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Method for preparing electrochemical immunosensor for pcsk9 protein detection

An immunosensor and protein detection technology, applied in the direction of material electrochemical variables, biological testing, material inspection products, etc., can solve the problems of long analysis time, limited sensitivity, cumbersome sample processing process, etc., achieve simple preparation process, improve sensitivity, The effect of reducing the time to capture antibodies

Inactive Publication Date: 2017-11-24
CHONGQING MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Although the traditional detection methods for PCSK9 include enzyme-linked immunosorbent assay (ELISA), fluorescent immunoassay, etc., these methods have many shortcomings, such as the sample processing process is too cumbersome, the analysis time is long, and the sensitivity is limited.

Method used

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  • Method for preparing electrochemical immunosensor for pcsk9 protein detection
  • Method for preparing electrochemical immunosensor for pcsk9 protein detection
  • Method for preparing electrochemical immunosensor for pcsk9 protein detection

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Embodiment 1

[0040] Step 1. Disperse 50 mg of nitrogen-doped multi-walled carbon nanotubes (N-MWCNTs) in 5 mL of sulfuric acid-nitric acid (H 2 SO 4 -H 3 PO 4 ) mixture, and then reacted at 140°C for 10 minutes. Then add 25 mg potassium permanganate (KMnO 4 ). The mixture was then treated at 65° C. for 10 minutes. After centrifugation and washing, vacuum-dry at 35°C;

[0041] Step 2. Use 0.3 and 0.05 μm Al respectively 2 o 3 The powder was used to polish the electrode into a mirror surface, and then the electrodes were ultrasonicated in the order of ultrapure water, absolute ethanol, and ultrapure water for 5 minutes each, and dried at room temperature for later use.

[0042] Step 3. Dispense 10 μL of 0.4mg mL -1 The electrode modification material nitrogen-doped graphene nanoribbons (N-GNRs) is dropped on the electrode surface and dried at room temperature;

[0043] Step 4. Add 8 μL n-C 60 -PdPt nanocomposite modified to the dry electrode surface, dried at room temperature;

...

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Abstract

The invention relates to a preparation method and application of an electrochemical immunosensor for a biomarker, namely a recombinant proprotein convertase subtilisin kexin type 9 (pcsk9) protein for predicting and diagnosing cardiovascular diseases, and belongs to the technical field of electrochemical detection. The preparation method is characterized by comprising the following steps: firstly, performing nitrogen doping on a graphene nanobelt (n-gnrs), performing amination on a fullerene-palladium-platinum nanoparticle (n-C60 / pdpt) composite material and a staphylococcus aureus a protein (spa), and performing layer-by-layer self-assembling so as to immobilize a pcsk9 antibody (ab1); mixing the synthesized nanoparticle-poly-methylene blue (pt-pmb) with the pcsk9 antibody (ab2), and preparing a nano beacon, thereby obtaining the electrochemical immunosensor for pcsk9 protein detection. The electrochemical immunosensor has the advantages of being high in sensitivity, good in specificity and rapid and convenient in detection. A novel method for pcsk9 protein detection is provided, and useful information is provided for clinical prediction and diagnosis on cardiovascular diseases.

Description

Technical field: [0001] The present invention relates to the preparation method and the application of the electrochemical immunosensor that can be used to predict and diagnose the biomarker of cardiovascular disease namely proprotein convertase subtilisin Kexin 9 type (PCSK9) detection, based on nitrogen-doped graphene nanobelt ( N-GNRs), aminated fullerene-palladium platinum nanoparticles (n-C 60 / PdPt) as an electrode modification material, and platinum nanoparticles-polymethylene blue (Pt-PMB) as a nano-beacon prepared sandwich-type immunosensor for detecting PCSK9, which belongs to the field of electrochemical detection. Background technique: [0002] At present, cardiovascular disease has become the main cause of human death because of its high morbidity, disability and mortality, which threaten human health and seriously affect people's quality of life. Proprotein convertase subtilisin kexin type 9 (PCSK9) is a secreted serine protease mainly synthesized by the liver...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N27/48
CPCG01N33/68G01N27/48
Inventor 于超李燕陈俊
Owner CHONGQING MEDICAL UNIVERSITY
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