Detecting molecular interactions by fluorescence resonance energy transfer on a solid-phase support
The present disclosure features a variety of compositions, kits, and methods that are useful for, inter alia, detecting and/or analyzing an interaction between two molecules, a target molecule and a target-specific binding agent which can be, e.g., proteins, nucleic acids, saccharides or polysaccharides, small molecules, or combinations of any of the foregoing. The compositions, kits, and methods can also be used, e.g., to detect the presence or absence of an enzymatic activity (e.g., a kinase activity, a protease activity, or a phosphatase activity) in a sample; to identify a compound that modulates an interaction between two molecules; or to identify compounds that modulate the activity of an enzyme.
1. A composition comprising:
a first solid-phase support comprising a target-selective binding agent and a first member of a Fluorescence Resonance Energy Transfer (FRET) pair; and
a target molecule associated with a second member of the FRET pair, wherein the target molecule is not associated with a solid-phase support;
wherein the first solid-phase support comprises phthalocyanine and thioxene;
wherein the first member of the FRET pair is a lanthanide chelate; and
wherein the target-selective binding agent comprises deferoxamine-Fe 3+ and preferentially binds to a target molecule that comprises a phosphomonoester moiety as compared to a target molecule that does not comprise a phosphomonoester moiety.
2. The composition of claim 1 , wherein the first solid-phase support is an assay plate.
3. The composition of claim 1 , wherein the first solid-phase support is a particle.
4. The composition of claim 1 , wherein the lanthanide chelate comprises a lanthanide selected from the group consisting of europium, terbium, samarium, and dysprosium.
5. The composition of claim 1 , wherein the composition comprises at least two first solid-phase supports, each first solid-phase support comprising a different target-selective binding agent.
6. The composition of claim 1 , wherein the first solid-phase support is a particle having a diameter between about 100 nm and 1000 nm.
7. The composition of claim 1 , wherein the first solid-phase support is a particle having a diameter between about 100 nm and 400 nm.
8. The composition of claim 1 , wherein the composition is in suspension or is lyophilized.
9. The composition of claim 1 , wherein the composition further comprises an aqueous buffer in which the composition forms a suspension or solution.
10. The composition of claim 1 , wherein the target molecule comprises one or more of a polypeptide, a nucleic acid, a carbohydrate, and a small molecule.
11. The composition of claim 10 , wherein the target molecule comprises a polypeptide.
12. The composition of claim 10 , wherein the target molecule comprises a nucleic acid.
13. The composition of claim 10 , wherein the target molecule comprises a carbohydrate.
14. The composition of claim 10 , wherein the target molecule comprises a small molecule.