It is herein disclosed a new methodol. to detect an enzyme, in particular a protease (i.e. a protein capable of hydrolyzing peptide ligands). The detn. is based on the activation of colloidal gold nanoparticles with catalytic properties via enzyme hydrolysis, operated by the enzyme to be detd., of a peptide sequence acting as inhibitor for said Au nanoparticles. Following from hydrolysis, the Au nanoparticles' catalytic activity is restored by detg. the prodn. of great quantities of a "reporter" mol. via spectrophotometry. The method can be made selective with regards to the enzyme to be detd. by using a specific peptide sequence. The proteases of interest for the method are subtilisin, caspases, glutamate carboxy peptidase II, granzyme and thermolysin.
Detection of a proteolytic enzyme by means of gold nanoparticles-catalyzed amplification of a signal
PRINS, LEONARD JAN;SCRIMIN, PAOLO MARIA;BONOMI, RENATO
2012
Abstract
It is herein disclosed a new methodol. to detect an enzyme, in particular a protease (i.e. a protein capable of hydrolyzing peptide ligands). The detn. is based on the activation of colloidal gold nanoparticles with catalytic properties via enzyme hydrolysis, operated by the enzyme to be detd., of a peptide sequence acting as inhibitor for said Au nanoparticles. Following from hydrolysis, the Au nanoparticles' catalytic activity is restored by detg. the prodn. of great quantities of a "reporter" mol. via spectrophotometry. The method can be made selective with regards to the enzyme to be detd. by using a specific peptide sequence. The proteases of interest for the method are subtilisin, caspases, glutamate carboxy peptidase II, granzyme and thermolysin.Pubblicazioni consigliate
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