Electrochemiluminescence immunosensor for detecting beta-amyloid protein and construction of electrochemiluminescence immunosensor

A technology for amyloid and luminescent immunity, which is applied in the field of electrochemiluminescence immunosensors, can solve the problems of electrochemiluminescence immunosensors that do not see β-amyloid, and achieve ultra-sensitive detection, improve efficiency, and save reagents Effect

Active Publication Date: 2017-09-22
SHANGHAI NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there have not been any reports of electroch...

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  • Electrochemiluminescence immunosensor for detecting beta-amyloid protein and construction of electrochemiluminescence immunosensor
  • Electrochemiluminescence immunosensor for detecting beta-amyloid protein and construction of electrochemiluminescence immunosensor
  • Electrochemiluminescence immunosensor for detecting beta-amyloid protein and construction of electrochemiluminescence immunosensor

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Embodiment 1

[0062] Aβ Antibody-Nafion@Mesoporous Carbon-Ru(bpy) 3 2+ Preparation of nanocomposites

[0063] Firstly, Nafion@ mesoporous carbon-terpyridine ruthenium composite material was synthesized, and the process was as follows: 10 mg of mesoporous carbon (such as figure 1Shown in A) was dispersed in 5mL of 5wt% Nafion-ethanol mixed solvent, and stirred for 4 hours in an ice-water bath to obtain a Nafion-wrapped mesoporous carbon composite material. Nafion@ mesoporous carbon particles were obtained after centrifugation (12000rpm, 4°C), washed with PBS buffer solution, and dispersed in 2mL PBS for later use.

[0064] Under the condition of continuous stirring, 1mL 20mM ruthenium terpyridine solution was fully mixed with the above-mentioned Nafion@ mesoporous carbon composite material solution, stirred and reacted in an ice-water bath for 24 hours, centrifuged (12000rpm, 4°C) to remove excess unreacted substances, and the mesoporous carbon composite material was obtained. Pore ​​car...

Embodiment 1-1

[0067] Firstly, Nafion@mesoporous carbon-terpyridine ruthenium composites were synthesized. The process was as follows: 10mg of mesoporous carbon was dispersed in 3mL of 3wt% Nafion-ethanol mixed solvent, and stirred for 3 hours in an ice-water bath to obtain Nafion-wrapped mesoporous carbon composites. Nafion@ mesoporous carbon particles were obtained after centrifugation (12000rpm, 4°C), washed with PBS buffer solution, and dispersed in 2mL PBS for later use.

[0068] Under continuous stirring conditions, fully mix 0.5mL 25mM ruthenium terpyridine solution with the above-mentioned Nafion@ mesoporous carbon composite material solution, stir and react in an ice-water bath for 18 hours, centrifuge (12000rpm, 4°C) to remove excess unreacted substances, and obtain Mesoporous Carbon@Ru(bpy) 3 2+ The composite material was washed with PBS buffer solution, dispersed in 2mL PBS and stored in the dark at 4°C for later use.

[0069] Add 500 μL of the above mesoporous carbon@Ru(bpy) ...

Embodiment 1-2

[0071] Firstly, the Nafion@mesoporous carbon-terpyridine ruthenium composite material was synthesized. The process was as follows: 10 mg of mesoporous carbon was dispersed in 7 mL of 7wt% Nafion-ethanol mixed solvent, and stirred for 4 hours in an ice-water bath to obtain Nafion-wrapped mesoporous carbon composites. Nafion@ mesoporous carbon particles were obtained after centrifugation (12000rpm, 4°C), washed with PBS buffer solution, and dispersed in 2mL PBS for later use.

[0072] Under the condition of continuous stirring, 2mL 15mM ruthenium terpyridine solution was fully mixed with the above-mentioned Nafion@ mesoporous carbon composite material solution, stirred and reacted in an ice-water bath for 24 hours, centrifuged (12000rpm, 4°C) to remove excess unreacted substances, and the mesoporous carbon composite material was obtained. Pore ​​carbon @Ru(bpy) 3 2+ The composite material was washed with PBS buffer solution, dispersed in 2mL PBS and stored in the dark at 4°C f...

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Abstract

The invention relates to an electrochemiluminescence immunosensor for detecting beta-amyloid protein and construction of the electrochemiluminescence immunosensor. The sensor comprises a magnetic glassy carbon electrode and Fe3O4-expoxy, a mesoporous carbon @Ru(bpy)32+/beta-amyloid protein antibody composite nanometer composite material, beta-amyloid protein to be detected with different concentrations and a gold nanorod-beta- beta-amyloid protein aptamer nanometer composite material, wherein the Fe3O4-expoxy, the mesoporous carbon @Ru(bpy)32+/beta-amyloid protein antibody composite nanometer composite material, beta-amyloid protein to be detected with different concentrations and the gold nanorod-beta- beta-amyloid protein aptamer nanometer composite material are synthesized on the surface of the magnetic glassy carbon electrode in sequence. Compared with the prior art, the electrochemiluminescence immunosensor has the advantages that the constructing method is simple, detection on Alzheimer's disease markers (beta-amyloid protein and the like) is high in speed, high in sensitivity, high in specificity, low in detection limit, wide in detection range and the like, and the application of the electrochemiluminescence immunosensor provides a new thought for early diagnosis of the Alzheimer's disease and detection of markers of other diseases.

Description

technical field [0001] The invention relates to an electrochemiluminescence immunosensor, in particular to an electrochemiluminescence immunosensor for rapid and high-sensitivity detection of β-amyloid protein and its construction method and application. Background technique [0002] β-amyloid β-protein (Aβ) is hydrolyzed from β-amyloid precursor protein (APP), secreted by cells, and has a strong neurotoxic effect after the accumulation of cell matrix precipitation. And can activate a series of pathological events. Extracellular senile plaques formed by β-amyloid deposition are considered to be the main cause of degeneration and death of neurons surrounding senile plaques in the brain of Alzheimer's disease (AD) patients. AD is a brain disorder that causes the gradual loss of nerve cells in the brain. The onset and mental changes of patients are hidden, and the main manifestations are neuropsychiatric symptoms such as progressive memory impairment, personality changes, and...

Claims

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Application Information

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IPC IPC(8): G01N27/30G01N21/359G01N21/33G01N21/76G01N33/53G01N33/68
CPCG01N21/33G01N21/359G01N21/76G01N27/308G01N33/53G01N33/6896
Inventor 贾能勤柯虹王银芳沙海峰张鑫郭薇薇黄楚森
Owner SHANGHAI NORMAL UNIVERSITY
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