LabAutomation 2006 - SLAS

TP35
Julia Michelotti
MDS Sciex
South San Francisco, California
julia.michelotti@sciex.com
Where Laboratory Technologies Emerge and Merge
Co-Author(s)
Roger Tang
Ed Verdonk
Krystal Johnson
Gordon Leung
Ryan McGuinness
MDS Sciex
Identification of the G-protein Coupling Mechanism of GPCRs Using a
Label-free Live-Cell Assay
G-protein coupled receptors (GPCRs) represent the largest target class in drug discovery, and they are important in therapeutic areas
such as heart disease, metabolism and immune disorders. Function is commonly assessed in cells artificially overexpressing the receptor
of interest using fluorescent- or bioluminescent-based probes. With the CellKey system, functional activation of different endogenous
GPCR subtypes can be measured in the same assay format and in their natural setting. The CellKey platform measures changes in
impedance before and after stimulation of endogenous (and transfected, where necessary) receptors and generates unique CDS response
profiles that clearly represent the activation of different GPCR subtype-mediated signal transduction pathways. Here, we demonstrate
these types of data for multiple ligands that activate differently-coupled receptors in several mammalian cell lines (e.g. CHOm1, U-2
OS, HeLa) and primary human osteoblasts. The CellKey assay provides a large advantage over other universal cellular assays in that it
distinguishes between different GPCR subtypes but does not require genetic or chemical manipulation of the cells. Lastly, it is becoming
increasingly apparent that crosstalk between signaling pathways occurs as a result of the normal activation of GPCRs. We show data to
illustrate that the CellKey system can reveal the complex interplay among some GPCR-mediated signaling pathways and that the data
provides information to allow deconvolution of a complex signaling pathway.
TP36
Dana Moss
Corning Incorporated
Corning, New York
mossdd@corning.com
Mannix Aklian, Corning Inc.
Label-Free Detection of Biomolecular Interactions for HTS
A beta version of the 384-well Epic Label-Free Detection System has been used to study several biomolecular assay models. Beyond its
application to direct binding label-free HTS assays on immobilized therapeutic targets, several assays have been derived based on isolated
proteins (kinases, proteases, peptides, and antibodies), membrane preparations (GPCR), cell lysates and whole cells cultured directly on
the Epic microplate. The data generated by some of these assays will be presented.
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