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Open AccessArticle

Label-Free Fluorescent Detection of Trypsin Activity Based on DNA-Stabilized Silver Nanocluster-Peptide Conjugates

by Cai-Xia Zhuo 1,2,†, Li-Hui Wang 1,2,†, Jing-Jing Feng 1,2 and Yao-Dong Zhang 1,2,*,†
1
Key Laboratory of Applied Surface and Colloid Chemistry of Ministry of Education, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi’an 710062, China
2
Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi’an 710062, China
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editors: Jong Seung Kim and Min Hee Lee
Sensors 2016, 16(11), 1477; https://doi.org/10.3390/s16111477
Received: 27 June 2016 / Revised: 2 August 2016 / Accepted: 2 August 2016 / Published: 9 November 2016
(This article belongs to the Special Issue Colorimetric and Fluorescent Sensor)
Trypsin is important during the regulation of pancreatic exocrine function. The detection of trypsin activity is currently limited because of the need for the substrate to be labeled with a fluorescent tag. A label-free fluorescent method has been developed to monitor trypsin activity. The designed peptide probe consists of six arginine molecules and a cysteine terminus and can be conjugated to DNA-stabilized silver nanoclusters (DNA-AgNCs) by Ag-S bonding to enhance fluorescence. The peptide probe can also be adsorbed to the surface of graphene oxide (GO), thus resulting in the fluorescence quenching of DNA-AgNCs-peptide conjugate because of Förster resonance energy transfer. Once trypsin had degraded the peptide probe into amino acid residues, the DNA-AgNCs were released from the surface of GO, and the enhanced fluorescence of DNA-AgNCs was restored. Trypsin can be determined with a linear range of 0.0–50.0 ng/mL with a concentration as low as 1 ng/mL. This label-free method is simple and sensitive and has been successfully used for the determination of trypsin in serum. The method can also be modified to detect other proteases. View Full-Text
Keywords: trypsin; silver nanocluster; graphene oxide; activity; Fluorimetry trypsin; silver nanocluster; graphene oxide; activity; Fluorimetry
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MDPI and ACS Style

Zhuo, C.-X.; Wang, L.-H.; Feng, J.-J.; Zhang, Y.-D. Label-Free Fluorescent Detection of Trypsin Activity Based on DNA-Stabilized Silver Nanocluster-Peptide Conjugates. Sensors 2016, 16, 1477.

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