Issue 2, 2007
From the journal:

Lab on a Chip

High-throughput screening of enzyme inhibition using an inhibitor gradient generated in a microchannel

Elena Garcia,a   Melissa S. Hasenbank,a   Bruce Finlaysonb  and  Paul Yager*a  

* Corresponding authors

a Department of Bioengineering, University of Washington, Box 355061, Foege N530J, 1705 NE Pacific Street, Seattle, WA 98195-2255, USA
E-mail: yagerp@u.washington.edu

b Department of Chemical Engineering, University of Washington, Box 355061, Foege N530J, 1705 NE Pacific Street, Seattle, WA 98195-2255, USA

Abstract

A new rapid microfluidic method for measuring enzyme inhibition is presented. The assay relies upon the creation of a uniform concentration of substrate and a microfluidically generated concentration gradient of inhibitor using a single microchannel and a single initial inhibitor concentration. The IC50 values of two enzyme inhibitors were determined using the new technique and validated using a conventional microtiter plate assay. Using both experimental and computational simulation techniques, the assay was shown to be sensitive to inhibitor potency and the distribution of inhibitor in the system. The method has the potential to be more accurate than conventional methods because of the comparatively large amount of data that may be collected. Recommendations for use of the assay are provided, including its use for high-throughput screening in drug discovery and general use in measurement of enzyme inhibition.

Graphical abstract: High-throughput screening of enzyme inhibition using an inhibitor gradient generated in a microchannel

About
Cited by
Related
Download options Please wait...

Article information

DOI
https://doi.org/10.1039/B608789B
Article type
Paper
Submitted
21 Jun 2006
Accepted
20 Sep 2006
First published
23 Oct 2006

Lab Chip, 2007,7, 249-255

Permissions

Request permissions

High-throughput screening of enzyme inhibition using an inhibitor gradient generated in a microchannel

E. Garcia, M. S. Hasenbank, B. Finlayson and P. Yager, Lab Chip, 2007, 7, 249 DOI: 10.1039/B608789B

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements