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PerkinElmer's
Toolbox for High-Content Analysis
Richard
Eglen, Vice President Discovery and Research Reagents
for PerkinElmer, talked to Pharma
DD about their
latest tools for high-content cell-based screening.
On
the drug discovery side, PerkinElmer has licensed
access to Luminex’s xMAP technology, which enables
us to offer multiplex bead-based assay systems. These
systems allow the measurement, with high sensitivity,
of cell-based responses that give off a luminescent
signal. The xMAP platform is being used for primary
screening in terms of cell-based assays, as well as
secondary screening with some high-content assays.
PerkinElmer is strongly interested in developing more
luminescent reagent systems. Our customers have a high
level of interest in measuring luminescence in assays,
partly because of its high level of sensitivity, but
also because when using luminescence there are fewer
interferences from the screening lab introduced to the
sample.
Screen
for Difficult Protein Targets
In
addition to that, PerkinElmer’s own product line is
the highly sensitive AlphaScreen assay technology,
which is used for high-throughput screening and is
scalable to 384- and 1536-well microplate formats. The
AlphaScreen technology has also been adapted so that
ELISA-based assays can be performed without having to
do a wash and separation step. ELISA is a laborious
and time-consuming process due to its multiple washing
steps.
AlphaScreen
provides a homogeneous and sensitive alternative assay
to ELISA format. In that case, we’re looking less at
cell responses and are using it more to measure
secreted proteins and cytokines. Some of these systems
also measure key biomarkers of cellular toxicity, so
it is possible to take some of these technologies into
the ADME/Tox area too.
PerkinElmer
has an agreement with TGR BioSciences, who has adapted
the AlphaScreen platform to measure phosphorylated
proteins activated by cell-signaling pathways. The
combination of AlphaScreen with TGR’s SureFire
cellular kinase assay kits permits primary and
secondary screening of previously difficult targets
through homogenous non-radioactive measurement of
phosphorylated full-length proteins. The kits permit
researchers to screen a number of key modulators of
pathway activation in whole cells. The technology
platform can be used with many different cell lines,
including primary cells, cells over-expressing cloned
targets, or cells expressing physiologically relevant
endogenous levels of kinase or receptor target.
The
AlphaScreen SureFire
ERK assay, which was the first product to be launched
from the collaboration with TGR, expanded our offering
in the field of cell-based functional G-Protein
Coupled Receptor (GPCR) screening. It has been
successfully used to screen for therapeutic modulators
of GPCR and Receptor Tyrosine Kinase (RTK) targets.
This initial assay has now been extended to a series
of ten assays looking at key proteins that are
phosphorylated as a particular pathway is activated.
Perform Quantitative Proteomics
Some
of PerkinElmer’s reagents and products are geared
toward genomic platforms, while others speak to
proteomic platforms. Proteomics is very important in
drug discovery, especially in terms of studying kinase-activated
pathways. To that point, we acquired Agilix
Corporation in March. They developed a multiplex
protein labeling technology using isobaric mass tags,
allowing for the simultaneous quantification of
proteins from multiple samples. For example, the
proteins could be specifically tagged to measure their
phosphorylation. It is a mass spectrometry-based
approach, allowing for the quantitative measurement of
the nature and the amount of the protein that is
phosphorylated. The approach determines the relative
expression level of individual proteins from multiple
samples with a high degree of precision.
At
last month’s Human Proteome Organization (HUPO)
World Congress, PerkinElmer launched three new
proteomic research technologies: ExacTag mass tags,
Phos-tools, and Affinity-tools. The trio is compatible
with mass spectrometry work flows.
ExacTag
mass tags are a family of thiol and amine reactive
isobaric mass tags that offer flexibility for the
quantification of protein expression of up to ten
samples simultaneously, enabling analysis of protein
expression pharmacodynamics.
Phos-tools
utilize unique chemistries to provide a high binding
selectivity for phosphorylated tyrosine, serine, and
threonine residues.
The Phos-tools isolate and detect
phosphorylation events with enhanced sensitivity and
selectivity over current IMAC technology.
Affinity-tools
are based upon conventional immuno- and biotin-binding
protein capture strategies, allowing for the selective
isolation of proteins of interest and sensitive
detection by gel resolution or by mass spectrometry.
Tools for Gene Expression Validation and
Cytogenetics
In
May, PerkinElmer acquired Spectral Genomics, which has
an Array CGH (Comparative Genome Hybridization)
technology that enables rapid surveys of the whole
genome at high resolution and can identify the exact
location of any chromosomal abnormalities (e.g.
deletions and amplifications) that can cause an
increased risk of genetic disease. The whole area of
CGH on an array basis is one that is growing very
quickly.
The
acquisition of Spectral Genomics has really propelled
PerkinElmer into that space. Last month, at the
American Society of Human Genetics meeting in
New Orleans
, we launched both the Comprehensive Solution (CS)
Autoplex Gene Expression Platform and the Spectral
Genomics Array CGH Platform.
The CS
Autoplex Platform contains two products for gene
expression validation--a custom gene expression panel
kit and a labeling kit, both with analysis software.
The CS Autoplex Custom Gene Expression Panel Kit
enables customization of candidate gene panels for the
high-throughput validation of large sample sets. These
kits are built upon the Luminex xMAP technology. The
CS Autoplex Workstation completes the CS Platform by
offering automated multiplex technology for assay set
up, sample processing, and analysis for high-content
xMAP technology.
The
Spectral Genomics Array CGH Platform offers a
Bacterial Artificial Chromosome (BAC)-based solution
consisting of the SpectralChip 2600 array, the
Constitutional Chip 3.0 array, the ScanArray System,
labeling reagents, and SpectralWare Analysis Software.
The platform, which can be used for discovery and
investigational purposes, can generate results in 24
hours versus the industry standard of five to 20 days
in the lab, which is common for standard karyotyping.
For
discovery, the SpectralChip 2600 array is the only
complete BAC array platform for genome profiling. For
cytogenetics, the Constitutional Chip 3.0 array is
being evaluated for coverage of 42 disorders and 41
subtelomeric regions as an alternative to current
Fluorescence In Situ Hybridization (FISH) technology.
For
more information on PerkinElmer’s offerings, visit www.perkinelmer.com.
Copyright 2006, Cambridge
Healthtech Institute. All Rights Reserved.
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