Journal Article


R O'Kennedy
M R Smyth
E Dempsey
B Deasy
D A Egan



enzyme linked immunosorbent assay coumarins chromatography high pressure liquid electrochemistry rabbits electrophoresis polyacrylamide gel diagnostic use male 7 hydroxycoumarin serum albumin bovine coumarin umbelliferones thyroglobulin ovalbumin isolation purification binding competitive humans metabolism antigen antibody reactions antibodies animals immunology antibody specificity urine

Antibody-based approaches to coumarin analysis. (1994)

Abstract 7-Hydroxycoumarin (7-HC) was chemically conjugated by diazo coupling to carrier proteins such as bovine serum albumin (BSA), thyroglobulin and ovalbumin. These conjugates were characterised by sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS-PAGE) and high-performance liquid chromatography (HPLC). Rabbits were immunised using the 7-HC-BSA conjugate. The highest antibody titre achieved was 1:10,000, as determined by competitive enzyme-linked immunosorbent assay (ELISA). The resulting antibodies were purified by ammonium sulphate precipitation, followed by protein A affinity chromatography. Their purity was assessed by SDS-PAGE and HPLC. These antibodies have been used in the development of a competitive ELISA, an amperometric biosensor and an electrochemical immunoassay. Both the ELISA and amperometric biosensor have been successfully applied to the analysis of 7-HC and its glucuronide conjugate in human urine samples. Each of these antibody-based methods provides a novel approach to the analysis of the main metabolites of coumarin.
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Full list of authors on original publication

R O'Kennedy, M R Smyth, E Dempsey, B Deasy, D A Egan

Experts in our system

R O'Kennedy
Dublin City University
Total Publications: 211
Eugene M Dempsey
University College Cork
Total Publications: 132