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High-Pressure NMR Experiments for Detecting Protein Low-Lying Conformational States
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Second-harmonic generation-based methods to detect and characterize ligand-induced RNA conformational changes.

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Second-harmonic generation (SHG) technology enables high-throughput screening of biomolecular conformational changes for drug discovery. Optimized RNA assays using SHG provide robust measurements for identifying potential drug candidates.

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Area of Science:

  • Biophysics
  • Biochemistry
  • Drug Discovery

Background:

  • Second-harmonic generation (SHG) is a biophysical technique used to detect conformational changes in biomolecules.
  • The Biodesy Delta platform utilizes SHG for high-throughput screening of proteins and oligonucleotides.
  • This technology supports drug discovery by monitoring molecular interactions.

Purpose of the Study:

  • To provide an overview of SHG technology for biomolecular screening.
  • To detail protocols for optimizing SHG assays on RNA targets.
  • To present practical considerations and a case study for developing robust SHG measurements.

Main Methods:

  • Optimization of a theophylline aptamer SHG assay.
  • Testing for specific conjugate tethering to surfaces.
  • Evaluating tool compound response stability, reversibility, and concentration-dependence.

Main Results:

  • An improved SHG assay for the theophylline aptamer was developed.
  • Maximized assay window between positive (theophylline) and negative (caffeine) controls.
  • Demonstrated practical considerations for robust RNA SHG assay development.

Conclusions:

  • Optimized SHG assays are crucial for effective drug discovery screening.
  • Robust assay development involves careful consideration of target immobilization and compound response.
  • The enhanced theophylline aptamer assay is suitable for high-throughput screening applications.