Enzyme linked immunosorbent assay on a microchip with electrochemical detection

Abstract

This paper presents the development of a sandwich immunoassay in disposable plastic microchips. Photoablated microchannels with integrated electrodes have been used for the development of enzyme-linked-immunosorbent-assay (ELISA). The presence of the electrode inside the 40 nL microchannel enables the detection of the redox active enzyme substrate directly inside the reaction channel. Furthermore, due to the small diffusion distances, each incubation time can be reduced to five minutes instead of a few hours in standard microtiterplates. The initial characterisation of this immunoassay has been performed with a large protein complex D-Dimer-alkaline phosphatase. This system was used for the detection of immobilised antibodies on the surface of the photoablated microchannel. In a second step, a sandwich immunoassay with a horseradish peroxidase-secondary antibody conjugate (HRP-conjugate) was used to detect D-Dimer between 0.1 and 100 nM, which is the relevant concentration range of the clinical tests.