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A biosensor for electrochemical detection of phosphorylated β-amyloid protein and its construction method

A technology of amyloid protein and biosensor, applied in the field of biosensing, can solve the problems of increased detection cost and reduced sensitivity, and achieve the effect of low detection limit, high sensitivity and simple detection process

Inactive Publication Date: 2019-06-07
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the interference of the large background of phosphate groups in actual samples, if the detection method cannot accurately identify P-Aβ, its sensitivity will be reduced, and the use of phosphorylated antibodies will greatly increase the detection cost

Method used

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  • A biosensor for electrochemical detection of phosphorylated β-amyloid protein and its construction method
  • A biosensor for electrochemical detection of phosphorylated β-amyloid protein and its construction method
  • A biosensor for electrochemical detection of phosphorylated β-amyloid protein and its construction method

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Embodiment 1

[0044] (1) Ti 4+ -TiP-Cd 2+ Preparation of Signaling Probe Nanoparticles

[0045] First, 2.5 g of docusate sodium salt (AOT) was dissolved in 16 mL of ethanol, and then 3 mL of H 3 PO 4 A cloudy solution was obtained and the precipitate was removed. Then, a mixture of tetrabutyl titanate (TBOT) and ethanol (0.875g / 12.5g) was quickly dropped into the AOT / ethanol solution, and a stable mixed solution was obtained by ultrasonic waves. The mixture was condensed and refluxed at 80° C. and stirred for 2 h. The solid product was washed several times with ethanol and deionized water and then dried to obtain TiP nanoparticles; 0.5mL TiP nanospheres (40mg / mL) were dispersed in 15mL of 10mMCd(NO 3 ) 2 React in the solution for 4-6 hours to obtain TiP-Cd 2+ nanoparticles; then TiP-Cd 2+ nanoparticles with 10mM Ti(SO 4 ) 2 The solution was reacted for 1 hour to obtain Ti with a uniform particle size and an average particle size of about 50 nm. 4+ -TiP-Cd 2+ Signaling probe nano...

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Abstract

The invention discloses a biosensor for electrochemically detecting phosphorylated beta-amyloid protein and a construction method thereof. A gold electrode is reacted with a mercaptocarboxylic acid compound for modification of carboxyl groups on the surface of the gold electrode; the carboxyl groups modified on the surface of the gold electrode are activated by an EDC / NHS mixed solution, the activated carboxyl groups and an Abeta antibody are subjected to amidation reaction, and after the amidation reaction is completed, ethanolamine is used to block unreacted sites on the surface of the goldelectrode, that is, the surface of the gold electrode is modified with the Abeta antibody; the gold electrode with the surface modified with the Abeta antibody is first subjected to antigen-antibody reaction with the phosphorylated beta-amyloid protein, and then is chelated with Ti<4+>-TiP-Cd<2+> signal probe nanoparticles to obtain the biosensor directly used for electrochemical detection. The biosensor can realize the quantitative detection of the phosphorylated beta-amyloid protein, the detection method is simple, the sensitivity is high, and the biosensor is favorable for popularization and use. .

Description

technical field [0001] The invention relates to a biosensor, in particular to a high-sensitivity electrochemical detection phosphorylated β-amyloid biosensor and a construction method thereof, belonging to the technical field of biosensing. Background technique [0002] Alzheimer's disease has become one of the major diseases threatening human health in the 21st century. It is a neurodegenerative disease characterized by progressive decline in cognitive ability, irreversible harmful impairment and observable memory impairment. . Amyloid β-protein (Aβ) is the main protein component of amyloid plaques present in the brains of Alzheimer's disease (AD) patients, and the formation of amyloid plaques is associated with the gradual accumulation of amyloid β-protein . While phosphorylation modifications are particularly abundant in amyloid β-protein aggregates, studies have shown that extracellular Aβ is phosphorylated in secreted variants of protein kinases. Phosphorylation of t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/327
Inventor 向娟尹珍珍
Owner CENT SOUTH UNIV
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