High-throughput automated luminescent magnetic particle-based immunoassay to monitor human exposure to pyrethroid insecticides

Chang Ahn Ki, Pete Lohstroh, Shirley J. Gee, Nancy A. Gee, Bill Lasley, Bruce D. Hammock

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

We have developed a sensitive, automated, competitive chemiluminescent immunoassay for the detection of 3-phenoxybenzoic acid (3-PBA), a metabolite common to many pyrethroid insecticides. The system uses a competitive hapten-protein conjugate that has been labeled with an acridinium ester as the chemiluminescent probe and secondary antibody-coated paramagnetic particles for the separation. After the immunoassay reagents and samples are combined for the competitive incubation step, a fully automated system is used to load the postincubation mixture into a delivery cuvette, facilitating the subsequent magnetic separation of the immunocomplex and the measurement of chemiluminescent signal for quantification. The immunoassay format described here supports the requirement for high throughput necessary for monitoring large numbers of samples in population-based studies. The optimized immunoassay was more sensitive than the conventional enzyme immunoassay in buffer (IC50 = 0.1 and 2 μg/L, respectively). The mixed-mode solid-phase extraction used for sample preparation to reduce possible urinary matrix effects allowed the accurate measurement of 3-PBA levels as low as 1 μg/L. The automated chemiluminescent immunoassay described here is sensitive, simple to use, and more rapid than the previously reported standard microplate assay.

Original languageEnglish (US)
Pages (from-to)8883-8890
Number of pages8
JournalAnalytical Chemistry
Volume79
Issue number23
DOIs
StatePublished - Dec 1 2007

Fingerprint

Pyrethrins
Insecticides
Throughput
Magnetic separation
Haptens
Metabolites
Assays
Buffers
Esters
Antibodies
Monitoring
Enzymes
Proteins
3-phenoxybenzoic acid

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

High-throughput automated luminescent magnetic particle-based immunoassay to monitor human exposure to pyrethroid insecticides. / Ki, Chang Ahn; Lohstroh, Pete; Gee, Shirley J.; Gee, Nancy A.; Lasley, Bill; Hammock, Bruce D.

In: Analytical Chemistry, Vol. 79, No. 23, 01.12.2007, p. 8883-8890.

Research output: Contribution to journalArticle

Ki, Chang Ahn ; Lohstroh, Pete ; Gee, Shirley J. ; Gee, Nancy A. ; Lasley, Bill ; Hammock, Bruce D. / High-throughput automated luminescent magnetic particle-based immunoassay to monitor human exposure to pyrethroid insecticides. In: Analytical Chemistry. 2007 ; Vol. 79, No. 23. pp. 8883-8890.
@article{87aca2f607854c0f84e57438da6bdfc0,
title = "High-throughput automated luminescent magnetic particle-based immunoassay to monitor human exposure to pyrethroid insecticides",
abstract = "We have developed a sensitive, automated, competitive chemiluminescent immunoassay for the detection of 3-phenoxybenzoic acid (3-PBA), a metabolite common to many pyrethroid insecticides. The system uses a competitive hapten-protein conjugate that has been labeled with an acridinium ester as the chemiluminescent probe and secondary antibody-coated paramagnetic particles for the separation. After the immunoassay reagents and samples are combined for the competitive incubation step, a fully automated system is used to load the postincubation mixture into a delivery cuvette, facilitating the subsequent magnetic separation of the immunocomplex and the measurement of chemiluminescent signal for quantification. The immunoassay format described here supports the requirement for high throughput necessary for monitoring large numbers of samples in population-based studies. The optimized immunoassay was more sensitive than the conventional enzyme immunoassay in buffer (IC50 = 0.1 and 2 μg/L, respectively). The mixed-mode solid-phase extraction used for sample preparation to reduce possible urinary matrix effects allowed the accurate measurement of 3-PBA levels as low as 1 μg/L. The automated chemiluminescent immunoassay described here is sensitive, simple to use, and more rapid than the previously reported standard microplate assay.",
author = "Ki, {Chang Ahn} and Pete Lohstroh and Gee, {Shirley J.} and Gee, {Nancy A.} and Bill Lasley and Hammock, {Bruce D.}",
year = "2007",
month = "12",
day = "1",
doi = "10.1021/ac070675l",
language = "English (US)",
volume = "79",
pages = "8883--8890",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "23",

}

TY - JOUR

T1 - High-throughput automated luminescent magnetic particle-based immunoassay to monitor human exposure to pyrethroid insecticides

AU - Ki, Chang Ahn

AU - Lohstroh, Pete

AU - Gee, Shirley J.

AU - Gee, Nancy A.

AU - Lasley, Bill

AU - Hammock, Bruce D.

PY - 2007/12/1

Y1 - 2007/12/1

N2 - We have developed a sensitive, automated, competitive chemiluminescent immunoassay for the detection of 3-phenoxybenzoic acid (3-PBA), a metabolite common to many pyrethroid insecticides. The system uses a competitive hapten-protein conjugate that has been labeled with an acridinium ester as the chemiluminescent probe and secondary antibody-coated paramagnetic particles for the separation. After the immunoassay reagents and samples are combined for the competitive incubation step, a fully automated system is used to load the postincubation mixture into a delivery cuvette, facilitating the subsequent magnetic separation of the immunocomplex and the measurement of chemiluminescent signal for quantification. The immunoassay format described here supports the requirement for high throughput necessary for monitoring large numbers of samples in population-based studies. The optimized immunoassay was more sensitive than the conventional enzyme immunoassay in buffer (IC50 = 0.1 and 2 μg/L, respectively). The mixed-mode solid-phase extraction used for sample preparation to reduce possible urinary matrix effects allowed the accurate measurement of 3-PBA levels as low as 1 μg/L. The automated chemiluminescent immunoassay described here is sensitive, simple to use, and more rapid than the previously reported standard microplate assay.

AB - We have developed a sensitive, automated, competitive chemiluminescent immunoassay for the detection of 3-phenoxybenzoic acid (3-PBA), a metabolite common to many pyrethroid insecticides. The system uses a competitive hapten-protein conjugate that has been labeled with an acridinium ester as the chemiluminescent probe and secondary antibody-coated paramagnetic particles for the separation. After the immunoassay reagents and samples are combined for the competitive incubation step, a fully automated system is used to load the postincubation mixture into a delivery cuvette, facilitating the subsequent magnetic separation of the immunocomplex and the measurement of chemiluminescent signal for quantification. The immunoassay format described here supports the requirement for high throughput necessary for monitoring large numbers of samples in population-based studies. The optimized immunoassay was more sensitive than the conventional enzyme immunoassay in buffer (IC50 = 0.1 and 2 μg/L, respectively). The mixed-mode solid-phase extraction used for sample preparation to reduce possible urinary matrix effects allowed the accurate measurement of 3-PBA levels as low as 1 μg/L. The automated chemiluminescent immunoassay described here is sensitive, simple to use, and more rapid than the previously reported standard microplate assay.

UR - http://www.scopus.com/inward/record.url?scp=36849044717&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=36849044717&partnerID=8YFLogxK

U2 - 10.1021/ac070675l

DO - 10.1021/ac070675l

M3 - Article

C2 - 17983205

AN - SCOPUS:36849044717

VL - 79

SP - 8883

EP - 8890

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 23

ER -