Patent ID: 9249451
Filing Date: 2016-02-02
CPC Classification: C07K,C12Q,G01N

Claim Text:
1. A method of improving sensitivity of energy transfer between a donor label and an acceptor label, comprising: providing an initial construct comprising the donor label and the acceptor label physically coupled through a linker, the linker comprising a cleavage site sequence, wherein the cleavage site sequence comprises a SNARE protein, or a fragment or mutein thereof selected to be susceptible to cleavage by a botulinum neurotoxin; incorporating into the linker, in addition to the cleavage site sequence, a flexible peptide spacer selected to increase electronic coupling between the donor label and the acceptor label by facilitating an electronic hop between the donor label and the acceptor label between at least one of the donor label and the cleavage site sequence and the acceptor label and the cleavage site sequence to provide an improved construct; and characterizing a FRET emission from the improved construct is; wherein the improved construct demonstrates improved sensitivity in a FRET-based assay directed to a botulinum neurotoxin relative to the initial construct.