Development of a homogeneous phosphorescent immunoassay for the detection of polychlorinated dibenzo-p-dioxins

E. G. Matveeva; E. V. Gribkova; J. R. Sanborn; S. J. Gee; B. D. Hammock; A. P. Savitsky

doi:10.1081/AL-100107297

Development of a homogeneous phosphorescent immunoassay for the detection of polychlorinated dibenzo-p-dioxins

E. G. Matveeva, E. V. Gribkova, J. R. Sanborn, S. J. Gee, B. D. Hammock, A. P. Savitsky

Entomology

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7 Scopus citations

Abstract

A homogeneous immunoassay for dioxins was developed using a phosphorescent label for detection. A dioxin derivative was conjugated to Pt-coproporphyrin. In the assay, when the antibodies against dioxin (DD3) were bound to the phosphorescent conjugate, the signal from Pt-coproporphyrin was quenched. The interaction between antibody and the conjugate was studied by time-resolved luminescence spectroscopy. As concentrations of dioxin standards increased from 39 pg/well to 2.5 ng/well the lifetime of the phosphorescence of the shortlived component increased from 25.6 microseconds to 58.9 microseconds. This increase in half-life was associated with a dose dependent quenching of the phosphorescence. The inhibition obtained is similar to that for a reported enzyme-based immunoassay, but the data were obtained in minutes instead of hours.

Original language	English (US)
Pages (from-to)	2311-2320
Number of pages	10
Journal	Analytical Letters
Volume	34
Issue number	13
DOIs	https://doi.org/10.1081/AL-100107297
State	Published - 2001

Keywords

Homogeneous immunoassay
Phosphorescence
Polychlorinated dibenzo-p-dioxins
Pt-coproporphyrin
Time-resolved luminescence spectroscopy

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Biochemistry
Analytical Chemistry

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title = "Development of a homogeneous phosphorescent immunoassay for the detection of polychlorinated dibenzo-p-dioxins",

abstract = "A homogeneous immunoassay for dioxins was developed using a phosphorescent label for detection. A dioxin derivative was conjugated to Pt-coproporphyrin. In the assay, when the antibodies against dioxin (DD3) were bound to the phosphorescent conjugate, the signal from Pt-coproporphyrin was quenched. The interaction between antibody and the conjugate was studied by time-resolved luminescence spectroscopy. As concentrations of dioxin standards increased from 39 pg/well to 2.5 ng/well the lifetime of the phosphorescence of the shortlived component increased from 25.6 microseconds to 58.9 microseconds. This increase in half-life was associated with a dose dependent quenching of the phosphorescence. The inhibition obtained is similar to that for a reported enzyme-based immunoassay, but the data were obtained in minutes instead of hours.",

keywords = "Homogeneous immunoassay, Phosphorescence, Polychlorinated dibenzo-p-dioxins, Pt-coproporphyrin, Time-resolved luminescence spectroscopy",

author = "Matveeva, {E. G.} and Gribkova, {E. V.} and Sanborn, {J. R.} and Gee, {S. J.} and Hammock, {B. D.} and Savitsky, {A. P.}",

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T1 - Development of a homogeneous phosphorescent immunoassay for the detection of polychlorinated dibenzo-p-dioxins

AU - Matveeva, E. G.

AU - Gribkova, E. V.

AU - Sanborn, J. R.

AU - Gee, S. J.

AU - Hammock, B. D.

AU - Savitsky, A. P.

PY - 2001

Y1 - 2001

N2 - A homogeneous immunoassay for dioxins was developed using a phosphorescent label for detection. A dioxin derivative was conjugated to Pt-coproporphyrin. In the assay, when the antibodies against dioxin (DD3) were bound to the phosphorescent conjugate, the signal from Pt-coproporphyrin was quenched. The interaction between antibody and the conjugate was studied by time-resolved luminescence spectroscopy. As concentrations of dioxin standards increased from 39 pg/well to 2.5 ng/well the lifetime of the phosphorescence of the shortlived component increased from 25.6 microseconds to 58.9 microseconds. This increase in half-life was associated with a dose dependent quenching of the phosphorescence. The inhibition obtained is similar to that for a reported enzyme-based immunoassay, but the data were obtained in minutes instead of hours.

AB - A homogeneous immunoassay for dioxins was developed using a phosphorescent label for detection. A dioxin derivative was conjugated to Pt-coproporphyrin. In the assay, when the antibodies against dioxin (DD3) were bound to the phosphorescent conjugate, the signal from Pt-coproporphyrin was quenched. The interaction between antibody and the conjugate was studied by time-resolved luminescence spectroscopy. As concentrations of dioxin standards increased from 39 pg/well to 2.5 ng/well the lifetime of the phosphorescence of the shortlived component increased from 25.6 microseconds to 58.9 microseconds. This increase in half-life was associated with a dose dependent quenching of the phosphorescence. The inhibition obtained is similar to that for a reported enzyme-based immunoassay, but the data were obtained in minutes instead of hours.

KW - Homogeneous immunoassay

KW - Phosphorescence

KW - Polychlorinated dibenzo-p-dioxins

KW - Pt-coproporphyrin

KW - Time-resolved luminescence spectroscopy

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