Related Experiment Video
Updated: Jul 3, 2026
11:57
Fabrication and Use of MicroEnvironment microArrays (MEArrays)
Published on: October 11, 2012
Enzyme microarrays assembled by acoustic dispensing technology
1Penn Center for Molecular Discovery, Institute for Medicine and Engineering, Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, PA 19104, USA.
Analytical Biochemistry
|July 12, 2008
Summary
Acoustic dispensing miniaturizes bioassays to nanoliter volumes for high-throughput screening. This cost-effective method enables precise protein kinase assays and inhibitor screening, reducing reagent use.
Area of Science:
- Biochemistry
- Assay Development
- High-Throughput Screening
Background:
- Miniaturizing bioassays reduces reagent consumption, crucial for expensive or difficult-to-handle substances.
- Acoustic dispensing technology enables precise nanodroplet formation for miniaturized assays.
Purpose of the Study:
- To develop and validate a miniaturized protein kinase assay using acoustic dispensing.
- To assess the effectiveness of acoustic dispensing for high-throughput screening of enzyme inhibitors.
Main Methods:
- Nanoliter droplets (40 nL) containing ATP and reaction buffer were dispensed onto glass slides.
- Pim1 kinase reactions were initiated by dispensing inhibitor, enzyme, and peptide substrate in 4 nL volumes.
- Phosphor imaging quantified phosphorylated substrate after blotting to phosphocellulose membranes.
Main Results:
- The assay successfully demonstrated increasing inhibition of Pim1 kinase with higher doses of a ruthenium complex inhibitor (0.75–3 microM).
- High signal-to-background ratios (up to 165) and acceptable assay variation (CV ~20%) were achieved.
- Acoustic dispensing of buffers showed excellent precision (CV < 5%).
Conclusions:
- Microarrays assembled by acoustic dispensing are effective for miniaturized protein kinase assays.
- This technology offers a promising, cost-effective tool for protein assay development and inhibitor screening.
- The method facilitates high-throughput screening with reduced reagent usage.
