Enhanced chemiluminescent assays for horseradish peroxidase and their application in immunoassays

Thorpe, Gary Harold Gregory Henry (1986). Enhanced chemiluminescent assays for horseradish peroxidase and their application in immunoassays. University of Birmingham. Ph.D.

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Abstract

Certain 6-hydroxybenzothiazole and phenol derivatives enhance light emission from the horseradish peroxidase catalysed oxidation of cyclic diacyl hydrazides such as luminol. These reactions produce novel, enhanced chemiluminescent assays for horseradish peroxidase labels used in immunoassays. Enhanced assays are rapid and sensitive, light emission is relatively prolonged, decays slowly, and the light intensity can be a 1000- fold that of unenhanced reactions. The reactions have been partially characterised and their emission spectra shown to be remarkably similar to unenhanced reactions. Two hypotheses are proposed to explain the mechanism of enhancement.

The utility of enhanced chemiluminescent assays for horseradish peroxidase conjugates was demonstrated in a range of immunoassays, both immunoextraction and competitive, in conjunction with a variety of solid supports such as plastic beads, tubes and microtitre plates. The rapid and simply performed enhanced chemiluminescent assays have been used both to replace conventional colorimetric end-points, and in the development of rapid immunoassays. No interferences were encountered with clinical samples in enhanced chemiluminescently monitored immunoassays and results correlated well with those determinedcolorimetrically . Characteristics of enhanced reactions enable simplification of the design of instrumentation required for routine immunoassays, and allow quantitation of light emission using either photomultiplier tubes or instant photographic film. Such systems should facilitate the introduction of chemiluminescently monitored immunoassays into clinical laboratories.

Type of Work:	Thesis (Doctorates > Ph.D.)
Award Type:	Doctorates > Ph.D.
Licence:	All rights reserved
College/Faculty:	Faculties (to 1997) > Faculty of Science
School or Department:	Department of Clinical Chemistry
Funders:	None/not applicable
Subjects:	Q Science > QD Chemistry
URI:	http://etheses.bham.ac.uk/id/eprint/13218

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