Preparation method of renewable electrochemical immunosensor for sCD40L detection

An immunosensor, electrochemical technology, applied in the direction of electrochemical variables of materials, scientific instruments, instruments, etc., can solve the problems of uneven dispersion and easy agglomeration, and achieve the effects of firm bonding, enhanced conductivity, and fast detection speed.

Inactive Publication Date: 2017-03-08
CHONGQING MEDICAL UNIVERSITY
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  • Abstract
  • Description
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Problems solved by technology

However, due to the existence of π-π electrons, it forms the interaction of van der Waals force and has strong hydrophobicity, which leads to its uneven dispersion and easy aggregation in many solvents, so its application in electrochemical immunosensors has been subject to many limit

Method used

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  • Preparation method of renewable electrochemical immunosensor for sCD40L detection
  • Preparation method of renewable electrochemical immunosensor for sCD40L detection
  • Preparation method of renewable electrochemical immunosensor for sCD40L detection

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Embodiment 1

[0035] Step 1. Weigh 2mg of c-MWCNTs into 2mL of ultrapure water and sonicate for 1-3h to disperse evenly. Add an appropriate amount of EDC and NHS solution to activate its carboxyl group under constant stirring, and then add 100 μL PEI solution to continue stirring for 2-5 hours after magnetic stirring for 30 minutes. After the above materials were washed several times, they were dispersed in 2 mL of ultrapure water, and 100 μL of HAuCl were added successively. 4 ·6H 2 O (1%) and 900 μL of NaBH 4 (30mM) stirred overnight, washed several times and dispersed in 2mL ultrapure water to obtain the c-MWCNTs-PEI-AuNPs nanocomposite, which was dispersed in 2mL ultrapure water and stored in a 4°C refrigerator for later use;

[0036] Step 2. Use 0.3μm and 0.05μm Al respectively 2 o 3 Polish the electrode to the mirror surface with powder, then use appropriate amount of ultrapure water, absolute ethanol, and ultrapure water to ultrasonically clean each electrode for 5 minutes in the...

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Abstract

The invention relates to a preparation method of a renewable electrochemical immunosensor capable of being used for detecting a biomarker for forecasting and diagnosing an acute coronary syndrome, i.e., a soluble CD40 ligand (sCD40L), and belongs to the technical field of electrochemical detection. The preparation method is characterized by comprising the following step: immobilizing an sCD40L antibody through a carboxyl functionalized multi-wall carbon nano tube-polyethyleneimine-gold nanoparticle nano compound (c-MWCNTs-PEI-AuNPs) serving as a substrate material to realize capturing of sCD40L and further quantitatively detecting the sCD40L. A c-MWCNTs-PEI-AuNps nano compound is easy to prepare, high in conductivity, relatively high in stability and relatively large in specific surface area, so that a large quantity of antibodies can be firmly immobilized; furthermore, by the use of specificity identification of the antibodies and antigens, the built electrochemical immunosensor is relatively high in specificity. The electrochemical immunosensor has the advantages of high sensitivity, high specificity, convenience, quickness and capability of being repetitively used for multiple times; a new method is supplied to detection of the sCD40L, and available information is supplied to clinical forecasting and diagnosis of the acute coronary syndrome.

Description

[0001] Technical field: [0002] The present invention relates to a method for the preparation of an electrochemical immunosensor for the detection of soluble CD40 ligand (sCD40L), which can be used as a good and reliable biochemical marker for prediction and diagnosis of acute coronary syndrome, based on carboxyl-functionalized multi-walled carbon Direct regenerative immunosensors of nanotubes (c-MWCNTs), polyethyleneimine (PEI), and gold nanoparticles (AuNPs) nanocomposites for the detection of sCD40L belong to the field of electrochemical detection. [0003] Background technique: [0004] At present, cardiovascular disease has become a major cause of human death due to its high morbidity, disability and mortality, threatening human health, and acute coronary syndrome is a common and serious cardiovascular disease in clinical practice. . sCD40L is a member of the tumor necrosis factor superfamily and belongs to type I transmembrane glycoprotein, which can promote cell expans...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/543G01N27/48
CPCG01N33/5302G01N27/48G01N33/54346G01N2800/324
Inventor 于超吴静赵一璘牛亚珍
Owner CHONGQING MEDICAL UNIVERSITY
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