Homogeneous substrate-labeled fluorescent immunoassay for carbamazepine

Area of Science:

• Clinical Chemistry
• Analytical Chemistry
• Biochemistry

Background:

• Carbamazepine is an anticonvulsant vital for epilepsy treatment.
• Its narrow therapeutic range necessitates precise serum monitoring to prevent toxicity.
• Existing monitoring methods can be complex or time-consuming.

Purpose of the Study:

• To develop a homogeneous substrate-labeled fluorescent immunoassay (SLFIA) for carbamazepine quantification in human serum.
• To establish a sensitive, reliable, and convenient method for therapeutic drug monitoring.
• To validate the SLFIA assay against established analytical techniques.

Main Methods:

• Synthesis of a carbamazepine fluorogenic reagent (FR).
• Utilizing beta-galactosidase for enzymatic hydrolysis producing fluorescence.
• Measuring fluorescence inhibition based on carbamazepine competition for antibody binding.

Main Results:

• The SLFIA assay demonstrated low within-run (1.6-5.5%) and between-run (1.9-3.5%) coefficients of variation.
• Assay results showed strong correlation with High-Performance Liquid Chromatography (HPLC) (r=0.97), Enzyme Immunoassay (r=0.98), and Gas Chromatography (GC) (r=0.95).
• The assay provides a sensitive and reproducible method for carbamazepine measurement.

Conclusions:

• The developed homogeneous SLFIA is a viable and accurate method for therapeutic drug monitoring of carbamazepine.
• This assay offers a convenient alternative for clinical laboratories managing epilepsy patients.
• SLFIA provides reliable carbamazepine serum concentrations, aiding in optimizing patient treatment and safety.