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Related Experiment Videos

Label-free high-throughput functional lytic assays.

Shawn M O'Malley1, Xinying Xie, Anthony G Frutos

  • 1Corning Incorporated, Corning, NY 14831, USA. omalleysm@corning.com

Journal of Biomolecular Screening
|December 19, 2006
PubMed
Summary

This study demonstrates label-free biosensors for enzyme activity screening. These refractive index-based sensors accurately measure biomolecular degradation, enabling robust high-throughput screening of functional modulators.

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Area of Science:

  • Biochemistry
  • Biotechnology
  • Biosensor Technology

Background:

  • Refractive index-sensitive resonant waveguide grating biosensors offer label-free assay capabilities for biomolecular interactions.
  • Enzymatic degradation assays are crucial for identifying functional modulators, but often require complex labeling techniques.

Purpose of the Study:

  • To evaluate label-free enzymatic degradation assays using resonant waveguide grating biosensors for screening functional lytic modulators.
  • To determine the substrate and protease specificity of label-free enzymatic digestion.
  • To assess the feasibility of this biosensor platform for high-throughput screening (HTS).

Main Methods:

  • Covalent immobilization of biomolecular substrates via amine groups onto biosensors.
  • Measurement of protein substrate digestion using the Corning Epic System.

Related Experiment Videos

  • Determination of lytic inhibitor properties and IC50 values for proteases and dextranase.

    • Main Results:

    • Label-free digestion profiles exhibited substrate and protease specificity.
    • The biosensor platform demonstrated robust performance with Z' values consistently above 0.5.
    • IC50 values for inhibitors correlated well with previously reported data.

    Conclusions:

    • Resonant waveguide grating biosensors provide a robust and label-free method for assaying enzymatic degradation.
    • This technology is suitable for high-throughput screening of functional lytic modulators.
    • The specificity and accuracy of the assay support its application in drug discovery and enzyme characterization.