidin-biotin enzymeimmunoassay for total thyroxine in serum is described. Avidin was adsorbed to biotinylated bovine serum albumin coated tubes prepared with glutaraldehyde as coupling agent. In the enzymeimmunoassay, affinity purified biotinylated anti-thyroxine IgG, sample or standards, and th...

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Bibliographic Details
Published inJournal of immunoassay (Monticello, N.Y.) Vol. 16; no. 3; p. 325
Main Authors Akman, S, Karaca, L, Kutluay, T
Format Journal Article
LanguageEnglish
Published United States 01.08.1995
Subjects
Animals
Avidin - standards
Biotin - standards
Chromatography, Affinity
Horseradish Peroxidase
Humans
Immune Sera - chemistry
Immunoenzyme Techniques - standards
Immunoglobulin G - chemistry
Peroxidases - immunology
Sheep
Thyroxine - blood
Thyroxine - immunology
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Summary:An avidin-biotin enzymeimmunoassay for total thyroxine in serum is described. Avidin was adsorbed to biotinylated bovine serum albumin coated tubes prepared with glutaraldehyde as coupling agent. In the enzymeimmunoassay, affinity purified biotinylated anti-thyroxine IgG, sample or standards, and thyroxine-horseradish peroxidase were simultaneously added to the avidin coated tubes. The bound enzymatic activity was then measured with o-phenylenediamine and H2O2. Results showed that the assay has good precision (within-assay CV% less than 10% and between assay 11.7% in hypo- and 6.9% in hyperthyroid range), good assay range (0-800 nmol/L), good sensitivity (4 nmol/L), and can be performed in 2.5 hours. The results obtained correlate well (r = 0.93) with those of an RIA.
ISSN:0197-1522
2332-4090
DOI:10.1080/15321819508013566