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Electronic mediator type enzymatic electrochemical immunodetection method of p53 protein

A detection method and electrochemical technology, applied in the direction of material electrochemical variables, measuring devices, scientific instruments, etc., to achieve the effects of wide linear range, low detection limit, and high sensitivity

Inactive Publication Date: 2015-04-01
SOUTHEAST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The effective combination of nanofibers and enzymatic electron mediators for the construction of electron mediator-type enzymatic electrochemical immunosensors has not been reported at home and abroad. The application of its specificity and high sensitivity to detect tumor protein p53 is an important topic worthy of research. topic

Method used

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  • Electronic mediator type enzymatic electrochemical immunodetection method of p53 protein
  • Electronic mediator type enzymatic electrochemical immunodetection method of p53 protein
  • Electronic mediator type enzymatic electrochemical immunodetection method of p53 protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0057] (1) Preparation of immune molecule immobilized active interface:

[0058] 3.2 g of nylon 6 (PA6, viscosity 2.2) and 5 mg of carboxylated multi-walled carbon nanotubes (MWCNTs) were dissolved in 10 mL of mixed solvent (m-cresol:formic acid=6:4, v / v), and the mixture Stirring was continued at room temperature for 12 hours to obtain a homogeneous and transparent precursor electrospinning solution.

[0059] During electrospinning, the uniform and transparent precursor electrospinning solution prepared above is placed in a syringe, the positive electrode of the high-voltage electrostatic generator is connected to the needle of the syringe, the negative electrode is connected to a clean bare electrode, and a micro-syringe pump is used to supply the liquid. The high-voltage static electricity generated by the high-voltage electrostatic generator is directly applied to the needle of the syringe, and the composite nanofibers (PA6-MWNTs) are collected on the bare electrode;

[0...

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Abstract

The invention discloses an electronic mediator type enzymatic electrochemical immunodetection method of p53 protein. The electronic mediator type enzymatic electrochemical immunodetection method specifically comprises the following steps: preparing an immune molecule immobilized active interface; constructing and optimizing an electronic mediator type enzymatic electrochemical immunoassay system; and performing quantitative detection on p53 protein. The method specifically comprises the following steps: performing electrostatic spinning and performing electropolymerization on thionine to obtain a PA6-MWCNTs-PTH immune molecule immobilized active interface, constructing the electronic mediator type enzymatic electrochemical immunoassay system on the PA6-MWCNTs-PTH interface by virtue of sandwich immunoreaction, performing relevant parameter optimization, and performing quantitative detection on p53 protein by using the constructed immunoassay system. The detection method disclosed by the invention has the characteristics of high sensitivity, high selectivity, high stability, regeneration performance, wide linear range, low detection limit and the like.

Description

technical field [0001] The invention relates to a method for detecting p53 protein. Background technique [0002] p53 protein is the expression product of p53, the highest human tumor-related gene, and is the most representative and important tumor-related protein. The amount of p53 protein in normal human cells is very low. These low-level p53 proteins are easily hydrolyzed and have a short half-life of only 10-20 minutes, which is generally difficult to detect. The traditional detection methods of p53 protein are immunoassays, such as radioimmunoassay (RIA), immunohistochemistry (IHC) and enzyme-linked immunoassay (ELISA). The RIA method has radioactive pollution and is not easy to be promoted in clinic. The IHC method is only applied to the study of p53 protein expression in tissue samples and cell samples, and its operation procedure is complicated and takes a long time. ELISA is currently the most commonly used method. It can be used for both qualitative and quantita...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/68G01N27/48G01N33/531G01N2333/4748
Inventor 王晓英
Owner SOUTHEAST UNIV
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