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High-throughput Enzyme Assay on a Multichannel Microchip Using Optically Gated Sample Introduction

Overview
Journal Electrophoresis
Specialty Chemistry
Date 2005 Nov 19
PMID 16294297
Citations 4
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Abstract

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.

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References
1.
Auroux P, Iossifidis D, Reyes D, Manz A . Micro total analysis systems. 2. Analytical standard operations and applications. Anal Chem. 2002; 74(12):2637-52. DOI: 10.1021/ac020239t. View

2.
Huang Z . Kinetic assay of fluorescein mono-beta-D-galactoside hydrolysis by beta-galactosidase: a front-face measurement for strongly absorbing fluorogenic substrates. Biochemistry. 1991; 30(35):8530-4. DOI: 10.1021/bi00099a005. View

3.
Burns M, Mastrangelo C, Sammarco T, Man F, Webster J, Johnsons B . Microfabricated structures for integrated DNA analysis. Proc Natl Acad Sci U S A. 1996; 93(11):5556-61. PMC: 39285. DOI: 10.1073/pnas.93.11.5556. View

4.
Medintz I, Paegel B, Blazej R, Emrich C, Berti L, Scherer J . High-performance genetic analysis using microfabricated capillary array electrophoresis microplates. Electrophoresis. 2001; 22(18):3845-56. DOI: 10.1002/1522-2683(200110)22:18<3845::AID-ELPS3845>3.0.CO;2-0. View

5.
Shi Y, Simpson P, Scherer J, Wexler D, Skibola C, Smith M . Radial capillary array electrophoresis microplate and scanner for high-performance nucleic acid analysis. Anal Chem. 1999; 71(23):5354-61. DOI: 10.1021/ac990518p. View