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A Real-Time Fluorescence Assay for Protease Activity and Inhibitor Screening Based on the Aggregation-Caused Quenching of a Perylene Probe

A Real-Time Fluorescence Assay for Protease Activity and Inhibitor Screening Based on the Aggregation-Caused Quenching of a Perylene Probe

Yan Wang, Zhifang Zhang, Ya Zhang, Cong Yu

Luminescence. 2018 Jun;33(4):790-796.

PMID: 29607616

Abstract:

We have established a real-time and label-free fluorescence turn-on strategy for protease activity detection and inhibitor screening via peptide-induced aggregation-caused quenching of a perylene probe. Because of electrostatic interactions and high hydrophilicity, poly-l-glutamic acid sodium salt (PGA; a negatively charged peptide) could induce aggregation of a positively charged perylene probe (probe 1) and the monomer fluorescence of probe 1 was effectively quenched. After a protease was added, PGA was enzymatically hydrolyzed into small fragments and probe 1 disaggregated. The fluorescence recovery of probe 1 was found to be proportional to the concentration of protease in the range from 0 to 1 mU/ml. The detection limit was down to 0.1 mU/ml. In the presence of a protease inhibitor, protease activity was inhibited and fluorescence recovery reduced. Moreover, we demonstrated the potential application of our method in a complex mixture sample including 1% human serum. Our method is simple, fast and cost effective.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP26247790	Poly-L-glutamic acid sodium salt		26247-79-0	Price

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