ISSN:
1040-0397
Keywords:
immunosensor
;
7-Hydroxycoumarin
;
Enzyme label
;
Amperometry
;
Chemistry
;
Polymer and Materials Science
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Chemistry and Pharmacology
Notes:
A regenerable immunosensor, based on a competitive assay principle and utilizing electrochemical detection, was developed for the determination of 7-hydroxycoumarin. Horseradish peroxidase (HRP) labeled anti-7-hydroxycoumarin antibody was employed in the determination, and the enzyme activity was measured as a function of current due to the reduction of hydrogen peroxide in the presence of a mediator (potassium ferrocyanide). Sensor preparation involved chemical immobilization of 7-hydroxy-coumarin-BSA conjugate on a glassy carbon electrode surface using a water soluble carbodiimide, 1 -ethyl-3(3-dimethyl-aminopropyl) carbodiimide, as coupling reagent in the presence of N-hydroxysuccinimide. The competition between the free and immobilized forms of the hapten for their HRP labeled antibody in solution allows for the quantitation of free 7-hydroxycoumarin. The sensor can be regenerated by simply immersing the electrode tip in acidic media. Under these conditions the hapten-antibody complex was selectively disrupted without adversely affecting the sensor performance. The conditions for removing the non-specific adsorption of the enzyme conjugate were also studied. The results showed that the detection range obtained for the hapten was physiologically relevant.
Additional Material:
4 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/elan.1140080703
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