期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 106, 期 1, 页码 335-342出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2004.08.025
关键词
mu-TAS; hydrogel; microfluidic device; enzyme reaction; fluorescence
A simple method to fabricate enzyme-containing microscopic hydrogel structures in microfluidic devices for the potential use in micro total analysis systems (mu=TAS) is described. Poly(ethylene glycol)-based hydrogel microstructures were prepared inside microchannels by photolithography and enzymes conjugated to a pH sensitive fluorophore (SNAFL-1) were incorporated into these hydrogel microstructures. Because of the ratiometric pH-dependent nature of SNAFL fluorescence, hydrogel microstructures exhibited a different emission intensity ratio with pH and this intensity ratio changed almost linearly between pH 7 and 12. When alkaline phosphatase-containing microreactors were exposed to p-nitrophenylphosphate (pNPP) as a substrate, phosphoric acid was produced inside the microstructure by enzymatic-catalyzed hydrolysis of the substrate and subsequently decreased the microenvironment pH. Because of the relatively rapid mass transport of analyte through the hydrogel, enzyme-catalyzed reaction was easily detected by change in emission intensity ratio before and after exposure to substrates. Enzyme-catalyzed reactions were quite fast and reached 90% of maximum value within 10 min. Data were analyzed using a modified Michae is-Menten equation and apparent Michaelis constants could be obtained. This system was also successfully applied to urea hydrolysis by urease. (c) 2004 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据