Electrochemical synthesis-based acetyl transferase electrochemical light-emitting sensor built by Ru-MOF and application of acetyl transferase electrochemical light-emitting sensor

An acetyltransferase and electrochemical technology, which is applied in the field of functional biomaterials and biosensing, can solve the problems of undisclosed acetyltransferase electrochemiluminescence sensors, etc., and achieve the effects of good selectivity, high sensitivity, and low dosage

Active Publication Date: 2020-01-10
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no reports on the construction of acetyltransferase ECL sensors and their applications based on electrochemically synthesized Ru-MOF at home and abroad.

Method used

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  • Electrochemical synthesis-based acetyl transferase electrochemical light-emitting sensor built by Ru-MOF and application of acetyl transferase electrochemical light-emitting sensor
  • Electrochemical synthesis-based acetyl transferase electrochemical light-emitting sensor built by Ru-MOF and application of acetyl transferase electrochemical light-emitting sensor
  • Electrochemical synthesis-based acetyl transferase electrochemical light-emitting sensor built by Ru-MOF and application of acetyl transferase electrochemical light-emitting sensor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 Preparation of sensor

[0035] The technical scheme adopted by the present invention to solve the above-mentioned technical problems is: construct an acetyltransferase electrochemiluminescent sensor and its application based on electrochemically synthesized Ru-MOF, and the specific steps are as follows:

[0036] Preparation of Electrode 1:

[0037] First, the glassy carbon electrode (GCE, 3 mm in diameter) was polished on the suede with Al2O3 powder (0.05 μm) for 5 min. N 2 Blow dry to get a bare glassy carbon electrode. Labeled Electrode 1.

[0038] Preparation of Electrode 2:

[0039] Liquid A: ZnCl 2 (0.9mmol, 0.123g), KNO 3 (0.1M, 0.0303g), dissolved in 3mL of water. Solution B: Dissolve trimesic acid (0.5 mmol, 0.105 g) in 3 mL of ethanol.

[0040] Solution A and solution B are mixed at room temperature, 5mmol of Ru(bpy) 3 2+ Add in the mixture of A and B, under vigorous stirring, stir at room temperature for 3h. The solution is ready for us...

Embodiment 2

[0045] Embodiment 2 Feasibility experiment

[0046] The technical solution adopted by the present invention to solve the above technical problems is: a kind of electrochemically synthesized Ru-MOF based on electrochemically synthesized acetyltransferase electrochemiluminescence sensor and its application, the specific steps are as follows:

[0047] (1) Electrode 1-Electrode 4 were prepared as in Example 1, and their electrochemiluminescence responses in PBS (0.1 M, pH 7.0, containing 0.1 M KCl and 100 mM triethanolamine) solution were detected. Using the BPCL T15 measurement and analysis system, set the high voltage to 800V, the scan rate to 100mV / s, and the scan voltage to 0-1.35V. from figure 1 It can be seen that Electrode 2 has an obvious electrochemical response signal in PBS (0.1M, pH 7.0, containing 0.1M KCl and 100mM triethanolamine), which proves that Ru-MOF was successfully synthesized. Electrode 4 also has an obvious electrochemiluminescence response signal, and t...

Embodiment 3

[0049] Example 3 Detection of p300 activity

[0050] In the preparation process of Example 1 Electrode 4, the concentration of p300 was changed, and the final concentration of p300 was controlled to be: 0, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 50, 100, 200, 500, 800, 1000nM, such as Example 1 Prepare a series of electrochemiluminescent sensors. Experimental results such as image 3 As shown, the Cu(II)-quenched ECL signal recovered more with increasing p300 concentration. The linear correlation equation of the luminous intensity of the sensor to the logarithmic value of the p300 concentration is y=2.49x+5.43, R 2 =0.9970, the linear range is 0.01-500nM, and the detection limit is 0.001nM, indicating that the sensor can realize wide-range and high-sensitivity detection of p300 activity.

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Abstract

The invention discloses an electrochemical synthesis-based acetyl transferase electrochemical light-emitting sensor built by Ru-MOF and application of the acetyl transferase electrochemical light-emitting sensor. Ru(bpy) <3><2+>- functional MOF thin film is synthesized by an electrochemical auxiliary self-assembly method, the acetyl transferase electrochemical light-emitting sensor is simple synthesis step and relatively short in time and is friendly to environment. An ECL signal of the Ru-MOF can be quenched by Cu (II), an acetylation liquid is added, an acetyl product CoA and Cu (II) react to form a coper-sulfydryl coordination polymer, and the ECL signal is restored. On the basis, an electrochemical light-emitting biosensor is built and is applied to detection of histone acetyl transferase. The electrochemical light-emitting sensor provided by the invention has the advantages of performance stability and high sensitivity in application and can be widely applied to the field of clinic and medicine.

Description

technical field [0001] The invention relates to an electrochemiluminescence sensor and a detection method thereof, in particular to the construction of an acetyltransferase electrochemiluminescence sensor based on electrochemically synthesized Ru-MOF and its application, and belongs to the technical field of functional biomaterials and biosensing. Background technique [0002] Epigenetics refers to heritable gene expression changes other than DNA sequence changes. This kind of gene expression regulation that affects gene transcription activity without involving DNA sequence changes is called epigenetic transcriptional regulation. Histone acetylation is a gene expression modification. An important mechanism of transcriptional regulation. Chromatin structure remodeling caused by histone post-translational modifications plays an important role in the regulation of eukaryotic gene expression. Histone acetylation is modified by the interaction of two enzymes, histone acetyltrans...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N27/327
CPCG01N21/76G01N27/327
Inventor 胡宇芳胡丹丹詹甜玉郑宇迪刘鑫达冉平建王邃郭智勇
Owner NINGBO UNIV
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