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Sensors 2016, 16(11), 1477; doi:10.3390/s16111477

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

1,2,†
,
1,2,†
,
1,2
and
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
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editors: Jong Seung Kim and Min Hee Lee
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)
View Full-Text   |   Download PDF [1620 KB, uploaded 9 November 2016]   |  

Abstract

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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