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Effect of Antibodies Immobilization on the Efficiency of Carbon Paste Based Immunosensors

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Abstract:

Carbon paste electrodes are widely used in realizing immunosensors. In the present work, two types of immobilization procedures were employed. In one method the antibodies were immobilized on the surface of carbon paste electrode (SAE). In another method the antibodies were incorporated in to the carbon paste itself. Thus immunocomposite (ICE) was made and it was used as renewable immunosensor. Both of these electrodes showed similar behavior in cyclic voltammetric experiments. In order to compare these two methods of immobilization, a model system comprising rabbit anti-mouse immunoglobulin G (RαMIgG) as the capturing antibody, mouse IgG (MIgG) as antigen and alkaline phosphatase conjugated to RαMIgG as revealing antibody was used. The measurement procedure involved sandwich enzyme linked immunosorbent assay (ELISA) principle. 1-naphthyl phosphate was used as the enzymatic substrate. 1-naphthol, the enzymatic hydrolysis product of substrate, was measured amperometrically at a potential of +400 mV versus Ag/AgCl reference electrode. The above methods of immobilization were compared for their sensitivity, ease of preparation of electrode, amount of capturing antibody required, total assay time and shelf life. The SAE has high sensitivity and superior to ICE.

Keywords: AMPEROMETRY; CARBON PASTE ELECTRODE; ELISA; IMMUNOCOMPOSITE; IMMUNOSENSING; MOUSE IGG; PHYSICAL ADSORPTION

Document Type: Research Article

DOI: http://dx.doi.org/10.1166/sl.2009.1118

Publication date: August 1, 2009

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