Development of a sensitive enzyme immunoassay for the detection of phenyl-β-o-thioglucuronide in human urine

Norbert Staimer, Shirley J. Gee, Bruce D. Hammock

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Immunoassays for the measurement of glucuronides in human urine can be a helpful tool for the assessment of human exposure to toxic chemicals. Therefore an enzyme immunoassay (EIA) for the specific detection of phenyl-β-D-thioglucuronide was developed. The immunoconjugate was formed by coupling p-aminophenyl-β-D-thioglucuronide to the carrier protein thyroglobulin leaving an exposed glucuronic acid. The hapten-protein conjugate was adsorbed to gold colloids in order to enhance the immunogenic effect. Rabbits were injected with the immunogold conjugates to raise polyclonal antibodies. The resulting competitive assay showed an inhibition by phenyl-β-D-thioglucuronide at sample concentrations of 23.0 ± 1.3 ng/mL (50% B/B0) and a high cross-reactivity to p-aminophenyl-β-D-thioglucuronide (120%). Little cross-reactivities (< 2%) were observed for potential urinary cross reactants. In addition human urine samples were incubated with β-glucuronidase in order to investigate the EIA for specific matrix effects. An integration of high-performance liquid chromatography (HPLC) and EIA was developed in an attempt to decrease the matrix effects and increase the sensitivity of the overall method. The hyphenated technique HPEC-EIA may be used to monitor human exposure to toxic thiophenol which is excreted by mammals as urinary phenyl thioglucuronide.

Original languageEnglish (US)
Pages (from-to)273-279
Number of pages7
JournalFresenius' Journal of Analytical Chemistry
Volume369
Issue number3-4
StatePublished - 2001

Fingerprint

Immunoenzyme Techniques
Urine
Poisons
Enzymes
Immunoconjugates
Gold Colloid
Glucuronic Acid
Mammals
Haptens
Thyroglobulin
Glucuronidase
Glucuronides
High performance liquid chromatography
Immunoassay
Assays
Carrier Proteins
High Pressure Liquid Chromatography
Rabbits
Antibodies
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Clinical Biochemistry

Cite this

Development of a sensitive enzyme immunoassay for the detection of phenyl-β-o-thioglucuronide in human urine. / Staimer, Norbert; Gee, Shirley J.; Hammock, Bruce D.

In: Fresenius' Journal of Analytical Chemistry, Vol. 369, No. 3-4, 2001, p. 273-279.

Research output: Contribution to journalArticle

Staimer, Norbert ; Gee, Shirley J. ; Hammock, Bruce D. / Development of a sensitive enzyme immunoassay for the detection of phenyl-β-o-thioglucuronide in human urine. In: Fresenius' Journal of Analytical Chemistry. 2001 ; Vol. 369, No. 3-4. pp. 273-279.
@article{62a3999d78304d868c8127f1c259c146,
title = "Development of a sensitive enzyme immunoassay for the detection of phenyl-β-o-thioglucuronide in human urine",
abstract = "Immunoassays for the measurement of glucuronides in human urine can be a helpful tool for the assessment of human exposure to toxic chemicals. Therefore an enzyme immunoassay (EIA) for the specific detection of phenyl-β-D-thioglucuronide was developed. The immunoconjugate was formed by coupling p-aminophenyl-β-D-thioglucuronide to the carrier protein thyroglobulin leaving an exposed glucuronic acid. The hapten-protein conjugate was adsorbed to gold colloids in order to enhance the immunogenic effect. Rabbits were injected with the immunogold conjugates to raise polyclonal antibodies. The resulting competitive assay showed an inhibition by phenyl-β-D-thioglucuronide at sample concentrations of 23.0 ± 1.3 ng/mL (50{\%} B/B0) and a high cross-reactivity to p-aminophenyl-β-D-thioglucuronide (120{\%}). Little cross-reactivities (< 2{\%}) were observed for potential urinary cross reactants. In addition human urine samples were incubated with β-glucuronidase in order to investigate the EIA for specific matrix effects. An integration of high-performance liquid chromatography (HPLC) and EIA was developed in an attempt to decrease the matrix effects and increase the sensitivity of the overall method. The hyphenated technique HPEC-EIA may be used to monitor human exposure to toxic thiophenol which is excreted by mammals as urinary phenyl thioglucuronide.",
author = "Norbert Staimer and Gee, {Shirley J.} and Hammock, {Bruce D.}",
year = "2001",
language = "English (US)",
volume = "369",
pages = "273--279",
journal = "Fresenius Zeitschrift fur Analytische Chemie",
issn = "0016-1152",
publisher = "Springer Verlag",
number = "3-4",

