Hapten and Antibody Production for a Sensitive Immunoassay Determining a Human Urinary Metabolite of the Pyrethroid Insecticide Permethrin

Details

• Personal Author:
  Ahn KC ; Gee SJ ; Hammock BD ; Watanabe T
• Description:
  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-phenylalanine (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 microg/L for the trans-target analyte and 0.4, 2.3, and 2.8 microg/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. [Description provided by NIOSH]
• Subjects:
  Agriculture Chemistry, Analytic Enzymes Insecticides Occupational Health Safety
• Keywords:
  Agricultural Industry Analytical Chemistry Analytical Methods Author Keywords: ELISA ELISA Enzyme-linked Immunosorbent Assay Glycine Conjugate Hapten Human Exposure Permethrin Pesticides And Agricultural Chemicals

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• ISSN:
  0021-8561
• Document Type:
  Text
• Funding:
  Cooperative Agreement
• Genre:
  Journal Article
• Place as Subject:
  California ; OSHA Region 9
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  52
• Issue:
  15
• NIOSHTIC Number:
  nn:20028901
• Citation:
  J Agric Food Chem 2004 Jul; 52(15):4583-4594
• Contact Point Address:
  Department of Entomology and UCD Cancer Research Center, University of California, Davis, CA 95616, USA
• CAS Registry Number:
  Permethrin (CAS RN 52645-53-1)
• Federal Fiscal Year:
  2004
• Performing Organization:
  University of California - Davis
• Peer Reviewed:
  True
• Start Date:
  20010930
• Source Full Name:
  Journal of Agricultural and Food Chemistry
• End Date:
  20270929
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:8e9d80c6efd9c937c55ec117ffd20bc0488e07380a9e3280582cefc901300675082ce0881269c9aa579b9ad77614dcac7d45a85d5373c96e5f285910f98d8c1f
• Download URL:
  https://stacks.cdc.gov/view/cdc/189994/cdc_189994_DS1.pdf
• File Type:
  [PDF - 230.54 KB ]