LabAutomation 2006 - SLAS

MP09
Alex Batchelor
Cambrex Bio Science Nottingham
Nottingham, United Kingdom
alex.batchelor@cambrex.com
Where Laboratory Technologies Emerge and Merge
Co-Author(s)
Lee Walker
Anthony Pitt
PDELight - A Novel, Generic and Simple High Throughput Assay for
Screening cAMP-Dependent Phosphodiesterases
In the cell phosphodiesterases (PDEs) function in conjunction with adenylate cyclases to regulate the amplitude of the ubiquitous
2nd messenger signalling molecule, cyclic adenosine monophosphate (cAMP). PDEs catalyze the hydrolysis of cAMP to adenosine
monophosphate (AMP). There are at least 11 different families of PDEs most of which contain more than one isozyme. Their substrate
specificities, kinetics and tissue specific expression make PDEs drugable targets for a range of diseases.
A number of HTS assays are used to identify inhibitors of cAMP-PDEs. However these are either rad-based, require the use of beads,
modified substrate or antibodies and are time consuming to perform.
We introduce a novel luminescent HTS assay which offers a simple alternative to the current cAMP-PDE assays. The AMP produced
from PDE hydrolysis of cAMP is quantified using a robust and highly sensitive luciferase-based luminescent reagent. The AMP is directly
converted to ATP and quantified as light. Nearly a photon of light is emitted for every molecule of AMP produced. The assay is extremely
simple to use and can be run in a number of ways to suit the user.
Data represented in this study demonstrates the principle and performance of the assay.
MP10
Michael Benedetti
Buck Institute
Novato, California
mbenedetti@buckinstitute.org
Co-Author(s)
Matthew Gill
Anders Olsen
Amanda Foster
Gordon Lithgow
Development of High-Throughput Screens for Anti-Aging Compounds in the Nematode
Caenorhabditis Elegans
The nematode Caenorhabditis elegans provides an excellent model organism for investigating the aging process. These worms have a
short lifespan of approximately 20 days and many single gene mutations have been found that more than double their lifespan. There
is now considerable interest in identifying drugs that can influence nematode lifespan as they may provide novel therapeutic targets
for amelioration of age related disease. Small scale, targeted screens have demonstrated that nematode lifespan can be increased by
treatment with drugs such as anti-oxidants and anticonvulsants. It is likely that the development of high throughput screening methods will
facilitate the discovery of many more compounds that are able to extend the lifespan of C. elegans.
C. elegans is well suited to HTS as it is easy to grow large isogenic populations and maintain worms in multi-well plates. The use of an
intact organism for the screening process also has many advantages over cell based assays. Most interventions that increase lifespan
also increase resistance to acute stress. This makes it possible to assess the effect of a candidate compound within a few days using
stress resistance assays as a surrogate for lifespan. Here we present different in vivo automated screens for compounds that affect survival
following a stress. We discuss the results of small 2000 compound screens and show that the screens are adaptable to HTS. We discuss
the hurdles to automating whole organism screens and their possible solutions.
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