title: A flow-based enzyme-linked immunosorbent assay on a polydimethylsiloxane microchip for the rapid determination of immunoglobulin A.
abstract: A flow-based enzyme-linked immunosorbent assay (ELISA) on a polydimethylsiloxane (PDMS) microchip has been developed for the rapid determination of immunoglobulin A (IgA). The analytical principle of this integrated method is the same as the conventional sandwich-type ELISA. A primary antibody (anti-IgA) was adsorbed on the surface of a PDMS microchannel, and then an antigen (IgA) and a secondary antibody (anti-IgA HRP labeled) were reacted successively. The resulting antigen-antibody complex, fixed on the surface of the microchannel, was detected using Amplex((R)) Red and a fluorescent imaging system. The calibration curve of the IgA standard solution was linear in the range of 0-50ng/mL at the flow rate of 10muL/min. This flow rate corresponds to the reaction time of 4.8s. Compared to the conventional assay on a 96-well microtiter plate, the present assay on the microchip dramatically shortened the reaction time necessary for the enzyme-substrate reaction from 30min to 4.8s, i.e., to 1/375. The amounts of the reagent and sample were also reduced to 1/100 compared to the 96-well microtiter plate.
authors.affiliation: Faculty of Urban Environmental Sciences, Tokyo Metropolitan University, 1-1 Minamiohsawa, Hachioji, Tokyo 192-0397, Japan.
authors.forename: Hizuru
authors.initials: H
authors.lastname: Nakajima
authors.forename: Maiko
authors.initials: M
authors.lastname: Yagi
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kudo
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Nakagama
authors.forename: Takuya
authors.initials: T
authors.lastname: Shimosaka
authors.forename: Katsumi
authors.initials: K
authors.lastname: Uchiyama