Adsorption of immunoglobulin G on carbon paste electrodes as a basis for the development of immunoelectrochemical devices.
The attachment of Immunoglobulin G (IgG) to a carbon paste electrode is investigated in this work. Studies of the immobilization of the immunoglobulin on this electrode were carried out. Alkaline phosphatase (AP) has been used as an enzyme label, linked to the antibody, for the determination of the adsorbed immunoglobulin. Various substrates for this enzyme have been tested and alpha-naphthyl phosphate was found to be the best because of the lower oxidation potential of the enzymatic product, alpha-naphthol, and greater sensitivity under the same experimental conditions. An electrodic pretreatment based on an anodic oxidation of the electrode surface in a phosphate media pH 9 was carried out prior to the adsorption step. This activation is also suitable for the removal of the IgG layer and the regeneration of the electrodic surface after each determination, with the intention of using the same electrode in the subsequent assays. A good reproducibility of the signal was achieved in this way (Relative Standard Deviation, R.S.D. = 3.7%). Using cyclic voltammetry, IgG labelled with AP has been quantified in a range from 2 x 10(-12) to 7 x 10(-11) M and a detection limit of 2.3 x 10(-12) M (signal-to-noise ratio = 3) was found. Finally, human IgG was quantified under non-optimized conditions on the electrodic surface through the reaction with AP labelled IgG using two different immunological designs.
Fernández-Sánchez
C
C
Departamento de Química Física y Analítica, Universidad de Oviedo, Asturias, Spain.
Costa-García
A
A
D000469:Alkaline Phosphatase / Q000032:analysis; D015374:Biosensing Techniques; D004563:Electrochemistry / Q000295:instrumentation*; D004566:Electrodes; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D007074:Immunoglobulin G / Q000032:analysis*
D007074:Immunoglobulin G; D000469:Alkaline Phosphatase