Current Chemical Genomics and Translational Medicine




(Discontinued)

ISSN: 2213-9885 ― Volume 12, 2018

The Use of AlphaScreen Technology in HTS: Current Status



Richard M Eglen*, 1, Terry Reisine1, Philippe Roby2, Nathalie Rouleau2, Chantal Illy2, Roger Bossé2, Martina Bielefeld2
1 President, Molecular Medicine, PerkinElmer Life and Analytical Sciences, 940 Winter St., Waltham, MA 02451-1457, USA
2 PerkinElmer, 1744 Rue William, Suite 600, Montreal Qc H3J 1R4, Canada

Abstract

AlphaScreen (Amplified Luminescent Proximity Homogeneous Assay Screen) is versatile assay technology developed to measuring analytes using a homogenous protocol. This technology is an example of a bead-based proximity assay and was developed from a diagnostic assay technology known as LOCI (Luminescent Oxygen Channeling Assay). Here, singlet oxygen molecules, generated by high energy irradiation of Donor beads, travel over a constrained distance (approx. 200 nm) to Acceptor beads. This results in excitation of a cascading series of chemical reactions, ultimately causing generation of a chemiluminescent signal.

In the past decade, a wide variety of applications has been reported, ranging from detection of analytes involved in cell signaling, including protein:protein, protein:peptide, protein:small molecule or peptide:peptide interactions. Numerous homogeneous HTS-optimized assays have been reported using the approach, including generation of second messengers (such as accumulation of cyclic AMP, cyclic GMP, inositol [1, 4, 5] trisphosphate or phosphorylated ERK) from liganded GPCRs or tyrosine kinase receptors, post-translational modification of proteins (such as proteolytic cleavage, phosphorylation, ubiquination and sumoylation) as well as protein-protein and protein-nucleic acid interactions.

Recently, the basic AlphaScreen technology was extended in that the chemistry of the Acceptor bead was modified such that emitted light is more intense and spectrally defined, thereby markedly reducing interference from biological fluid matrices (such as trace hemolysis in serum and plasma). In this format, referred to as AlphaLISA, it provides an alternative technology to classical ELISA assays and is suitable for high throughput automated fluid dispensing and detection systems.

Collectively, AlphaScreen and AlphaLISA technologies provide a facile assay platform with which one can quantitate complex cellular processes using simple no-wash microtiter plate based assays. They provide the means by which large compound libraries can be screened in a high throughput fashion at a diverse range of therapeutically important targets, often not readily undertaken using other homogeneous assay technologies. This review assesses the current status of the technology in drug discovery, in general, and high throughput screening (HTS), in particular.


Article Information


Identifiers and Pagination:

Year: 2008
Volume: 1
First Page: 2
Last Page: 10
Publisher Id: CCGTM-1-2
DOI: 10.2174/1875397300801010002

Article History:

Received Date: 20/12/2007
Revision Received Date: 13/1/2008
Acceptance Date: 19/1/2008
Electronic publication date: 25/2/2008
Collection year: 2008

©2008 Bentham Science Publishers Ltd.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.5/), which permits unrestrictive use, distribution, and reproduction in any medium, provided the original work is properly cited.


* Address correspondence to this author at Molecular Medicine, PerkinElmer Life and Analytical Sciences, 940 Winter St., Waltham, MA 02451-1457, USA; Tel: 781-663-5599; Fax: 781-663-5984; E-mail: richard.eglen@perkinelmer.com


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