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Detection of protein-small molecule binding using a self-referencing external cavity laser biosensor
Summary
We developed a novel self-referencing external cavity laser (ECL) biosensor for label-free detection of protein-small molecule interactions. This high-resolution biosensor accurately identifies binding affinities, enabling efficient drug discovery screening.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Biotechnology
Background:
- Label-free optical biosensors are crucial for high-throughput screening of protein-small molecule binding.
- Detecting weak binding signals amidst experimental noise remains a significant challenge.
Purpose of the Study:
- To introduce a novel self-referencing external cavity laser (ECL) biosensor for enhanced sensitivity and resolution in binding assays.
- To demonstrate the biosensor's capability for accurate detection of protein-small molecule interactions across a range of affinities.
Main Methods:
- Development of a self-referencing external cavity laser (ECL) biosensor.
- Utilizing sub-picometer wavelength accuracy to eliminate thermal noise.
- Immobilization of protein targets for binding assays with small molecules.
Main Results:
- Achieved high resolution and sensitivity in label-free detection of binding interactions.
- Successfully detected binding affinities and inhibition constants from sub-nanomolar to low micromolar range.
- Demonstrated robust detection in the presence of interfering compounds, enhancing screening potential.
Conclusions:
- The self-referencing ECL biosensor offers a powerful tool for sensitive and accurate label-free screening of protein-small molecule interactions.
- This approach can significantly improve the throughput and reliability of drug discovery efforts, as exemplified by carbonic anhydrase II inhibitor screening.
