Development of an antibody-based amperometric biosensor to study the reaction of 7-hydroxycoumarin with its specific antibody.
An amperometric biosensor, based on an anti-7-hydroxycoumarin (7-OH-coumarin) antibody immobilized at the surface of a glassy carbon electrode, and contained behind a cellulose dialysis membrane, is described. The electrochemical behaviour of this metabolite at the bare glassy carbon electrode was found to be well defined by using differential-pulse voltammetry, and a 90% decrease in peak height was observed on binding of the antibody to the antigen, which occurs at the electroactive site of 7-OH-coumarin. The kinetics of the antibody-antigen reaction was investigated and a first-order reaction was observed with k = 0.0329 mA min-1. This system provides a novel method for studying antibody specificity and the kinetics of such antibody-antigen interactions.
School of Chemical Sciences, Dublin City University, Ireland.
E
E
Dempsey
C
C
O'Sullivan
M R
MR
Smyth
D
D
Egan
R
R
O'Kennedy
J
J
Wang
D000906:Antibodies; D000918:Antibody Specificity; D015374:Biosensing Techniques; D004563:Electrochemistry; D004566:Electrodes; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D014468:Umbelliferones / Q000737:chemistry / Q000276:immunology*
D000906:Antibodies; D014468:Umbelliferones; C031477:7-hydroxycoumarin