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Flow injection electrochemical enzyme immunoassay for theophylline using a protein A immunoreactor and p-aminophenyl
D A Palmer1, T E Edmonds, N J Seare
1Department of Chemistry, Loughborough University of Technology, Leicestershire, UK.
Abstract:
A competitive electrochemical enzyme immunoassay has been developed for the antiasthmatic drug theophylline, utilizing a controlled-pore glass-protein A immunoreactor and flow injection techniques. p-Aminophenyl phosphate, a substrate for alkaline phosphatase, has been used in this assay, and its hydrolysis product p-aminophenol was determined at +0.2 V versus the saturated calomel electrode. For each sample the antibody-protein A reaction takes place at near-neutral pH, and the complexes are eluted at acid pH. Serum theophylline has been determined by this method, and good relative standard deviations and percentage recoveries have been achieved.