Caliper Life Sciences, Inc.
68 Elm Street
Hopkinton, MA, 01748
Website: http://www.caliperLS.com
|
 |
Microfluidic Kinetic Assays for Protein and Lipid Kinase and Phosphatase Targets
by Abbie Esterman PhD and Seth P. Cohen PhD
Introduction
Much of the effort to find new drugs has focused on protein kinases and phosphatases. Many of these targets play important roles in the modulation of cell metabolism and disease. The elucidation of the role of lipids in cellular metabolism, obesity, inflammation, heart disease, and cancer has increased the interest of pharmaceutical, biotechnology and academic scientists to include lipid targets in their quest for new drugs. Scientists working in drug discovery are faced with the task of developing these new assays which must be robust and meet a list of demanding criteria. To achieve these criteria, the assays must be rapid, economical, miniaturized, automated, sensitive and precise. Traditional assay formats, particularly those used for enzymatic screening, can be quite complex and many require specialized reagents such as radiolabeled substrates, specific antibodies, or other detection components. A number of the existing technologies for lipid assays use readouts such as HPLC which are not amenable to the higher throughput needed for compound screening. Furthermore, the characterization of enzyme activity through accurate determination of substrate Km and inhibitor Ki requires the ability to measure the rates of product formation, and many of the methodologies used for identifying active compounds that modulate enzymatic activity are limited to endpoint assays and cannot easily be used to follow a reaction over time.
Due to the equations in this article, we have made a PDF
available for you to download.
|
|
