Electrochemical assay for the detection of enzymes

摘要

The invention relates to novel compositions and methods for the detection of enzymes using the nuclear reorganization energy, λ, of an electron transfer process.

主权项

1. A method for detecting for the presence of a protease in a test sample, said method comprising:
(a) adding a test sample to a solid support comprising an electrode comprising:
(i) a self-assembled monolayer (SAM);(ii) a covalently attached electroactive active moiety (EAM) comprising a transition metal complex with an E0, wherein said EAM comprises a ferrocene; and(iii) a plurality of proteins attached to said electrode, wherein said proteins are arranged so that the EAM is at least partially shielded by the proteins from exposure to a solution and comprise a cleavage site of said protease, wherein the cleavage site is near the height of said EAM; (b) cleaving a plurality of said proteins with said protease, if said protease is present, such that when the proteins are cleaved at the cleavage site, the EAM is exposed to the solution; and (c) determining the presence of said protease by measuring a change of said E0.