Electrochemiluminescent biosensor for detection of endonuclease, preparation and application thereof

A luminescent bio-electrochemical technology, applied in the direction of chemiluminescence/bioluminescence, electrochemical variables of materials, and analysis by making materials undergo chemical reactions, can solve the problems of high price, complicated detection equipment and processing process, and achieve high Specificity, rapid and accurate detection, and good stability

Active Publication Date: 2016-06-15
SHANTOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide an electrochemiluminescent biosensor for detecting endonuclease and its pre...

Method used

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  • Electrochemiluminescent biosensor for detection of endonuclease, preparation and application thereof
  • Electrochemiluminescent biosensor for detection of endonuclease, preparation and application thereof
  • Electrochemiluminescent biosensor for detection of endonuclease, preparation and application thereof

Examples

Experimental program
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Embodiment 1

[0047] A method for preparing an electrochemiluminescent biosensor for detecting endonuclease EcoRI, comprising the following steps:

[0048] (1) Glassy carbon electrode pretreatment: Glassy carbon electrode is treated with 0.05μm α-A1 2 o 3 After the polishing powder is polished, it is rinsed with deionized water, and ultrasonicated in deionized water and ethanol for 10 min respectively. Using a three-electrode system for detection, the working electrode is a glassy carbon electrode, the counter electrode is a platinum wire electrode, and the reference electrode is an Ag / AgCl electrode. In 0.5M sulfuric acid, set the voltage at -0.2 to 1.6V to cycle the glassy carbon electrode. Voltammetric scanning, after the detection, rinse the electrode with deionized water, dry the surface of the electrode, and set aside.

[0049] (2) Preparation of gold nano-graphene modified glassy carbon electrode: at 0.5mgmL -1 In the mixture of graphite oxide (GO) and 20mM chloroauric acid, cycli...

Embodiment 2

[0067] A method for preparing an electrochemiluminescent biosensor for detecting endonuclease EcoRI, comprising the following steps:

[0068] (1) Glassy carbon electrode pretreatment: Glassy carbon electrode is treated with 0.3μm α-A1 2 o 3 After the polishing powder is polished, it is rinsed with deionized water, and ultrasonicated in deionized water and ethanol for 10 min respectively. Using a three-electrode system for detection, the working electrode is a glassy carbon electrode, the counter electrode is a platinum wire electrode, and the reference electrode is an Ag / AgCl electrode. In 0.5M sulfuric acid, set the voltage at -0.2 to 1.6V to cycle the glassy carbon electrode. Voltammetric scanning, after the detection, rinse the electrode with deionized water, dry the surface of the electrode, and set aside.

[0069] (2) Preparation of gold nano-graphene modified glassy carbon electrode: at 1mgmL -1 In the mixture of graphite oxide and 60mM chloroauric acid, cyclic voltam...

Embodiment 3

[0083] A method for preparing an electrochemiluminescent biosensor for detecting endonuclease ApaI, comprising the following steps:

[0084] (1) Glassy carbon electrode pretreatment: Glassy carbon electrode is treated with 0.3μm α-A1 2 o 3 After the polishing powder is polished, it is rinsed with deionized water, and ultrasonicated in deionized water and ethanol for 10 min respectively. Using a three-electrode system for detection, the working electrode is a glassy carbon electrode, the counter electrode is a platinum wire electrode, and the reference electrode is an Ag / AgCl electrode. In 0.5M sulfuric acid, set the voltage at -0.2 to 1.6V to cycle the glassy carbon electrode. Voltammetric scanning, after the detection, rinse the electrode with deionized water, dry the surface of the electrode, and set aside.

[0085] (2) Preparation of gold nano-graphene modified glassy carbon electrode: at 1mgmL -1 In the mixture of graphite oxide and 60mM chloroauric acid, cyclic voltamm...

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Abstract

The invention relates to an electrochemiluminescent biosensor for detection of endonuclease, preparation and application thereof. The preparation steps include: S01. pretreating a glassy carbon electrode; S02. preparing a gold nano-graphene modified glassy carbon electrode by means of cyclic voltammetry; S03. respectively preparing an Ru(bpy)3<2+> labeled B-DNA beacon with a concentration of 1-10microM and a ferrocene labeled C-DNA beacon with a concentration of 1-10microM; S04. immersing the gold nano-graphene modified glassy carbon electrode obtained in S02 into mercapto modified A-DNA to conduct cultivation for 3-6h, and then conducting immersion in the Ru(bpy)3<2+> labeled B-DNA beacon and the ferrocene labeled C-DNA beacon obtained in S03 in order for 20-40min. The electrochemiluminescent biosensor provided by the invention can be used for detecting endonucleases EcoRI and ApaI. The sensor provided by the invention has good stability and reproducibility and high specificity to endonucleases EcoRI and ApaI, is hardly interfered by other substances in a detection sample, has very strong sensitivity, and is simple, convenient and quick to operate during detection, thus having certain help to research and development of antibacterial and antiviral drugs.

Description

technical field [0001] The invention relates to the field of analytical chemistry and electrochemiluminescence sensor analysis, in particular to an electrochemiluminescence biosensor for detecting endonucleases based on a gold nanometer-graphene composite material and a preparation method and application thereof. Background technique [0002] Endonucleases are a class of nucleic acids that mainly exist in prokaryotes. It is known as molecular scissors and is used to cut phosphate bonds at specific locations. Endonucleases have been widely used in PCR experiments, gene profiling, medicinal chemistry, catalytic amplification technology, etc. In prokaryotes, endonucleases protect the internal genes of the organism against foreign genes. Therefore, endonuclease is considered to be an important target substance in the field of discovery of new antibacterial and antiviral drugs. [0003] The current research methods for the detection of endonucleases include high performance li...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N27/48G01N27/30
CPCG01N21/76G01N27/30G01N27/48
Inventor 高文华李英杰陈耀文鲁福身张歆陈汉佳
Owner SHANTOU UNIV
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