Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A method for constructing a hydroxyapatite-based electrochemical probe and a method for measuring bace1 activity and inhibition

A technology of hydroxyapatite and chemical probes, applied in the field of biosensing, can solve the problems of low sensitivity and cumbersome operation

Inactive Publication Date: 2018-11-06
CENT SOUTH UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional method still has problems such as cumbersome operation and low sensitivity.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method for constructing a hydroxyapatite-based electrochemical probe and a method for measuring bace1 activity and inhibition
  • A method for constructing a hydroxyapatite-based electrochemical probe and a method for measuring bace1 activity and inhibition
  • A method for constructing a hydroxyapatite-based electrochemical probe and a method for measuring bace1 activity and inhibition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] A gold electrode with a diameter of 2 mm was placed in a layer containing 0.05 μm Al 2 o 3 Polish on a polishing cloth, rinse the polished electrode with secondary water, and then ultrasonically clean it with absolute ethanol and secondary water for 5 minutes to remove the polishing powder attached to the surface of the electrode, and then dry it with nitrogen. Put the treated gold electrode, Ag / AgCl reference electrode and platinum wire counter electrode together into the reaction cell containing 5mM potassium ferricyanide solution, and perform cyclic voltammetry scanning at a voltage of -0.1-0.6V. Set the scanning speed to 0.1V s -1 ,

[0066] The potential difference was 85mV, indicating that the surface of the gold electrode was cleaned. Take 5 μL of the polypeptide (CKTEEISEVNLDAEFRHDSGY) with a concentration of 4 μM and drop it on the surface of the electrode to react for 12 hours.

[0067] After the electrode is washed, take 5 μL of the peptide with a concent...

Embodiment 2

[0070] Because hydroxyapatite contains a large amount of phosphate, and alkaline phosphatase can hydrolyze sodium pyrophosphate into phosphate, and the phosphate will react with Na 2 MoO 4 The formation of phosphomolybdate precipitation in an acidic environment produces dual electrochemical signals, so the dual-signal mode was first tested. Drop 1mg / mL hydroxyapatite solution on the surface of the electrode with 6mM Na 2 MoO 4 reaction, followed by 0.5M H 2 SO 4 It is an electrolyte, within the range of 0.1 ~ 0.5V, the frequency of 15HZ sweeps the square wave voltammetry curve. Take 100μM sodium pyrophosphate solution and drop it on the surface of the electrode with Na 2 MoO 4 reaction, and reacting sodium pyrophosphate first with alkaline phosphatase and then with Na 2 MoO 4 reaction, and then likewise in 0.5M H 2 SO 4 Sweep square wave voltammetry curve in solution. Finally, the prepared electrochemical probe was mixed with Na 2 MoO 4 reaction, also at 0.5M H 2...

Embodiment 3

[0072]Put the blank gold electrode and the gold electrode modified by the peptide in the 5mM potassium ferricyanide solution reaction cell, and perform cyclic voltammetry scanning at the voltage of -0.1-0.6V, and characterize the modification of the peptide on the electrode surface by the change of the electrode peak current . Add 0 and 2.5 U / mL BACE1 solutions dropwise on the surface of the two polypeptide-modified electrodes to react for 1 hour, rinse the electrodes and then drop the hydroxyapatite probe solution on the electrode surface to react for 1 hour; then add 100 μM pyrophosphate dropwise The sodium solution was reacted for 40 minutes; finally, Na was added dropwise on the surface of the electrode 2 MoO 4 , reacted for 40 minutes, and then in 0.5M H 2 SO 4 Sweep the square wave voltammetry curve in the solution, detect it by voltammetry, and test the feasibility of the method according to the obtained corresponding peak current value.

[0073] Similarly, 0 and 2....

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses a hydroxyapatite-based electrochemical probe and a construction method thereof, and a method for determining the activity and the inhibition property of BACE1. According to the present invention, an A[beta] antibody and alkaline phosphatase are co-modified on a hydroxyapatite matrix to form the hydroxyapatite-based electrochemical probe; the preparation method comprises: sequentially placing hydroxyapatite nanoparticles in a polyethylenimine solution and carrying out a reaction, placing in a glutaraldehyde solution and carrying out a reaction, and placing in a solution containing an A[beta] antibody and alkaline phosphatase and carrying out a reaction so as to obtain the probe; and the method for determining the activity and the inhibition property of BACE1 by using the hydroxyapatite-based electrochemical probe has advantages of simpleness, rapidness, high sensitivity and wide detection range, and can be widely promoted and applied.

Description

technical field [0001] The present invention relates to an electrochemical probe, in particular to a molecular probe for measuring the activity and inhibition of BACE1, in particular to a method for constructing a hydroxyapatite-based electrochemical probe and a hydroxyapatite-based electrochemical probe. A chemical probe is used to determine the activity and inhibition of BACE1, which belongs to the technical field of biosensing. Background technique [0002] In recent years, studies on Alzheimer's disease have shown that the accumulation of β-amyloid peptide (Aβ peptide) in the brain is one of the main causes of Alzheimer's disease. Aβ peptides are produced in vivo mainly through the hydrolysis of β-like precursor protein (APP) by β-secretase (such as pre-amyloid β-decomposing enzyme, BACE1) and γ-secretase. In vivo, the activity of β-secretase plays a decisive role in the production of Aβ peptide. Inhibiting the activity of β-secretase can reduce the production of Aβ pe...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48G01N33/68G01N33/543
CPCG01N27/3277G01N27/48G01N33/5438G01N33/6896
Inventor 阳明辉
Owner CENT SOUTH UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com