This technology is a rapid, simple and sensitive method for detecting the binding or unbinding of biomolecules using a silicon wafer. It is a novel method for detecting when two biomolecules unbind. The target biomolecule is placed on the silicon wafer, and then the partner biomolecules is fluorescently labeled and allowed to bind to target protein. At this point, the fluorescent label is quenched as it is brought into proximity of the silicon substrate. The technology works best for detecting disassociation or unbinding, such as screening for drugs that interfere with protein-protein binding.
Compared to quenching by the common technique of fluorescence resonance energy transfer (FRET), the technology yields much larger quenching (signal/background ratio); provides robust signals even on the largest biomolecules complexes; and simplifies labeling since only one protein must be labeled (compared with two labels needed for FRET). Also, position or rigidity of the labeling site is relatively unimportant compared with polarized fluorescence assays. Because silicon is used, photolithographic techniques can potentially be used to miniaturize and integrate excitation, detection, and sample handling. Finally, the technology is fluorescence-based so it is highly sensitive and safe without the need for radioactivity.
Applications
This technology would be highly useful in therapeutic drug screening, and should be of interest to major pharmaceutical companies. High-throughput screening for drug discovery Works best for measuring disassociation/unbinding
Benefits
- Long-quenching range: more efficient at energy transfer than the standard fluoresce resonance energy transfer (FRET), thus larger complexes still receive energy transfer
- Simplified Labeling: acceptor protein does not have to be labeled and is less critical where dye is attached to donor
- Scalable: Possible to miniaturize technology Safe: Technology does not use radioactivity