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Putting thought to paper: a microARCS protease screen.
Duncan R Groebe1, Mary L Maus, Terry Pederson
1Biological Screening, Abbott Laboratories, Abbott Park, IL, USA. duke.groebe@abbott.com
Journal of Biomolecular Screening
|January 9, 2004
Summary
Micro-arrayed compound screening (microARCS) was enhanced using blotter paper for substrate delivery, achieving over 200,000 tests per hour. This method successfully identified small molecule enzyme inhibitors.
Area of Science:
- Biochemistry
- Assay Development
- High-Throughput Screening
Background:
- Micro-arrayed compound screening (microARCS) utilizes agarose gels as reaction vessels for compound screening.
- Agarose gel matrix regulates reactant diffusion for enzymatic reactions.
Purpose of the Study:
- To increase throughput, reduce effort, and lower costs in microARCS.
- To adapt microARCS for high-throughput screening by replacing a second agarose gel with blotter paper.
Main Methods:
- Developed a microARCS assay using blotter paper for substrate introduction to a target enzyme in an agarose gel.
- Validated the assay with a protease assay, achieving 200,000 tests per hour.
- Confirmed IC50 values in a 96-well plate format for identified hits.
Main Results:
- The blotter paper matrix allowed substrate diffusion into the enzyme gel.
- Achieved a throughput of over 200,000 tests per hour.
- Identified several small molecule inhibitors of the target enzyme.
Conclusions:
- Blotter paper is an effective and efficient substrate delivery method for microARCS.
- The modified microARCS format significantly enhances screening throughput.
- This method facilitates the discovery of novel small molecule inhibitors.