title: Peptide-induced fluorescence quenching of conjugated polyelectrolyte for label-free, ultrasensitive and selective assay of protease activity.
abstract: We report here a label-free method for ultrasensitive and selective assay of protease activity based on the peptide-induced fluorescence quenching of conjugated polyelectrolyte (PPESO(3)). It is very interesting to find that there is a critical length of oligo-polyarginine (i.e., Arg(5)) below which 1) the quenching efficiency of PPESO(3) is sharply decreased, and more importantly, 2) the trypsin-catalyzed hydrolysis is greatly slowed down. This opens good opportunities for not only the ultrasensitive assay of trypsin, but the specific detection of other proteases if carefully designing an appropriate peptide length and the cleavage site. Herein, the enzyme selected as a proof of concept is chymotrypsin. Due to the essence that any cleavage of the designed peptide probes will result in a notable decrease or even a complete loss of their capability to quench the emission of PPESO(3), the limits of detection for trypsin and chymotrypsin have been found as low as 0.25 ng/mL (11 pM) and 0.15 ng/mL (6 pM), respectively. Both are superior to those of most previous methods by 1-2 orders or higher.
authors.affiliation: Research Center for Analytical Instrumentation, Institute of Cyber-Systems and Control, State Key Laboratory of Industrial Control Technology, Zhejiang University, Hangzhou, PR China.
authors.forename: Hongliang
authors.initials: H
authors.lastname: Fan
authors.forename: Xinhang
authors.initials: X
authors.lastname: Jiang
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.forename: Qinhan
authors.initials: Q
authors.lastname: Jin
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002918:Chymotrypsin / Q000737:chemistry / Q000302:isolation & purification*; D005453:Fluorescence; D006868:Hydrolysis; D009842:Oligopeptides / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000737:chemistry / Q000302:isolation & purification*; D010455:Peptides / Q000737:chemistry*; D012680:Sensitivity and Specificity
chemical_list: D009842:Oligopeptides; D010455:Peptides; C015462:polyarginine; D010447:Peptide Hydrolases; D002918:Chymotrypsin