}

TY - JOUR

T1 - Development of a sensitive enzyme immunoassay for the detection of phenyl-β-o-thioglucuronide in human urine

AU - Staimer, Norbert

AU - Gee, Shirley J.

AU - Hammock, Bruce D.

PY - 2001

Y1 - 2001

N2 - Immunoassays for the measurement of glucuronides in human urine can be a helpful tool for the assessment of human exposure to toxic chemicals. Therefore an enzyme immunoassay (EIA) for the specific detection of phenyl-β-D-thioglucuronide was developed. The immunoconjugate was formed by coupling p-aminophenyl-β-D-thioglucuronide to the carrier protein thyroglobulin leaving an exposed glucuronic acid. The hapten-protein conjugate was adsorbed to gold colloids in order to enhance the immunogenic effect. Rabbits were injected with the immunogold conjugates to raise polyclonal antibodies. The resulting competitive assay showed an inhibition by phenyl-β-D-thioglucuronide at sample concentrations of 23.0 ± 1.3 ng/mL (50% B/B0) and a high cross-reactivity to p-aminophenyl-β-D-thioglucuronide (120%). Little cross-reactivities (< 2%) were observed for potential urinary cross reactants. In addition human urine samples were incubated with β-glucuronidase in order to investigate the EIA for specific matrix effects. An integration of high-performance liquid chromatography (HPLC) and EIA was developed in an attempt to decrease the matrix effects and increase the sensitivity of the overall method. The hyphenated technique HPEC-EIA may be used to monitor human exposure to toxic thiophenol which is excreted by mammals as urinary phenyl thioglucuronide.

AB - Immunoassays for the measurement of glucuronides in human urine can be a helpful tool for the assessment of human exposure to toxic chemicals. Therefore an enzyme immunoassay (EIA) for the specific detection of phenyl-β-D-thioglucuronide was developed. The immunoconjugate was formed by coupling p-aminophenyl-β-D-thioglucuronide to the carrier protein thyroglobulin leaving an exposed glucuronic acid. The hapten-protein conjugate was adsorbed to gold colloids in order to enhance the immunogenic effect. Rabbits were injected with the immunogold conjugates to raise polyclonal antibodies. The resulting competitive assay showed an inhibition by phenyl-β-D-thioglucuronide at sample concentrations of 23.0 ± 1.3 ng/mL (50% B/B0) and a high cross-reactivity to p-aminophenyl-β-D-thioglucuronide (120%). Little cross-reactivities (< 2%) were observed for potential urinary cross reactants. In addition human urine samples were incubated with β-glucuronidase in order to investigate the EIA for specific matrix effects. An integration of high-performance liquid chromatography (HPLC) and EIA was developed in an attempt to decrease the matrix effects and increase the sensitivity of the overall method. The hyphenated technique HPEC-EIA may be used to monitor human exposure to toxic thiophenol which is excreted by mammals as urinary phenyl thioglucuronide.

UR - http://www.scopus.com/inward/record.url?scp=0034935568&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034935568&partnerID=8YFLogxK

M3 - Article

C2 - 11293704

AN - SCOPUS:0034935568

VL - 369

SP - 273

EP - 279

JO - Fresenius Zeitschrift fur Analytische Chemie

JF - Fresenius Zeitschrift fur Analytische Chemie

SN - 0016-1152

IS - 3-4

ER -