High-throughput enzyme assay on a multichannel microchip using optically gated sample introduction.
To meet the requirements for high-throughput screening for drug discovery research, it is very important to develop techniques with the ability of performing multiple enzyme assays simultaneously. Using optically gated sample introduction on a multichannel microchip, multiple enzyme assays have been demonstrated in four parallel channels. The hydrolysis of fluorescein mono-beta-D-galactopyranoside by beta-galactosidase and the inhibition of this reaction by the competitive inhibitor phenylethyl beta-D-thiogalactoside were initially studied to determine the effect of system movement using the voice coil actuator on the enzyme assay reaction. The results from these two studies are consistent with the results from the assay using a single-channel microchip, and they demonstrate that the system using optically gated sample introduction on multichannel microchip can be used to perform multiple enzyme assays. Three unique enzyme assays were also performed in different channels, which show this technique could be competitive for high-throughput screening in drug discovery with other traditional techniques.
Xu
Hongwei
H
152 Davey Laboratory, Department of Chemistry, Pennsylvania State University, University Park, PA 16802, USA.
Ewing
Andrew G
AG
D000818:Animals; D002417:Cattle; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005452:Fluoresceins / Q000737:chemistry; D005697:Galactosides / Q000737:chemistry; D007700:Kinetics; D007785:Lactose / Q000494:pharmacology; D008099:Liver / Q000201:enzymology; D055095:Optics and Photonics / Q000295:instrumentation*; D013862:Thiogalactosides / Q000494:pharmacology; D001616:beta-Galactosidase / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism
D004791:Enzyme Inhibitors; D005452:Fluoresceins; D005697:Galactosides; D013862:Thiogalactosides; C070825:fluorescein monogalactoside; C066907:phenylethylthio-beta-galactopyranoside; D001616:beta-Galactosidase; D007785:Lactose