A Label-Free Fluorescence Aptasensor Based on G-Quadruplex/Thioflavin T Complex for the Detection of Trypsin

A novel, label-free fluorescent assay has been developed for the detection of trypsin by using thioflavin T as a fluorescent probe. A specific DNA aptamer can be combined by adding cytochrome c. Trypsin hydrolyzes the cytochrome c into small peptide fragments, exposing the G-quadruplex part of DNA a...

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Bibliographic Details
Published in:Molecules (Basel, Switzerland) Switzerland), 2022-09, Vol.27 (18), p.6093
Main Authors: Gu, Pan, Lu, Yangfan, Li, Shanni, Ma, Changbei
Format: Article
Language:English
Subjects:
Antibodies
Aptamers
Biological properties
Biological samples
Biosensors
Cytochrome c
Deoxyribonucleic acid
Detectors
DNA
DNA aptamer
Enzymes
Fluorescence
Fluorescent indicators
label-free
Methods
Pancreatitis
Peptides
Proteins
Quantum dots
thioflavin T
Trypsin
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Summary:A novel, label-free fluorescent assay has been developed for the detection of trypsin by using thioflavin T as a fluorescent probe. A specific DNA aptamer can be combined by adding cytochrome c. Trypsin hydrolyzes the cytochrome c into small peptide fragments, exposing the G-quadruplex part of DNA aptamer, which has a high affinity for thioflavin T, which then enhances the fluorescence intensity. In the absence of trypsin, the fluorescence intensity was inhibited as the combination of cytochrome c and the DNA aptamer impeded thioflavin T’s binding. Thus, the fluorescent biosensor showed a linear relationship from 0.2 to 60 μg/mL with a detection limit of 0.2 μg/mL. Furthermore, the proposed method was also successfully employed for determining trypsin in biological samples. This method is simple, rapid, cheap, and selective and possesses great potential for the detection of trypsin in bioanalytical and biological samples and medical diagnoses.
ISSN:1420-3049
1420-3049
DOI:10.3390/molecules27186093