Hapten and antibody production for a sensitive immunoassay determining a human urinary metabolite of the pyrethroid insecticide permethrin

Ki Chang Ahn, Takaho Watanabe, Shirley J. Gee, Bruce D. Hammock

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Permethrin is the most popular synthetic pyrethroid insecticide in agriculture and public health. For the development of the enzyme-linked immunosorbent assay (ELISA) to evaluate human exposure to permethrin, the glycine conjugate (DCCA-glycine) of a major metabolite, cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (DCCA), of permethrin was established as the target analyte. Four different types of the cis- and trans-isomers of immunizing haptens were synthesized as follows: N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine (hapten 3), N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1- carbonyl)-4-amino-L-phenyl-alanine (hapten 5), N-(N-(cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine)-amino-6-(2, 4-dinitrophenyl)aminohexanoic acid (hapten 9), and N-(cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine-4-oxobutanoic acid (hapten 24). Sixteen polyclonal antibodies produced against each cis- or trans-hapten-thyroglobulin conjugate as immunogens were screened against numerous hapten-bovine serum albumin conjugates as coating antigens. Six ELISAs with both a heterologous hapten structure and a heterologous hapten configuration (cis/trans or trans/cis) between antibody and coating antigen showed a high sensitivity for the target analyte. The IC50 was 1.3, 2.1, and 2.2 μg/L for the trans-target analyte and 0.4, 2.3, and 2.8 μg/L for the cis-target analyte. The immunizing haptens, except for hapten 5, provided the target specific antibodies. Molecular modeling of the haptens supported the selection of reasonable immunizing haptens that best mimicked the target analyte. Hapten 5 was suitable as a coating antigen rather than as an immunogen since it had a different geometry. Very low cross-reactivities were measured to permethrin, its free metabolite (DCCA), PBA-glycine conjugate, and glycine. The ELISA will be optimized for the detection of total cis/trans-DCCA-glycine in human urine samples.

Original languageEnglish (US)
Pages (from-to)4583-4594
Number of pages12
JournalJournal of Agricultural and Food Chemistry
Volume52
Issue number15
DOIs
StatePublished - Jul 28 2004

Fingerprint

Permethrin
Pyrethrins
haptens
pyrethroid insecticides
Haptens
permethrin
antibody formation
Insecticides
Metabolites
immunoassays
Immunoassay
Antibody Formation
metabolites
Antibodies
glycine (amino acid)
Glycine
carboxylic acids
Carboxylic Acids
antigens
coatings

Keywords

  • ELISA
  • Glycine conjugate
  • Hapten
  • Human exposure
  • Permethrin

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Food Science
  • Chemistry (miscellaneous)

Cite this

Hapten and antibody production for a sensitive immunoassay determining a human urinary metabolite of the pyrethroid insecticide permethrin. / Ahn, Ki Chang; Watanabe, Takaho; Gee, Shirley J.; Hammock, Bruce D.

In: Journal of Agricultural and Food Chemistry, Vol. 52, No. 15, 28.07.2004, p. 4583-4594.

Research output: Contribution to journalArticle

@article{7958bfb891384133905d85134824e0d5,
title = "Hapten and antibody production for a sensitive immunoassay determining a human urinary metabolite of the pyrethroid insecticide permethrin",
abstract = "Permethrin is the most popular synthetic pyrethroid insecticide in agriculture and public health. For the development of the enzyme-linked immunosorbent assay (ELISA) to evaluate human exposure to permethrin, the glycine conjugate (DCCA-glycine) of a major metabolite, cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (DCCA), of permethrin was established as the target analyte. Four different types of the cis- and trans-isomers of immunizing haptens were synthesized as follows: N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine (hapten 3), N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1- carbonyl)-4-amino-L-phenyl-alanine (hapten 5), N-(N-(cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine)-amino-6-(2, 4-dinitrophenyl)aminohexanoic acid (hapten 9), and N-(cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine-4-oxobutanoic acid (hapten 24). Sixteen polyclonal antibodies produced against each cis- or trans-hapten-thyroglobulin conjugate as immunogens were screened against numerous hapten-bovine serum albumin conjugates as coating antigens. Six ELISAs with both a heterologous hapten structure and a heterologous hapten configuration (cis/trans or trans/cis) between antibody and coating antigen showed a high sensitivity for the target analyte. The IC50 was 1.3, 2.1, and 2.2 μg/L for the trans-target analyte and 0.4, 2.3, and 2.8 μg/L for the cis-target analyte. The immunizing haptens, except for hapten 5, provided the target specific antibodies. Molecular modeling of the haptens supported the selection of reasonable immunizing haptens that best mimicked the target analyte. Hapten 5 was suitable as a coating antigen rather than as an immunogen since it had a different geometry. Very low cross-reactivities were measured to permethrin, its free metabolite (DCCA), PBA-glycine conjugate, and glycine. The ELISA will be optimized for the detection of total cis/trans-DCCA-glycine in human urine samples.",
keywords = "ELISA, Glycine conjugate, Hapten, Human exposure, Permethrin",
author = "Ahn, {Ki Chang} and Takaho Watanabe and Gee, {Shirley J.} and Hammock, {Bruce D.}",
year = "2004",
month = "7",
day = "28",
doi = "10.1021/jf049646r",
language = "English (US)",
volume = "52",
pages = "4583--4594",
journal = "Journal of Agricultural and Food Chemistry",
issn = "0021-8561",
publisher = "American Chemical Society",
number = "15",

