Citation

Wilson R. Anal. Chim. Acta 1992; 269(2): 301-306.

Copyright

(Copyright © 1992, Elsevier Publishing)

DOI

10.1016/0003-2670(92)85416-4

PMID

unavailable

Abstract

METHODS for removing contaminating FAD and alkaline phosphatase from apo-aminoacid oxidase are described. A method for phosphorylating flavin adenine dinucleotide is also described. Its hydrolysis to flavin adenine dinucleotide catalysed by alkaline phosphatase is demonstrated by liquid chromatography. Flavin adenine dinucleotide can reactivate apo-aminoacid oxidase and result in the formation of hydrogen peroxide. The hydrogen peroxide can be detected colorimetrically. These reactions are used in an enzyme amplified immunoassay for thyroid stimulating hormone in which alkaline phosphatase is the enzyme label. A limit of detection of 0.03 μIU ml-1 for thyroid stimulating hormone makes the assay suitable for diagnosing hyperthyroidism. © 1992.

Language: en

Keywords

human; article; thyrotropin; spectrophotometry; hyperthyroidism; Immunoassay; Suicide substrate; enzyme purification; enzyme immunoassay; Enzyme amplified immunoassay; Flavin adenine dinucleotide phosphate; Microtitre plate; Nitroethane; Thyroid stimulating hormone