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High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents (HPHC)
Published on: May 10, 2016
The use of AlphaScreen technology in HTS: current status
Richard M Eglen1, Terry Reisine, Philippe Roby
1President, Molecular Medicine, PerkinElmer Life and Analytical Sciences, 940 Winter St., Waltham, MA 02451-1457, USA. richard.eglen@perkinelmer.com
Current Chemical Genomics
|February 18, 2010
Summary
AlphaScreen and AlphaLISA are bead-based proximity assay technologies for measuring analytes. These homogeneous, no-wash assays facilitate high-throughput screening for drug discovery by detecting complex cellular processes.
Area of Science:
- Biochemistry
- Assay Technology
- Drug Discovery
Background:
- AlphaScreen is a bead-based proximity assay utilizing singlet oxygen transfer for chemiluminescent signal generation.
- It is derived from Luminescent Oxygen Channeling Assay (LOCI) technology.
- Applications span cell signaling, protein interactions, and post-translational modifications.
Purpose of the Study:
- To review the current status of AlphaScreen and AlphaLISA technologies in drug discovery and high-throughput screening (HTS).
- To highlight their versatility in quantitating complex cellular processes.
- To emphasize their utility in screening large compound libraries against therapeutic targets.
Main Methods:
- Utilizes Donor and Acceptor beads for proximity-based detection.
- Generates a chemiluminescent signal through a cascade of reactions initiated by singlet oxygen.
- AlphaLISA offers enhanced signal intensity and spectral definition, reducing matrix interference.
Main Results:
- Demonstrated wide-ranging applications in detecting protein:protein, protein:peptide, and protein:small molecule interactions.
- Successfully employed in HTS assays for second messenger generation and protein modifications.
- AlphaLISA provides a robust alternative to ELISA for automated HTS systems.
Conclusions:
- AlphaScreen and AlphaLISA are facile, no-wash assay platforms for quantitating cellular processes.
- These technologies enable high-throughput screening of compound libraries against diverse therapeutic targets.
- They offer advantages over other homogeneous assay technologies for drug discovery efforts.
