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Permanently oriented antibody immobilization for digoxin determination with a flow-through fluoroimmunosensor
P Fernández1, J S Durand, C Pérez-Conde
1Dpto. Ciencias Analíticas, Facultad de Ciencias, Universidad Nacional de Educación a Distancia (UNED), C/ Senda del Rey n masculine 9, 28040, Madrid, Spain. pfhernando@ccia.uned.es
Analytical and Bioanalytical Chemistry
|May 7, 2003
Summary
A novel fluoroimmunosensor utilizing antibodies on controlled-pore glass (CPG) accurately detects digoxin, a heart disease medication. This method offers high reproducibility and minimal cross-reactivity, proving effective for human serum analysis.
Area of Science:
- Analytical Chemistry
- Biomedical Engineering
- Immunosensor Technology
Background:
- Digoxin is crucial for treating congestive heart failure and artery disease.
- Accurate and sensitive detection of digoxin is vital for patient management.
- Existing detection methods may have limitations in speed, sensitivity, or specificity.
Purpose of the Study:
- To develop and validate a new flow-through fluoroimmunosensor for digoxin determination.
- To assess the performance characteristics of the developed immunosensor, including detection limit, reproducibility, and working range.
- To evaluate the specificity and practical applicability of the fluoroimmunosensor in human serum samples.
Main Methods:
- Immobilization of antibodies onto a controlled-pore glass (CPG) reactor.
- Development of a flow-through fluoroimmunoassay system.
- Testing for detection limit, reproducibility, working concentration range, and cross-reactivity with structurally similar compounds.
- Validation using human serum samples.
Main Results:
- The fluoroimmunosensor achieved a detection limit of 1.20 microg L(-1).
- High reproducibility was observed (RSD=4.5% at 0.0025 mg L(-1), 6.7% at 0.01 mg L(-1)).
- The optimal working range was 1.2 x 10(-3)-4.0 x 10(-2) mg L(-1) with no cross-reactivity at a 1:100 ratio with similar structures.
- The immunosensor demonstrated a lifetime of approximately 50 assays or three months when stored, with a sample throughput of 10-12 samples/hour.
Conclusions:
- The developed flow-through fluoroimmunosensor provides a sensitive, reproducible, and specific method for digoxin determination.
- The immunosensor is suitable for practical application in analyzing digoxin levels in human serum.
- This technology offers a potential advancement in therapeutic drug monitoring for digoxin therapy.