Redox cycling for an in-situ enzyme labeled immunoassay on interdigitated array electrodes.

摘要

This research is directed towards developing a more sensitive and rapid electrochemical sensor for enzyme labeled immunoassays by coupling redox cycling at interdigitated electrode arrays (IDA) with the enzyme label beta-galactosidase. Coplanar and comb IDA electrodes with a 2.4 mum gap were fabricated and their redox cycling currents were measured. ANSYS was used to model steady state currents for electrodes with different geometries. Comb IDA electrodes enhanced the signal about 3 times more than the coplanar IDAs, which agreed with the results of the simulation. Magnetic microbead-based enzyme assay, as a typical example of biochemical detection, was done using the comb and coplanar IDAs. The enzymes could be placed close to the sensing electrodes (∼10 mum for the comb IDAs) and detection took less than 1 min with a limit of detection of 70 amole of beta-galactosidase. We conclude that faster and more sensitive assays can be achieved with the comb IDA. A paramagnetic bead assay has also been demonstrated for detection of bacteriophage MS2, used as a simulant for biothreat viruses, such as small pox. The immunoassay was carried out in a microfluidic format with the IDA, reference and counter electrodes integrated on the same chip. Detection of 90 ng/mL MS2 or 1.5 x 1010 MS2 particles/mL was demonstrated.