LabAutomation 2006 - SLAS

MP29
Jon Curtis
GSK
Stevenage, Herfordshire, United Kingdom
jpc67195@gsk.com
Where Laboratory Technologies Emerge and Merge
Co-Author(s)
Zoe Blaxill, Suzanne Baddeley, Liz Clark, Jim Chan, Jim Laugharn,
Covaris
Phil Robinson, Kbioscience
The Use of Adaptive Focused Acoustic Technology to Improve uHTS Assay
Performance and Compound Dissolution.
Compound Management and high throughput screening can both be impacted by incomplete compound dissolution as well as precipitation,
the latter occurring on storage or after addition of aqueous buffer. To date, mixing and dissolution have been performed using vortexing,
sonication or centrifugation. However, all these methods have drawbacks.
Mixing in low volume 384 and 1536 plate formats also presents difficulties in overcoming high surface tensions and slow diffusion kinetics.
High frequency focused acoustics has demonstrated effective mixing in 1536 formats in multiple assay types including SPA bead based
assays, which are particularly prone to settling thus reducing the efficiency of the assay.
Adaptive focused acoustics has been shown to significantly improve the Z’ value, lower standard deviation, accelerate assays and reduce
compound precipitate in aqueous. All these factors increase positive confidence by reducing false positives which lessen the requirement
for retests.
High Frequency Adaptive Acoustic technology from Covaris is a patented non-contact, isothermal technology capable of rapid mixing
in all common types of tubes, vials and high density microtitre plate formats. We will describe the acoustic technology and how it has
been evaluated within GlaxoSmithKline. Data will be presented from different areas of Compound Management and ultra high throughput
screening (uHTS), including compound dissolution, investigation of compound degradation and improved assay performance assay data.
Sample formats covered will include 2D coded tubes, 4ml vials, 96, 384 and 1536 well microtitre plates.
MP30
Teresa Damico
Wayne State University
Detroit, Michigan
tdamico@chem.wayne.edu
Co-Author(s)
Paul Root
Dana M. Spence
Wayne State University
Detection of Endothelial Cell Derived Nitric Oxide Using a Microfluidic Device
With the advancement of immobilizing bovine pulmonary artery endothelial cells (bPAECs) in confluent layers within poly(dimethylsiloxane)
based micro-chip channels, a need has arisen to monitor certain physiological occurrences, such as signal transmission by nitric oxide, in
the channel using methods other than amperometry. One such method that has become of interest in recent years is the use of fluorescent
indicators, and of particular importance are diaminofluorescein (DAF) derivatives. Here, a difluorinated derivative, DAF-FM, was employed
to conduct studies in which adenosine tri-phosphate (ATP) was used as a stimulus to observe nitric oxide production in bPAECs. We were
also able to utilize L-NAME, a known competitive inhibitor of nitric oxide synthase, to verify that the resultant NO production was the result
of ATP stimulation. When combined with microfluidic methods, such a system has the potential for high-throughput drug efficacy studies.
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