Enhanced Luminescent Quantitation of Horseradish Peroxidase Conjugates: Application in an Enzyme Immunoassay for Digoxin

Abstract

A p-iodophenol-enhanced luminescent end-point has been incorporated into a commercially available heterogeneous competitive enzyme immunoassay for digoxin based on a horseradish peroxidase-digoxin conjugate. The luminescent end-point could be completed in less than 1 min and significantly reduced overall assay time. Results for the assay obtained using enhanced luminescence showed good agreement with those obtained using a colorimetric end-point (correlation coefficient 0·98). Both assays gave acceptable precision within the therapeutic range. The incubation time for the luminescent immunoassay was reduced to 15 min and still gave differentiation between sub-therapeutic, therapeutic and toxic levels of digoxin.