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2024年3期
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A High-Affinity Methyl-CpG-Binding Protein for Endonuclease-Free and Label-Free DNA Methyltransferase Activity Detection
A High-Affinity Methyl-CpG-Binding Protein for Endonuclease-Free and Label-Free DNA Methyltransferase Activity Detection
摘要
DNA methyltransferase(MTase)activity detection has received increasing attention as a promising biomarker and therapeutic target.However,most of these detection methods rely on endonuclease digestion and signal groups labeling.Herein,we present a novel platform for sensing DNA MTase activity that overcomes these limitations.Our approach is both endonuclease-free and label-free,utilizing a combination of a high-affinity streptavidin-methyl-CpG-binding domain(SA-MBD)protein and surface plasmon resonance(SPR)technology.The SA-MBD protein specifically recognizes a hairpin probe containing methylated CpG sites,which is treated with M.SssI MTase.This recognition event generates a corresponding SPR response signal.The limit of detection is as low as 0.016 U/mL,owing to the high-affinity of the SA-MBD protein.Notably,we have demonstrated the feasibility of our method for M.SssI MTase activity analysis in serum and inhibitor screening,which implies the potential prospects for biomedical research.
Author：
Yang Bai
Shulin Tan
Yingsong Sheng
Yueqing Gu
Haiping Wu
Baicun Li
Yunlong Liu
作者单位：
Department of Biomedical Engineering,School of Engineering,China Pharmaceutical University,Nanjing 210009,China
Department of Clinical Pharmacy,Jinling Hospital,State Key Laboratory of Analytical Chemistry for Life Science&Jiangsu Key Laboratory of Molecular Medicine,Medical School of Nanjing University,Nanjing 210002,China
School of Pharmaceutical Sciences,Southern Medical University,Guangzhou 510515,China
National Center for Respiratory Medicine,Center of Respiratory Medicine,China-Japan Friendship Hospital,Beijing 100029,China
年，卷(期)：
2024, 8(3)
机标分类号：
S825.2
R692.31
R378.91+1
在线出版日期：
2024年11月27日
