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Electrochemical biosensor for detecting base excision repair enzyme and preparation method and application of electrochemical biosensor

A biosensor and electrochemical technology, applied in the field of electrochemical detection, can solve the problems of dangerous radioactive substances, time-consuming and laborious, complicated procedures, etc., and achieve the effects of low cost, simplified experimental process, and simple method.

Active Publication Date: 2020-01-14
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional DNA glycosylase detection methods include gel electrophoresis, radioactive detection, and chromatographic analysis, but these methods are usually time-consuming and laborious, and involve dangerous radioactive substances and complicated procedures

Method used

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  • Electrochemical biosensor for detecting base excision repair enzyme and preparation method and application of electrochemical biosensor
  • Electrochemical biosensor for detecting base excision repair enzyme and preparation method and application of electrochemical biosensor
  • Electrochemical biosensor for detecting base excision repair enzyme and preparation method and application of electrochemical biosensor

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preparation example Construction

[0042] In yet another specific embodiment of the present invention, a method for preparing the above-mentioned electrochemical biosensor for detecting base excision repair enzymes is provided, the preparation method comprising:

[0043] (1) Preparation of β-CD / FeCN / GCE electrode: Add FeCN solution onto the surface of GCE to obtain FeCN / GCE; after drying, add β-CD solution onto FeCN / GCE to obtain β-CD / FeCN / GCE electrode.

[0044] (2) Preparation of MB-hairpin / AuNPs probe: A gold nanoparticle (AuNP) solution and a thiolated MB-hairpin structure probe solution were mixed and incubated, centrifuged, washed and dispersed.

[0045] In yet another specific embodiment of the present invention, the concentration of the FeCN solution is 0.5 to 4 mg / mL (preferably 2 mg / ml); the concentration of the β-CD solution is 0.5 to 4 mg / mL (preferably 2 mg / ml) ; It has been verified by experiments that when the concentration of FeCN solution is 2mg / ml and the concentration of β-CD solution is 2m...

Embodiment

[0061] The principle of this embodiment, such as figure 1 Shown:

[0062] The principle of electrochemical biosensor for UDG detection is as follows: figure 1 shown. FeCN solution was dropped onto the surface of GCE to obtain FeCN / GCE. After drying at room temperature, the β-CD solution was dropped onto FeCN / GCE to form β-CD / FeCN / GCE. The DNA substrate is designed as a hairpin structure with a stem and a loop. The stem contains two complementary strands, including a methylene blue MB-labeled 3' end and a sulfhydryl-labeled 5' end with six uracil bases. The sulfhydryl group at the 5' end is connected to the AuNPs via a gold-sulfur bond, and MB cannot be recognized by β-CD / FeCN / GCE due to the steric effect of AuNPs and the hairpin stem-loop structure of DNA, resulting in a lower background current. The presence of UDG can facilitate the removal of six uracil bases from hairpin DNA, resulting in a linear single-stranded DNA with MB at the 3' end. MB can be assembled on the ...

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Abstract

The invention provides an electrochemical biosensor for detecting a base excision repair enzyme and a preparation method and application of the electrochemical biosensor. The biosensor comprises a beta-CD / FeCN / GCE electrode, the beta-CD / FeCN / GCE electrode is prepared by modifying iron rich nitrogen carbon nanotubes and cyclodextrin on a glassy carbon electrode; the electrochemical biosensor further comprises a MB-hairpin / AuNPs probe, and the MB-hairpin / AuNPs probe comprises gold nanoparticles and thiolation MB-hairpin structure probe modified on AuNP, wherein a stem area of the MB-hairpin structure probe is designed to have one or more target bases of base excision repair enzymes to be detected. The electrochemical biosensor has the advantages of high sensitivity, low background signal andthe like, and has wide application value in the fields of base excision repair enzyme inhibitor screening, biological sample analysis and the like.

Description

technical field [0001] The invention belongs to the technical field of electrochemical detection, and in particular relates to an electrochemical biosensor for detecting base excision repair enzymes, a preparation method and application thereof. Background technique [0002] The information disclosed in the Background of the Invention is only intended to increase the understanding of the general background of the invention, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes the prior art that is already known to those skilled in the art. [0003] Natural enzymes have high activity and good specificity, and play an important role in the body's metabolism and various biochemical reactions. However, natural enzymes face enormous challenges, including high costs for preparation, purification, and storage, easy denaturation under harsh conditions, and inhibition of catalytic activity in some complex media such as was...

Claims

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Application Information

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IPC IPC(8): G01N27/26G01N27/30G01N27/327
CPCG01N27/26G01N27/30G01N27/3271
Inventor 张春阳崔琳赵敏慧
Owner SHANDONG NORMAL UNIV