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An on-line immunoassay method for theophylline using a protein A immunoreactor
P Fernandez-Hernando1, J N Miller
1Department of Chemistry, Loughborough University of Technology, Leicestershire, UK.
Journal of Pharmaceutical and Biomedical Analysis
|January 1, 1991
Summary
This study automated a fluorescence immunoassay for theophylline using flow injection analysis. The system efficiently separates antibody-bound theophylline, enabling precise quantification at microgram levels.
Area of Science:
- Analytical Chemistry
- Biochemistry
Background:
- Theophylline measurement is crucial for therapeutic drug monitoring.
- Automated immunoassays improve efficiency and reduce errors in clinical diagnostics.
Purpose of the Study:
- To automate a heterogeneous fluorescence immunoassay for theophylline determination.
- To develop a robust flow injection analysis (FIA) system for rapid theophylline quantification.
Main Methods:
- Utilized a flow injection analysis system with a protein A solid-phase reactor.
- Employed a heterogeneous fluorescence immunoassay principle for antibody-antigen separation.
- Optimized reaction conditions including pH, flow rate, and reactant concentrations.
Main Results:
- Successfully automated theophylline immunoassay using FIA.
- Achieved sensitive theophylline detection at the micrograms per milliliter level.
- Demonstrated repeated cycling of the protein A reactor between neutral and acid pH for continuous analysis.
Conclusions:
- The automated FIA system provides a sensitive and efficient method for theophylline analysis.
- The protein A solid-phase reactor is effective for separating antibody-bound theophylline in a continuous flow system.
- This automation strategy holds promise for routine clinical laboratory applications.