}

TY - JOUR

T1 - Hapten and antibody production for a sensitive immunoassay determining a human urinary metabolite of the pyrethroid insecticide permethrin

AU - Ahn, Ki Chang

AU - Watanabe, Takaho

AU - Gee, Shirley J.

AU - Hammock, Bruce D.

PY - 2004/7/28

Y1 - 2004/7/28

N2 - Permethrin is the most popular synthetic pyrethroid insecticide in agriculture and public health. For the development of the enzyme-linked immunosorbent assay (ELISA) to evaluate human exposure to permethrin, the glycine conjugate (DCCA-glycine) of a major metabolite, cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (DCCA), of permethrin was established as the target analyte. Four different types of the cis- and trans-isomers of immunizing haptens were synthesized as follows: N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine (hapten 3), N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1- carbonyl)-4-amino-L-phenyl-alanine (hapten 5), N-(N-(cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine)-amino-6-(2, 4-dinitrophenyl)aminohexanoic acid (hapten 9), and N-(cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine-4-oxobutanoic acid (hapten 24). Sixteen polyclonal antibodies produced against each cis- or trans-hapten-thyroglobulin conjugate as immunogens were screened against numerous hapten-bovine serum albumin conjugates as coating antigens. Six ELISAs with both a heterologous hapten structure and a heterologous hapten configuration (cis/trans or trans/cis) between antibody and coating antigen showed a high sensitivity for the target analyte. The IC50 was 1.3, 2.1, and 2.2 μg/L for the trans-target analyte and 0.4, 2.3, and 2.8 μg/L for the cis-target analyte. The immunizing haptens, except for hapten 5, provided the target specific antibodies. Molecular modeling of the haptens supported the selection of reasonable immunizing haptens that best mimicked the target analyte. Hapten 5 was suitable as a coating antigen rather than as an immunogen since it had a different geometry. Very low cross-reactivities were measured to permethrin, its free metabolite (DCCA), PBA-glycine conjugate, and glycine. The ELISA will be optimized for the detection of total cis/trans-DCCA-glycine in human urine samples.

AB - Permethrin is the most popular synthetic pyrethroid insecticide in agriculture and public health. For the development of the enzyme-linked immunosorbent assay (ELISA) to evaluate human exposure to permethrin, the glycine conjugate (DCCA-glycine) of a major metabolite, cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (DCCA), of permethrin was established as the target analyte. Four different types of the cis- and trans-isomers of immunizing haptens were synthesized as follows: N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine (hapten 3), N-(cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1- carbonyl)-4-amino-L-phenyl-alanine (hapten 5), N-(N-(cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine)-amino-6-(2, 4-dinitrophenyl)aminohexanoic acid (hapten 9), and N-(cis/trans-3-(2,2- dichlorovinyl)-2,2-dimethylcyclopropane-1-carbonyl)glycine-4-oxobutanoic acid (hapten 24). Sixteen polyclonal antibodies produced against each cis- or trans-hapten-thyroglobulin conjugate as immunogens were screened against numerous hapten-bovine serum albumin conjugates as coating antigens. Six ELISAs with both a heterologous hapten structure and a heterologous hapten configuration (cis/trans or trans/cis) between antibody and coating antigen showed a high sensitivity for the target analyte. The IC50 was 1.3, 2.1, and 2.2 μg/L for the trans-target analyte and 0.4, 2.3, and 2.8 μg/L for the cis-target analyte. The immunizing haptens, except for hapten 5, provided the target specific antibodies. Molecular modeling of the haptens supported the selection of reasonable immunizing haptens that best mimicked the target analyte. Hapten 5 was suitable as a coating antigen rather than as an immunogen since it had a different geometry. Very low cross-reactivities were measured to permethrin, its free metabolite (DCCA), PBA-glycine conjugate, and glycine. The ELISA will be optimized for the detection of total cis/trans-DCCA-glycine in human urine samples.

KW - ELISA

KW - Glycine conjugate

KW - Hapten

KW - Human exposure

KW - Permethrin

UR - http://www.scopus.com/inward/record.url?scp=3242766675&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3242766675&partnerID=8YFLogxK

U2 - 10.1021/jf049646r

DO - 10.1021/jf049646r

M3 - Article

C2 - 15264887

AN - SCOPUS:3242766675

VL - 52

SP - 4583

EP - 4594

JO - Journal of Agricultural and Food Chemistry

JF - Journal of Agricultural and Food Chemistry

SN - 0021-8561

IS - 15

ER -