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Related Concept Videos

DNA Microarrays02:34

DNA Microarrays

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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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Array-on-Array Strategy For Activity-Based Enzyme Profiling.

Brendan Fu-Long Sieow1, Mahesh Uttamchandani2,3,4

  • 1Defence Medical and Environmental Research Institute, DSO National Laboratories, #09-01, 27 Medical Drive, Singapore, 117510, Singapore.

Methods in Molecular Biology (Clifton, N.J.)
|November 23, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces a novel array-on-array method for high-throughput enzymatic activity screening. This approach enables multiplexed testing of various enzymes and samples on a single microarray slide for rapid characterization.

Keywords:
Activity-based screeningArray-on-arrayEnzyme profilingHigh-throughput screeningSmall molecule arrays

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Assay Development

Background:

  • Enzymatic activity screening is crucial for drug discovery and diagnostics.
  • Current methods can be limited in throughput and multiplexing capabilities.
  • Previous droplet-based screening established a foundation for single-enzyme analysis.

Purpose of the Study:

  • To develop a novel array-on-array strategy to enhance enzymatic activity screening throughput.
  • To enable multiplexed analysis of diverse enzymes and mixed samples on a single microarray.
  • To facilitate rapid enzyme characterization using a droplet-based approach.

Main Methods:

  • A two-step spotting process was employed: first, spotting enzymes at varying concentrations, then spotting fluorogenic substrates.
  • The strategy utilizes microarrays for spatial organization of enzyme and substrate droplets.
  • Varying spotting patterns allowed for testing different enzyme classes (proteases, phosphatases, kinases) and substrates.

Main Results:

  • The array-on-array strategy demonstrated enhanced throughput for enzymatic activity screening.
  • Multiplexed screening of multiple enzymes and substrates within a single microarray was achieved.
  • The method allows for simultaneous testing of enzymes and complex biological samples.

Conclusions:

  • The developed array-on-array strategy significantly improves the efficiency of enzymatic screening.
  • This approach offers unprecedented capabilities for multiplexed enzyme testing and characterization.
  • The method builds upon established droplet-based screening for rapid, high-throughput analysis.