Homogeneous Electrochemical Assay for Real-time Monitoring of Exonuclease III Activity Based on a Graphene Monolayer
- Authors
- Lee, Heon-Kyu; Heo, Jihye; Myung, Sung; Shin, Ik-Soo; Kim, Tae Hyun
- Issue Date
- Jul-2017
- Publisher
- Wiley - V C H Verlag GmbbH & Co.
- Keywords
- Electrochemical assay; exonuclease III activity assay; Exo III; graphene monolayer; real-time; drug screening; inhibition assay
- Citation
- Electroanalysis, v.29, no.7, pp 1749 - 1754
- Pages
- 6
- Journal Title
- Electroanalysis
- Volume
- 29
- Number
- 7
- Start Page
- 1749
- End Page
- 1754
- URI
- https://scholarworks.bwise.kr/sch/handle/2021.sw.sch/7433
- DOI
- 10.1002/elan.201700006
- ISSN
- 1040-0397
1521-4109
- Abstract
- A homogeneous electrochemical assay based on a graphene monolayer electrode was developed for simple, sensitive, rapid and quantitative analysis of the exonuclease III (Exo III) activity. The method utilized a methylene blue (MB) tagged DNA substrate with hairpin structure, and a graphene monolayer attached on the working electrode. Before digestion, the hairpin structure prevents the adsorption of the DNA substrate to the graphene surface. Degradation of the substrate by the 3'-5' Exo III, however, yields single-stranded DNA (ssDNA), resulting in its subsequent binding to the graphene surface through pi-pi stacking, which produces the voltammetric current from electrochemical reduction of the MB tag anchoring at the end of ssDNA. A direct quantification of the Exo III activity can be achieved by measuring the reductive peak current of the MB tag under easily attainable potential (similar to -0.1V vs Ag/AgCl) range comparably sensitive to the conventional methods such as a gel-based or fluorescence-based assays. Our approach can be applied to measure various exonucleases activity by adjusting the structure of DNA substrate suggesting a new assay method in drug screening and basic research related to the enzymes.
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Collections - College of Natural Sciences > Department of Chemistry > 1. Journal Articles
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