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Chemical biosensor for Tau protein detection of Alzheimer's disease

A technology for Alzheimer's disease and protein detection, applied in the field of medical detection, can solve the problems of time-consuming, labor-intensive, complicated operation, etc., and achieve the effect of simple operation, low detection sensitivity, and simple and accurate detection method.

Inactive Publication Date: 2020-09-01
中玺(泉州)科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a chemical biosensor for the detection of Alzheimer's disease Tau protein, which solves the problem of measuring the content of Tau protein in CSF by conventional methods, such as enzyme-linked immunoso

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Prepare reagents and instruments: Reagents include 3-mercaptopropionic acid (MPA) (C 3 h 6 o 2 S), 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) (C 8 h 17 N 3 HCl), tris (2-carboxyethyl) phosphine hydrochloride (TCEP) (C 9 h 15 o 6 P HCl), N-hydroxysuccinimide (NHS) (C 4 h 5 NO 3 ), 2-(N-morpholino)ethanesulfonic acid monohydrate (MES) (C 6 h 13 NO 4 S), ferrocenemethanol (FcM) (C 11 h 12 FeO), L-ascorbic acid 2-phosphate sesquihydrate magnesium salt hydrate (AAP) (C 6 h 6 Mg 1.5 o 9 P xH2O), potassium ferrocyanide (K 4 Fe(CN) 6 ), potassium ferricyanide (K 3 Fe(CN) 6 ), potassium chloride (KCl), sodium chloride (NaCl), magnesium chloride (MgCl 2 ), ethanol (C 2 h 6 O), instruments include gold electrodes, electronic balances, pipette guns, ultrapure water systems, vortex mixers, super constant temperature mixers, electrochemical workstations, and other materials include Tau protein aptamers, Tau 381 Protein and its antibody (...

Embodiment 2

[0030] In embodiment one, add following operation:

[0031]In step 1, the aptamer of Tau protein was purchased from Shanghai Bioengineering Technology Co., Ltd., and the sequence of the aptamer Apt is: 5'-Biotin-GCGGAGCGTGGCAGG-3', Tau 381 The protein and its antibody (Ab) were purchased from Abcam, and bovine serum albumin (BSA) and alkaline phosphatase-labeled streptavidin (SA-ALP) were purchased from Shanghai Sangon Bioengineering Co., Ltd. The water used is ultrapure water (Milli-Q, Millipore, 18.2 MΩ.cm -1 ), all reagents were purchased and used directly without further purification, Tau 381 Protein and its antibody with 1×PBS (137 mM NaCl; 2.7 mM KCl; 8.1 mM NaCl 2 HPO 4 ; 1.8 mM KH 2 PO 4 ) to the desired concentration, Apt and SA-ALP were diluted with 1×B2 (10 mM Tris-HCl; 50 mM NaCl; 10 mM KCl; 10 mM MgCl 2 ) to the required experimental concentration, which greatly facilitates the user's production.

[0032] Prepare reagents and instruments: Reagents include 3...

Embodiment 3

[0034] In embodiment two, add following operation:

[0035] In step 2, cyclic voltammetry (CV) was carried out on CHI660A, and the continuous cyclic voltammetry was used to scan in the potential range of −0.2 to 1.6 V at a scan rate of 0.1 V / S to ensure the smooth progress of the measurement.

[0036] Prepare reagents and instruments: Reagents include 3-mercaptopropionic acid (MPA) (C 3 h 6 o 2 S), 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) (C 8 h 17 N 3 HCl), tris (2-carboxyethyl) phosphine hydrochloride (TCEP) (C 9 h 15 o 6 P HCl), N-hydroxysuccinimide (NHS) (C 4 h 5 NO 3 ), 2-(N-morpholino)ethanesulfonic acid monohydrate (MES) (C 6 h 13 NO 4 S), ferrocenemethanol (FcM) (C 11 h 12 FeO), L-ascorbic acid 2-phosphate sesquihydrate magnesium salt hydrate (AAP) (C 6 h 6 Mg 1.5 o 9 P xH2O), potassium ferrocyanide (K 4 Fe(CN) 6 ), potassium ferricyanide (K 3 Fe(CN) 6 ), potassium chloride (KCl), sodium chloride (NaCl), magnesium chloride...

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Abstract

The invention discloses a chemical biosensor for Tau protein detection of Alzheimer's disease. The method comprises the step of preparing reagents and instruments, wherein the reagent is prepared from3-mercaptopropionic acid (MPA) (C3H6O2S), 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) (C8H17N3. HCl), tris (2-carboxyethyl) phosphine hydrochloride (TCEP) (C9H15O6P.HCl), N-hydroxysuccinimide (NHS) (C4H5NO3), 2-(N-morpholino) ethane sulfonic acid monohydrate (MES) (C6H13NO4S), ferrocene methanol (FcM) (C11H12FeO) and L-ascorbic acid 2-magnesium sesquihydrate phosphate hydrate(AAP) (C6H6Mg1. 5O9P.xH2O). According to the invention, signal amplification can be realized only by adding a redox mediator and a reducing agent into an electrolyte solution; the operation is simple, and detection of an ultralow-concentration target analyte can be easily realized; and the chemical biosensor has the advantages of sensitive detection, low cost, good accuracy, interference resistance, specificity and selectivity, and provides a simple and accurate detection method for early diagnosis of Alzheimer's disease.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to a chemical biosensor for detecting Tau protein of Alzheimer's disease. Background technique [0002] The two main lesions formed in the brain by Alzheimer's disease (AD) are neurofibrillary tangles composed of tau protein and senile plaques containing amyloid-β (Aβ) protein, the most common form of dementia, in Proteins tend to misfold for several years before massive neurodegeneration. There are six subtypes of Tau protein in the human brain, and their fine homeostasis with microtubules is mainly regulated by the phosphorylation of serine and threonine in healthy neurons, which promotes the aggregation and stabilization of microtubules in the nervous system . However, under poor physiological conditions, because Tau protein has dozens of phosphorylation sites, it may undergo abnormal hyperphosphorylation at different sites, fail to stabilize microtubules, and cause n...

Claims

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Application Information

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IPC IPC(8): G01N27/48G01N27/327G01N33/68
CPCG01N27/48G01N27/3275G01N33/6896
Inventor 邱彬陶兰罗子伊傅志宏林敏
Owner 中玺(泉州)科技有限公司