부산대학교 도서관

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.talanta.2005.12.059 Byline: Hizuru Nakajima (a), Maiko Yagi (a), Yuki Kudo (a), Tatsuro Nakagama (a), Takuya Shimosaka (b), Katsumi Uchiyama (a) Keywords: Microchip; ELISA; Rapid determination; Immunoglobulin A; Saliva; Stress 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.sup.[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 10[mu]L/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. Author Affiliation: (a) Faculty of Urban Environmental Sciences, Tokyo Metropolitan University, 1-1 Minamiohsawa, Hachioji, Tokyo 192-0397, Japan (b) National Metrology Institute of Japan (NMIJ), National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 3, Umezono, Tsukuba, Ibaraki 305-8563, Japan Article History: Received 28 October 2005; Revised 17 December 2005; Accepted 22 December 2005