Protease-responsive peptide biosensors and methods for analyte detection
AssigneeAgency for Science, Technology and Research
InventorsSaurabh Rajendra Nirantar, Farid John Ghadessy
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Abstract
The present invention relates to peptide biosensors comprising (a) a protease recognition site; (b) an analyte binding site; and (c) a signaling moiety that can produce a detectable signal upon cleavage of the protease recognition site by a protease. Further encompassed are detection reagents that comprise said peptide biosensors in combination with a protease and methods for detecting the presence of an analyte molecule or screening of candidate compounds that modulate the binding of an analyte to a binding partner of the analyte.
Claims20
1-27. (canceled)
Peptide biosensor for the detection of an analyte, wherein the peptide biosensor comprises
- (a) a protease recognition site;
- (b) an analyte binding site;
- and
- (c) a signaling moiety that can produce a detectable signal upon cleavage of the protease recognition site by a protease,
- wherein the protease recognition site and the analyte binding site are positioned relative to each other such that binding of the analyte to the analyte binding site reduces or prevents binding of the protease to the protease recognition site, or wherein the protease recognition site is an attenuated protease recognition site.
Detection reagent for the detection of an analyte, wherein the detection reagent comprises
- (1):
- (a) a peptide biosensor comprising
- a protease recognition site;
- an analyte binding site;
- and
- a signaling moiety that can produce a detectable signal upon cleavage of the protease recognition site by a protease,
- wherein the protease recognition site and the analyte binding site are positioned relative to each other such that binding of the analyte to the analyte binding site reduces or prevents binding of the protease to the protease recognition site, or wherein the protease recognition site is an attenuated protease recognition site;
- and
- (b) a protease capable of binding to and cleaving the protease recognition site;
- or
- (2):
- (A) a solid substrate comprising immobilized thereon:
- (a) a peptide biosensor comprising
- (i) a protease recognition site;
- and
- (ii) a signaling moiety,
- wherein the peptide biosensor is immobilized such that upon cleavage of the protease recognition site by a protease, the signaling moiety gets released from the substrate.
- (b) a first (poly)peptide capable of binding the analyte, wherein said first (poly)peptide is immobilized on the substrate in proximity of the peptide biosensor;
- and
- (B) a protease coupled to a second (poly)peptide capable of binding the analyte, wherein both the first (poly)peptide immobilized on the substrate and the second (poly)peptide coupled to the protease can simultaneously bind to the analyte.
Dependencies are inferred from the claim text.