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Automated amplified flow immunoassay for cocaine
C G Bauer1, A V Eremenko, A Kühn
1Institute of Biochemistry and Molecular Physiology, University of Potsdam, Luckenwalde, Germany.
Analytical Chemistry
|November 21, 1998
Summary
A novel amplified flow immunoassay (AFIA) enables rapid, picomolar detection of cocaine. This method combines immunoenzymometric assay with a biosensor for sensitive and efficient analysis of cocaine concentrations.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Biomedical Engineering
Background:
- Hapten detection requires sensitive and rapid analytical methods.
- Traditional immunoassays can lack the sensitivity or speed for certain applications.
- Enzyme labels offer potential for signal amplification in immunoassays.
Purpose of the Study:
- To develop and validate an amplified flow immunoassay (AFIA) for the sensitive detection of cocaine.
- To integrate a noncompetitive immunoenzymometric assay (IEMA) with an on-line biosensor for real-time enzyme label detection.
- To establish the analytical performance characteristics of the AFIA for cocaine analysis.
Main Methods:
- Development of an AFIA combining IEMA with an alkaline phosphatase (ALP) label.
- Utilized an affinity column with immobilized cocaine for separation of unbound antibody-label complexes.
- Employed an amperometric substrate-recycling biosensor with tyrosinase and glucose dehydrogenase for ALP detection.
Main Results:
- Achieved a lower limit of detection of 380 pM (38 fmol) for cocaine.
- Demonstrated a sampling rate of 26 samples per hour.
- Showed imprecision of +/- 10% and recovery of 49 +/- 3% in real samples.
Conclusions:
- AFIA is a viable approach for the rapid detection of picomolar concentrations of haptens like cocaine.
- The integrated biosensor system provides sensitive and efficient on-line detection of enzyme labels.
- This method offers a significant advancement in analytical techniques for small molecule detection.