title: An electrochemiluminescence biosensor for endonuclease EcoRI detection.
abstract: Endonucleases cleavage of DNA plays an important role in biological and medicinal chemistry. This work was going to develop a reliable and sensitive electrochemiluminescent (ECL) biosensor for detecting endonucleases by using gold nanoparticles graphene composite (GNPs-graphene) as a signal amplifier. Firstly, the GNPs and graphene were simultaneously deposited on the glassy carbon electrode (GCE) by cyclic voltammetry. Then a stem DNA was anchored on the surface of GCE. And with a modifying DNA introduced into the electrode by DNA assembly, a strong ECL signal was obtained. After a DNA modified with ferrocene assembly to the stem DNA, the ECL signal had a sharp decrease due to the quench effect of ferrocene to and the biosensor comes into being a "off" state. With the effect of endonuclease, the ECL signal had a recovery because of the ferrocene being released and the biosensor formed a "on" state. Moreover, the recovery of ECL signal was related to the concentration of endonucleases. Combining specific defined DNA and endonuclease, this method has a potential to detect different endonucleases. In this work, we took the EcoRI as an example to identify the feasibility of ECL biosensor in applying in sensitive detection of endonucleases using a GNPs-graphene signal amplifier. Under optimal condition, the proposed biosensor obtained a low limit of detection (LOD) 5.6×10-5UmL-1. And the stability, selectivity and reproducibility of the biosensor also were researched.
authors: Department of Chemistry and Laboratory for Preparation and Application of Ordered Structural Materials of Guangdong Province, Shantou University, Shantou, Guangdong 515063, PR China.
authors: Yingjie
authors: Y
authors: Li
authors: Department of Pharmacy, Taishan Medicine College, Taian, Shandong 271016, PR China.
authors: Yuqin
authors: Y
authors: Li
authors: Department of Chemistry and Laboratory for Preparation and Application of Ordered Structural Materials of Guangdong Province, Shantou University, Shantou, Guangdong 515063, PR China.
authors: Yaoyu
authors: Y
authors: Wu
authors: Department of Chemistry and Laboratory for Preparation and Application of Ordered Structural Materials of Guangdong Province, Shantou University, Shantou, Guangdong 515063, PR China.
authors: Fushen
authors: F
authors: Lu
authors: Analysis & Testing Center, Shantou University, Shantou, Guangdong 515063, PR China.
authors: Yaowen
authors: Y
authors: Chen
authors: Department of Chemistry and Laboratory for Preparation and Application of Ordered Structural Materials of Guangdong Province, Shantou University, Shantou, Guangdong 515063, PR China; Analysis & Testing Center, Shantou University, Shantou, Guangdong 515063, PR China. Electronic address: whgao@stu.edu.cn.
authors: Wenhua
authors: W
authors: Gao
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D004247:DNA / Q000737:chemistry; D015246:Deoxyribonuclease EcoRI / Q000032:analysis*; D055664:Electrochemical Techniques / Q000379:methods*; D005296:Ferrous Compounds / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D006108:Graphite / Q000737:chemistry*; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000379:methods*; D000075163:Metallocenes; D053758:Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D015203:Reproducibility of Results
chemical_list: D005296:Ferrous Compounds; D000075163:Metallocenes; D006046:Gold; D006108:Graphite; D004247:DNA; D015246:Deoxyribonuclease EcoRI; C004998:ferrocene
keywords: Amplifier; ECL; EcoRI; Endonucleases; GNPs-graphene; Ru(bpy)(3)(2+)