Sensitive electrochemical enzyme immunoassay microdevice based on architecture of dual ring electrodes with a sensing cavity chamber

A novel electrochemical detection architecture was investigated for enzyme immunoassay sensors. Microchips with dual-ring working and counter electrodes, and a sensing cavity chamber were made on glass slides. The glass surface of the microchip was coated by 3-aminopropyltriethoxysilane (APTES). Goat IgG, as a example, was covalently captured on APTES-modified glass surfaces through glutaraldehyde (GA) as a cross-linker. Enzyme substrate, p-aminophenyl phosphate (PAPP) was prepared by electrolysis. The enzyme conversion from home-synthetic PAPP to p-aminophenol (PAP) was examined by differential pulse voltammetry (DPV). A competitive inhibition enzyme-linked immunosorbant assay (ELISA) was designed to test the system. Experimental results demonstrate that a detection limit of 118 fg/ml of goat IgG and a dynamic range of 118 fg/ml to 1.18 ng/ml, up to five orders of magnitude could be achieved. Due to its novel architecture design and electronic detection scheme, the method can be used to fabricate portable electrochemical ELISA lab-on-chip systems. The technology could have great potential in clinical diagnostic applications.