Renewable electrochemical immunosensor preparation method for sCD40L detection

An immunosensor, electrochemical technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., can solve the problems of uneven dispersion, easy agglomeration, etc., to achieve the effect of strong bonding, enhanced conductivity, and promotion of development

Pending Publication Date: 2019-06-25
CHONGQING MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
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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 biosensors has been widely accepted. limit

Method used

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  • Renewable electrochemical immunosensor preparation method for sCD40L detection
  • Renewable electrochemical immunosensor preparation method for sCD40L detection
  • Renewable electrochemical immunosensor preparation method for sCD40L detection

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Embodiment 1

[0029] 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;

[0030] 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 ultrasonicate the electrode for 5 minutes in the above o...

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Abstract

The invention relates to a preparation method of a renewable electrochemical immunosensor which can be used for predicting and diagnosing a biomarker of acute coronary syndrome, namely a soluble CD40ligand (sCD40L), which belongs to the technical field of electrochemical detection. The method provided by the invention comprises the steps that a carboxyl-functionalized multi-walled carbon nanotube-polyethyleneimine-gold nanoparticle nanocomposite (c-MWCNTs-PEI-AuNPs) is used as a substrate material for immobilizing an sCD40L antibody, so as to capture the sCD40L; and then the sCD40L is quantitatively detected. Since the c-MWCNTs-PEI-AuNPs nanocomposite has the advantages of simple preparation, good electrical conductivity, good stability and large specific surface area, the nanocomposite can firmly immobilize a large number of antibodies and utilizes specific recognition of antibodies and antigens. The constructed electrochemical immunosensor has strong specificity. The method providedby the invention has the advantages of high sensitivity and specificity, convenience, quickness and regenerability, provides a new method for detecting the sCD40L, and provides useful information forclinical prediction and diagnosis of acute coronary syndrome.

Description

Technical field: [0001] The present invention relates to a preparation method and application 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. Direct regenerative immunosensors based on composites of walled carbon nanotubes (c-MWCNTs), polyethyleneimine (PEI), and gold nanoparticles (AuNPs) for the detection of sCD40L belong to the field of electrochemical detection. Background technique: [0002] 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 expansion and migration. Studies have shown th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48G01N27/327
Inventor 于超周媛吴静
Owner CHONGQING MEDICAL UNIVERSITY
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