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Electrochemical luminescence Faraday cage immunosensor for detecting histone acetyltransferase

A technology of immunosensor and acetyltransferase, which is applied in the field of functional materials and biosensing to achieve the effects of improving detection sensitivity, enhancing luminescence signal, and high specificity

Active Publication Date: 2018-09-21
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no related reports on the construction of electrochemiluminescent Faraday cage immunosensor based on DNA nanotechnology and its use in the detection of histone acetyltransferase p300 at home and abroad.

Method used

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  • Electrochemical luminescence Faraday cage immunosensor for detecting histone acetyltransferase
  • Electrochemical luminescence Faraday cage immunosensor for detecting histone acetyltransferase
  • Electrochemical luminescence Faraday cage immunosensor for detecting histone acetyltransferase

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specific Embodiment 1

[0043] An electrochemiluminescent Faraday cage immunosensor for detecting histone acetyltransferase, the specific steps are as follows:

[0044] (1) Preparation of capture unit

[0045] a. Iron ion mixed solution preparation

[0046] 0.8g FeCl 3 ·6H 2 O and 0.3g FeCl 2 4H 2 O was dissolved in 50mL of secondary water and mixed well. Ready to use.

[0047] b. Magnetic graphene oxide (nanoFe 3 o 4 Synthesis of @GO)

[0048] Take 40 mg of graphene oxide (GO) in 40 mL of secondary water, after ultrasonication for 30 min, add 50 mL of the above iron ion mixed solution, stir rapidly, raise the temperature of the above mixed solution to 85 ° C, and add dropwise under rapid stirring and reflux 25% ammonia water (NH 4 OH) until the pH of the solution was 10, and reacted for 45 minutes, the color of the solution changed from brown to black, and after cooling to room temperature, the magnetism of the obtained mixture was used, and the secondary water was repeatedly washed until ...

specific Embodiment 2

[0064] An electrochemiluminescent Faraday cage immunosensor for detecting histone acetyltransferase, the specific steps are as follows:

[0065] (1) Preparation of capture unit

[0066] a. Iron ion mixed solution preparation

[0067] 0.4g FeCl 3 ·6H 2 O and 0.15g FeCl 2 4H 2 O was dissolved in 25mL of secondary water and mixed well. Ready to use.

[0068] b. Magnetic graphene oxide (nanoFe 3 o 4 Synthesis of @GO)

[0069] Take 30mg of graphene oxide (GO) in 30mL of secondary water, after ultrasonication for 40min, add 38mL of the above iron ion mixed solution, stir rapidly, raise the temperature of the above mixed solution to 85°C, and add 28 % ammonia water (NH 4 OH) until the pH of the solution was 10, and reacted for 40 minutes, the color of the solution changed from brown to black. After cooling to room temperature, the magnetic properties of the obtained mixture were used, and the secondary water was repeatedly washed until the pH of the supernatant was neutral,...

specific Embodiment 3

[0085] An electrochemiluminescent Faraday cage immunosensor for detecting histone acetyltransferase, the specific steps are as follows:

[0086] (1) Preparation of capture unit

[0087] a. Iron ion mixed solution preparation

[0088] 1.2g FeCl 3 ·6H 2 O and 0.45g FeCl 2 4H 2 O was dissolved in 75mL of secondary water and mixed well. Ready to use.

[0089] b. Magnetic graphene oxide (nanoFe 3 o 4 Synthesis of @GO)

[0090] Take 20 mg of graphene oxide (GO) in 20 mL of secondary water, after ultrasonication for 35 min, add 25 mL of the above iron ion mixed solution, stir rapidly, raise the temperature of the above mixed solution to 85 ° C, and add 20 % ammonia water (NH 4 OH) until the pH of the solution was 10, and reacted for 50 minutes, the color of the solution changed from brown to black. After cooling to room temperature, the magnetic properties of the obtained mixture were used, and the secondary water was repeatedly washed until the pH of the supernatant was neut...

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Abstract

The invention discloses an electrochemical luminescence Faraday cage immunosensor for detecting histone acetyltransferase. The electrochemical luminescence Faraday cage immunosensor is characterized by comprising the following steps: (1) preparing a capturing unit: synthesizing magnetic graphene oxide (GO); treating the magnetic graphene oxide with an EDC / NHS (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride / N-Hydroxysuccinimide) solution and HCl in sequence; then combining with a capturing antibody Ab1; incubating at room temperature; finally adding BSA (Bovine Serum Albumin) to seal a non-specific binding site on the surface of the GO to obtain the capturing unit Ab1-nanoFe3O4@GO; (2) preparing a signal unit: connecting a recognition antibody Ab2 into a DNA (Deoxyribonucleic Acid) tetrahedron to synthesize tetrahedron-Ab2; then synthesizing nano-gold (AuNPs) and forming a compound GO@AuNPs by the nano-gold and graphene oxide; then combining the compound GO@AuNPs with tetrahedron-Ab2; then adding a Ru luminous body to prepare the signal unit; (3) preparing the Faraday cage immunosensor: dropping and coating an Ab1-nanoFe3O4@GO solution to the surface of an MGCE (Modified Glassy Carbon Electrode) to obtain Ab1-nanoFe3O4@GO / MGCE and recording as an electrode a; taking a p300 solution and dropping and coating on the Ab1-nanoFe3O4@GO / MGCE to obtain p300 / Ab1-nanoFe3O4@GO / MGCE, and recording as an electrode b; finally, taking a Ru-GO@AuNPs-tetrahedron-Ab2 solution and dropping and coating on the p300 / Ab1-nanoFe3O4@GO / MGCE to obtain a Ru-GO@AuNPs-tetrahedron-Ab2 / p300 / Ab1-nanoFe3O4@GO / MGCE, and recording as an electrode c, i.e., the electrochemical luminescence Faraday cage immunosensor.

Description

technical field [0001] The invention relates to a method for detecting histone acetyltransferase, in particular to a method for constructing an electrochemiluminescence Faraday cage immunosensor for detecting histone acetyltransferase, and belongs to the technical field of functional materials and biosensing. Background technique [0002] Histone acetyltransferase (HAT) is a biological enzyme that regulates the initiation of transcription by reversibly modifying lysine residues at the tail of core histones, thereby mediating gene activation and repression. Histone acetyltransferases and their complexes participate in many important physiological processes related to gene transcription regulation in organisms, such as: DNA replication, repair, chromosome assembly and cell cycle regulation. In addition, the disorder of histone acetylation or the abnormal function of acetyltransferase is related to the formation of tumors, especially the formation of leukemia. Metabolic syndrom...

Claims

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Application Information

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IPC IPC(8): G01N33/535
CPCG01N33/535
Inventor 胡宇芳张青青徐利华王娇饶家佳郭智勇王邃
Owner NINGBO UNIV
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