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An electrochemiluminescent biosensor, its preparation method and its application in the detection of base excision repair enzymes

A luminescent bio-electrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of catalytic activity inhibition, variability, high cost of preparation, purification and storage, and achieve simplified operating procedures and sensitive detection. Effect

Active Publication Date: 2021-12-03
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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  • An electrochemiluminescent biosensor, its preparation method and its application in the detection of base excision repair enzymes
  • An electrochemiluminescent biosensor, its preparation method and its application in the detection of base excision repair enzymes
  • An electrochemiluminescent biosensor, its preparation method and its application in the detection of base excision repair enzymes

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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 electrochemiluminescent biosensor, a preparation method and its application in the detection of base excision repair enzymes. The electrochemiluminescence biosensor includes a β-CD / GO / GCE electrode and a FeMOF / AuNPs@luminol-Hairpin probe; wherein, the electrode is made by modifying graphene oxide and cyclodextrin onto a glassy carbon electrode; the The probes include: FeMOF, and AuNPs@luminol modified on FeMOF, AuNPs@luminol is also modified with a hairpin structure probe, the stem region of the hairpin structure probe is designed to be the target of the base excision repair enzyme to be tested base. The electrochemiluminescence biosensor of the present invention can triple the signal amplification to detect the base excision repair enzyme, so it has extremely high sensitivity, and can be used in biomedical research fields such as screening of base excision repair enzyme inhibitors / activators, biological sample analysis, 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 Patents(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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