Fluorescence quenching competitive immunoassay in micro droplets

Jun Feng, Guomin Shan, Bruce D. Hammock, Ian M. Kennedy

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A fluorescence quenching competitive immunoassay in micro droplets was applied to the sensitive detection of the pyrethroid insecticide, esfenvalerate. Laser induced fluorescence from rhodamine dye was used as a marker. The competitive immunoreaction was performed in micro droplets generated by a vibrating orifice aerosol generator system with a 10-μm diameter orifice. Fluorescence that was emitted from the droplets was detected by a 1/8 m imaging spectrograph with a 512×512 thermoelectrically cooled, charged-coupled device camera. The conjugate of esfenvalerate with rhodamine exhibited similar fluorescence to that of pure rhodamine 6G. When anti-esfenvalerate antibodies were added to the droplets, the fluorescence decreased. The reduction in emission was due to a strong quenching effect that arises from the interaction between the protein and rhodamine molecules following the antigen-antibody reaction. When a sample of esfenvalerate was added to the droplets, the release of the conjugated rhodamine from the antigen-antibody complex allowed the fluorescence signal to recover. An assay in a picoliter droplet sample was shown to enable detection down to approximately 0.1 nM. A very small mass of analyte could be detected with this method. A sample of river water was used to gauge the impact of matrix effects and was shown to give rise to negligible interference with the immunoassay.

Original languageEnglish (US)
Pages (from-to)1055-1063
Number of pages9
JournalBiosensors and Bioelectronics
Volume18
Issue number8
DOIs
StatePublished - Aug 1 2003

Fingerprint

fenvalerate
Immunoassay
Quenching
Rhodamines
Fluorescence
Orifices
Antibodies
Antigen-antibody reactions
Antigen-Antibody Reactions
Pyrethrins
Insecticides
Gas generators
Spectrographs
Antigens
Antigen-Antibody Complex
Aerosols
Rivers
Gages
Anti-Idiotypic Antibodies
Assays

Keywords

  • Esfenvalerate
  • Fluorescence
  • Immunoassay
  • Micro droplet
  • Quenching

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Electrochemistry

Cite this

Fluorescence quenching competitive immunoassay in micro droplets. / Feng, Jun; Shan, Guomin; Hammock, Bruce D.; Kennedy, Ian M.

In: Biosensors and Bioelectronics, Vol. 18, No. 8, 01.08.2003, p. 1055-1063.

Research output: Contribution to journalArticle

Feng, Jun ; Shan, Guomin ; Hammock, Bruce D. ; Kennedy, Ian M. / Fluorescence quenching competitive immunoassay in micro droplets. In: Biosensors and Bioelectronics. 2003 ; Vol. 18, No. 8. pp. 1055-1063.
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