Electrochemical luminescent biosensor, preparation method and application of electrochemical luminescent biosensor to detection of base excision repair enzyme

A luminescent biological and electrochemical technology, applied in the fields of electrochemical variables of materials, scientific instruments, instruments, etc., can solve the problems of high preparation, purification and storage costs, inhibition of catalytic activity, and variability, and achieve a wide range of potential applications and high sensitivity. , the effect of simplifying the operation procedure

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

However, peroxidases are costly to prepare, purify, and store, are volatile under harsh conditions, and their catalytic activity is inhibited in some complex media (such as wastewater)

Method used

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  • Electrochemical luminescent biosensor, preparation method and application of electrochemical luminescent biosensor to detection of base excision repair enzyme
  • Electrochemical luminescent biosensor, preparation method and application of electrochemical luminescent biosensor to detection of base excision repair enzyme
  • Electrochemical luminescent biosensor, preparation method and application of electrochemical luminescent biosensor to detection of base excision repair enzyme

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Experimental program
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preparation example Construction

[0047] In yet another specific embodiment of the present invention, a method for preparing the above-mentioned electrochemiluminescent biosensor for detecting base excision repair enzymes is provided, the preparation method comprising:

[0048] (1) Preparation of β-CD / GO / GCE electrode: GO solution was added dropwise on the surface of GCE to obtain GO / GCE; after drying, β-CD solution was added dropwise onto GO / GCE to obtain β-CD / GO / GCE electrodes;

[0049] (2) Preparation of FeMOF / AuNPs@luminol-Hairpin probe: Mix FeMOF and AuNPs@luminol to obtain FeMOF / AuNPs@luminol, then mix and incubate FeMOF / AuNPs@luminol with hairpin probe solution, and centrifuge to obtain .

[0050] In yet another specific embodiment of the present invention, the concentration of the β-CD solution is 1 to 5 mM (preferably 2 mM); it has been verified by experiments that when the concentration of the β-CD solution is 2 mM, the luminous intensity is the highest and the detection effect is the best. good. ...

Embodiment

[0072] The principle of this embodiment, such as figure 1 Shown:

[0073] A hairpin probe with 6 uracil bases in the stem, a sulfhydryl group at the 5' end was designed for immobilization on the surface of FeMOF / AuNPs@luminol, and an Fc at the 3' end. In the absence of UDG, the six uracil bases of the hairpin probe could not be deleted, and the FeMOF / AuNPs@luminol-hairpin probe could not be captured to the electrode surface by β-CD, resulting in no Prussian blue production. Due to the absence of Fc and Prussian blue on the electrodes, the ECL luminescence intensity is lower. When UDG exists, 6 uracil bases in the hairpin DNA are removed, and 6 AP sites are generated, forming a straight single-stranded DNA (ssDNA) with Fc at the 3' end. Fc is immobilized on the electrode through the host-guest interaction with β-CD to further catalyze luminol free radicals (L -· ) and H 2 o 2 generated OH · The reaction produces 3-carbamate (AP 2- *). When AP 2- *When returning to the ...

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Abstract

The invention provides an electrochemical luminescent biosensor, a preparation method and application of the electrochemical luminescent biosensor to detection of a base excision repair enzyme. The electrochemical luminescent biosensor comprises a beta-CD/GO/GCE electrode and a FeMOF/AuNPs@luminol-Hairpin probe, wherein the electrode is prepared through modification of graphene oxide and cyclodextrin to a glassy carbon electrode; the probe comprises FeMOF and AuNPs@luminol modified on the FeMOF; a hairpin structure probe is also modified on the AuNPs@luminol; and a target base of the base excision repair enzyme to be detected is designed in a stem area of the hairpin structure probe. The electrochemical luminescent biosensor provided by the invention can be used for detecting the base excision repair enzyme through triple signal amplification, so that the electrochemical luminescent biosensor has extremely high sensitivity, and can be applied in the biomedical research fields of screening of base excision repair enzyme inhibitors/activators, analysis of biological samples and the like.

Description

technical field [0001] The invention belongs to the technical field of electrochemiluminescence detection, and in particular relates to an electrochemiluminescence biosensor, a preparation method and its application in the detection of base excision repair enzymes. 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] Metal-organic frameworks (MOFs) consist of metal ions or clusters linked by organic linkers, and possess superior physical and chemical properties. MOFs have the characteristics of large pore volume, large surface area, multiple topological structures, adjustable pore size, and good surface chemical properties, and have broad application prospects in...

Claims

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Application Information

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