earticle

영어

In this work, a novel microfluidic device was prepared for the enzyme-based bioassay. Microfluidic device had two serial chambers connected by microchannels, reaction chamber and detection chamber. The reaction chamber was filled with glass beads with immobilized enzymes via APTES. Key feature of first chamber was filters on both sides, which was used to retain the beads with D of 40-70 µm in the flowing system. In the second chamber, PEG hydrogel microarray was fabricated via photolithography, which could immobilize proteins or fluorescent dyes for the optical analysis of the reaction. As a model, various concentration of glucose was detected in the system. In this system, first chamber was filled with GOX-immobilized beads, and second chamber with HRP-entrapped hydrogel microarray. When solution of glucose and Amplex Red flowed into channels, glucose reacted with GOX on the beads to produce H2O2, which moved to the second chamber and diffused into hydrogel array with HRP. Next, Amplex Red reacts with H2O2 in the presence of HRP inside array to produce fluorescent resorufin. By measuring the fluorescence of resorufin, glucose concentration ranging from 0.1–50 mM could be detected.