Diagnostic methods and compositions
AssigneeThe Regents of the University of California
InventorVictor Emilio MUÑOZ VAN DEN EYNDE
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Abstract
The present disclosure provides methods for the identification and quantitation of targets (e.g., biological targets such as cells and viruses) using molecules comprising a binding moiety that binds to the target and a fluorescent moiety whose fluorescence properties are altered when the binding moiety binds to the target. A plurality of said molecules are specifically arranged in structures that resemble the size and shape of a cell to maximize affinity and sensitivity.
Claims29
A composition, comprising:
- an engineered protein including at least one amino acid sequence that binds to at least one target molecule;
- a donor moiety and an acceptor moiety fused to the engineered protein;
- wherein the engineered protein undergoes a first conformation change to unfold in the absence of the target molecule and undergoes a second conformational change to fold upon binding to the target molecule using a fraction of a binding free energy provided by binding of the engineered protein to the target molecule or wherein the engineered protein undergoes the second conformation change to fold in the absence of the target molecule and undergoes the first conformation change to unfold upon binding to the target molecule;
- and
- wherein responsive to the first or the second conformational change, an energy transfer or an electron transfer occurs between the donor moiety and the acceptor moiety.
A composition for detection of a target, comprising:
- a downhill folding protein including a binding moiety that binds to a molecule on the surface of the target;
- and
- a fluorescent moiety fused to the downhill folding protein, the fluorescent moiety having a first emission property when the binding moiety is not bound to the target, and a second emission property when the binding moiety is bound to the target,
- wherein the first and second emission properties are selected from the group consisting of one or both of fluorescence intensity and emission wavelength;
- and
- wherein the downhill folding protein undergoes a conformational change responsive to binding to the target
A composition for detecting a target, the composition comprising:
- a plurality of responsive proteins;
- wherein the plurality of responsive proteins are assembled on to a plurality of substrates through engineered multivalent adhesion at a desired density per substrate to generate a host cell decoy for the target.
Dependencies are inferred from the claim text.