Sequential injection (SI) system incorporated with amperometric immunosensor was developed for sensitive determination of human immunoglobulin G (HIgG). A cost effective label-free immunosensor was fabricated by immobilizing anti-HIgG on a graphene oxide (GO) modified screen-printed carbon electrode (SPCE). The developed electrode was characterized by cyclic voltammetry(CV), scanning electron microscope(SEM), and energy dispersive spectroscopy(EDS) which confirmed the selective immunointeraction of HIgG to the anti-HIgG on the electrode, thus reduced the amperometric current of [Fe(CN)] redox probe. The sensing electrode was placed in a designed electrochemical flow cell of SI system, where the redox probe was propelled through and the currents before and after the immunointeraction occurred were measured amperometrically by using a simple home-made amperometer. Under the optimum condition: flow rate of 2mLmin, applied potential of +350mV, [Fe(CN)] concentration of 10mM and 10min of incubation time, a linear calibration in the range of 2-100ngmL was achieved, with detection limit of 1.70ngmL. The proposed system provided good repeatability and reproducibility and the application for urine sample analysis was demonstrated.
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