ELISA for the quantification of pilsicainide

Area of Science:

• Analytical Chemistry
• Pharmacology

Background:

• Pilsicainide is an antiarrhythmic drug requiring precise quantification.
• Existing analytical methods may lack the sensitivity or practicality for certain applications.

Purpose of the Study:

• To develop and validate a sensitive and specific enzyme-linked immunosorbent assay (ELISA) for quantifying pilsicainide.
• To assess the assay's performance against high-performance liquid chromatography (HPLC).

Main Methods:

• Development of an anti-pilsicainide antibody through rabbit immunization with a pilsicainide-bovine serum albumin conjugate.
• Enzyme labeling of pilsicainide using beta-D-galactosidase.
• Assay validation including specificity, sensitivity, and cross-reactivity assessments.
• Comparison of ELISA results with HPLC measurements.

Main Results:

• The developed ELISA demonstrated high sensitivity, measuring pilsicainide concentrations as low as 1.6 ng/ml.
• The antibody exhibited specificity for pilsicainide, recognizing key structural moieties.
• ELISA results showed good correlation with HPLC measurements.
• The ELISA was approximately 30-fold more sensitive than HPLC for detecting low pilsicainide concentrations.
• Drug levels were successfully measured in rabbit serum after oral administration.

Conclusions:

• The developed ELISA is a sensitive and specific method for pilsicainide quantification.
• This assay is a valuable tool for therapeutic drug monitoring (TDM) and pharmacokinetic studies of pilsicainide.
• The ELISA offers improved sensitivity for detecting low drug concentrations.