Magnetic Bead-Based Phage Anti-Immunocomplex Assay (PHAIA) for the Detection of the Urinary Biomarker 3-Phenoxybenzoic Acid to Assess Human Exposure to Pyrethroid Insecticides

Details

• Personal Author:
  Ahn KC ; Gee SJ ; González-Sapienza GG ; González-Techera A ; Hammock BD ; Kim H-J
• Description:
  Noncompetitive immunoassays are advantageous over competitive assays for the detection of small molecular weight compounds. We recently demonstrated that phage peptide libraries can be an excellent source of immunoreagents that facilitate the development of sandwich-type noncompetitive immunoassays for the detection of small analytes, avoiding the technical challenges of producing anti-immunocomplex antibody. In this work we explore a new format that may help to optimize the performance of the phage anti-immunocomplex assay (PHAIA) technology. As a model system we used a polyclonal antibody to 3-phenoxybenzoic acid (3-PBA) and an anti-immunocomplex phage clone bearing the cyclic peptide CFNGKDWLYC. The assay setup with the biotinylated antibody immobilized onto streptavidin-coated magnetic beads significantly reduced the amount of coating antibody giving identical sensitivity (50% saturation of the signal (SC(50))=0.2-0.4ng/ml) to the best result obtained with direct coating of the antibody on ELISA plates. The bead-based assay tolerated up to 10 and 5% of methanol and urine matrix, respectively. This assay system accurately determined the level of spiked 3-PBA in different urine samples prepared by direct dilution or clean-up with solid-phase extraction after acidic hydrolysis with overall recovery of 80-120%. [Description provided by NIOSH]
• Subjects:
  Antibodies Biochemistry Biomarkers Chemistry Enzymes Immune System Occupational Health Safety
• Keywords:
  3-Phenoxybenzoic Acid Antibody Response Author Keywords: Phage Anti-immunocomplex Assay Chemical Analysis ELISA Enzyme-linked Immunosorbent Assay Immunology Noncompetitive Immunoassay Phage ELISA Phage Peptide Display Pyrethroid Insecticides Quantitative Analysis

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• ISSN:
  0003-2697
• 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
• Pages in Document:
  45-52
• Volume:
  386
• Issue:
  1
• NIOSHTIC Number:
  nn:20034969
• Citation:
  Anal Biochem 2009 Mar; 386(1):45-52
• Contact Point Address:
  Bruce D. Hammock, Department of Entomology and UCD Cancer Research Center, University of California, Davis, CA 95616
• Email:
  bdhammock@ucdavis.edu
• Federal Fiscal Year:
  2009
• NORA Priority Area:
  Agriculture, Forestry and Fishing
• Performing Organization:
  University of California - Davis
• Peer Reviewed:
  True
• Start Date:
  20010930
• Source Full Name:
  Analytical Biochemistry
• End Date:
  20270929
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:1a5a92fe104e64e4c239cb13c9a6ce2f7345b6ed33a0704b469b445422da1ffb1c095e749937fa9c31eac7b479ca89a75a7e8f0dcf3f6e1ec712a5d43ea7542f
• Download URL:
  https://stacks.cdc.gov/view/cdc/186976/cdc_186976_DS1.pdf
• File Type:
  [PDF - 476.09 KB ]