Immuno-DNA binding directed template-free DNA extension and enzyme catalysis for sensitive electrochemical DNA methyltransferase activity assay and inhibitor screening

The Analyst
Ying ZhangShufeng Liu

Abstract

Sensitive and accurate determination of DNA methyltransferase (DNA Mtase) activity is highly pursued for understanding fundamental biological processes related to DNA methylation, clinical disease diagnosis and drug discovery. Herein, we propose a new electrochemical immuno-DNA sensing platform for DNA Mtase activity assay and inhibitor screening. After homogeneous DNA methylation by CpG methyltransferase (M.SssI Mtase), the methylated DNA can be specifically recruited onto an electrode via its immunological binding with the immobilized anti-5-methylcytosine antibody. The recruited methylated DNA was simultaneously used as a substrate to facilitate successive template-free DNA extension and enzyme catalysis for the dual-step signal amplification of DNA Mtase activity. The developed immuno-DNA sensing strategy effectively integrates solution-phase DNA methylation, surface affinity binding recognition, and successive template-free DNA extension and enzyme catalysis-based signal amplification, rendering a highly specific, sensitive and accurate assay of DNA Mtase activity. A low detection limit of 0.039 U mL-1 could be achieved with a high selectivity. It was also applied for efficient evaluation of various inhibitors. Current aff...Continue Reading

References

Mar 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·M FrommerC L Paul
Mar 27, 2001·Nature Reviews. Genetics·K D Robertson, A P Wolffe
Aug 11, 2001·Science·W ReikJ Walter
Feb 11, 2004·Analytical Biochemistry·Bo Yeon KimJong Seog Ahn
Oct 13, 2005·Journal of the American Chemical Society·Dominic C ChowAshutosh Chilkoti
Dec 13, 2005·Nature Clinical Practice. Oncology·Stephen B Baylin
Sep 4, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·S SamS Djebbar
Feb 9, 2010·Nucleic Acids Research·Robert J WoodPeter L Roach
Jan 19, 2011·Chembiochem : a European Journal of Chemical Biology·Renata Zofia JurkowskaAlbert Jeltsch
Aug 14, 2012·Analytical Chemistry·Ryoji Kurita, Osamu Niwa
Oct 30, 2013·Journal of the American Chemical Society·Natalie B Muren, Jacqueline K Barton
Jan 30, 2014·Analytical Chemistry·Huimin DengZhiqiang Gao
Jan 17, 2015·Nature·Dirk Schübeler
Feb 26, 2015·Chemical Communications : Chem Comm·Jiawan ZhouJinhua Chen
Feb 26, 2016·Theranostics·Wan Jun PohZhiqiang Gao
Oct 6, 2016·The Analyst·Leila SyedmoradiMichael L Norton
Jun 13, 2017·Chemical Communications : Chem Comm·Fei MaChun-Yang Zhang

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Methods Mentioned

BETA
biosensing
biosensor
biosensors
electrophoresis

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