Development of a noncompetitive phage anti-immunocomplex assay for brominated diphenyl ether 47

Hee Joo Kim, Martin A. Rossotti, Ki Chang Ahn, Gualberto G. González-Sapienza, Shirley J. Gee, Ruthie Musker, Bruce D. Hammock

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

We present a new application of the noncompetitive phage anti-immunocomplex assay (PHAIA) by converting an existing competitive assay to a versatile noncompetitive sandwich-type format using immunocomplex binding phage-borne peptides to detect the brominated flame retardant, brominated diphenyl ether 47 (BDE 47). Three phage-displayed 9-mer disulfide-constrained peptides that recognize the BDE 47-polyclonal antibody immunocomplex were isolated. The resulting PHAIAs showed variable sensitivities, and the most sensitive peptide had a dose-response curve with an SC50 (concentration of analyte producing 50% saturation of the signal) of 0.7ng/ml BDE 47 and a linear range of 0.3-2ng/ml, which was nearly identical to the best heterologous competitive format (IC50 of 1.8ng/ml, linear range of 0.4-8.5/ml). However, the PHAIA was 1400-fold better than homologous competitive assay. The validation of the PHAIA with extracts of house furniture foam as well as human and calf sera spiked with BDE 47 showed overall recovery of 80-113%. The PHAIA was adapted to a dipstick format (limit of detection of 3.0ng/ml), and a blind test with six random extracts of local house furniture foams showed that the results of the PHAIA and dipstick assay were consistent, giving the same positive and negative detection.

Original languageEnglish (US)
Pages (from-to)38-46
Number of pages9
JournalAnalytical Biochemistry
Volume401
Issue number1
DOIs
StatePublished - Jun 2010

Keywords

  • BDE 47
  • Brominated flame retardant
  • Noncompetitive immunoassay
  • Phage anti-immunocomplex assay
  • Phage ELISA
  • Phage-displayed peptide

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Kim, H. J., Rossotti, M. A., Ahn, K. C., González-Sapienza, G. G., Gee, S. J., Musker, R., & Hammock, B. D. (2010). Development of a noncompetitive phage anti-immunocomplex assay for brominated diphenyl ether 47. Analytical Biochemistry, 401(1), 38-46. https://doi.org/10.1016/j.ab.2010.01.040