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Method for detection of antigen content in samples based on immunohistochemical space electrochemistry

A technology for antigen content and sample detection, applied in the biological field, can solve problems such as high technical requirements, low sensitivity, and complicated operation, and achieve the effect of low technical requirements, improved sensitivity, and simple operation

Active Publication Date: 2021-11-09
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to its complicated operation: high technical requirements for pathological technicians; relatively low sensitivity: due to optical limitations, sometimes trace antigens cannot be accurately observed; cannot be quantified: immunohistochemical films can only be relatively quantitative, and several people are required Experts give a unified conclusion, there are many human factors; time-consuming: due to the many procedures of immunohistochemistry, it takes 2 to 3 days to give clinical patients a prepared pathological report, which is likely to affect the progress of subsequent treatment
[0005] Therefore, immunohistochemical detection in the prior art still has shortcomings such as incomplete quantification and complicated operation.

Method used

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  • Method for detection of antigen content in samples based on immunohistochemical space electrochemistry
  • Method for detection of antigen content in samples based on immunohistochemical space electrochemistry
  • Method for detection of antigen content in samples based on immunohistochemical space electrochemistry

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Experimental program
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Effect test

Embodiment 1

[0078] Electrode modification flow chart as figure 1 As indicated, take 30mM HAuCl 4 , 0.1M Na 2 SO 4 , in chloroauric acid and sodium sulfate solution, use chronoamperometry to electroplate the surface of the electrode to obtain dendritic gold, and the morphology of the obtained dendritic gold is as follows figure 2 shown. Using cyclic voltammetry and electrochemical impedance method to compare the current resistance value of the electrode before and after dendritic gold modification, such as image 3 shown.

[0079] Dissolve the conductive polymer monomer (ethylenedioxythiophene in this example) and tetrabutylammonium perchlorate in anhydrous acetonitrile, pass through nitrogen, remove the air in the solution, and use cyclic voltammetry to dissolve the conductive polymer electroplating on the surface of the dendritic gold to obtain an electrode modified with the dendritic gold and the conductive polymer.

[0080] Among them, the concentrations of conductive polymer mo...

Embodiment 2

[0089] Take 30mM HAuCl 4 , 0.1M Na 2 SO 4 , Dendritic gold was obtained after electroplating by chronoamperometry, and conductive polymer monomers such as ethylenedioxythiophene, thiopheneacetic acid, and pyrrole were dissolved in anhydrous acetonitrile, and nitrogen gas was introduced to remove the solution In neutral air, polymers were electroplated on dendritic gold surfaces using cyclic voltammetry. In a solution containing 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, ferrocene was added to co-culture with specific antibodies, and then the to-be-detected The slides of the frozen section tissue were immersed in the solution, co-incubated, the slides were removed, and the electrochemical signals of unbound antibodies in the solution were detected. The process is as follows: Figure 7 shown.

Embodiment 3

[0091] Take 30mM HAuCl 4 , 0.1M Na 2 SO 4, dendritic gold was obtained after electroplating the electrode surface using chronoamperometry. Dissolve conductive polymer monomers such as ethylenedioxythiophene, thiopheneacetic acid and pyrrole with tetrabutylammonium perchlorate in acetonitrile, pass through nitrogen, remove the air in the solution, and use cyclic voltammetry to electroplate the conductive polymer on the A dendritic gold surface, and an electrode modified with a dendritic gold and a conductive polymer is obtained. The electrochemical signal (ferrocene), in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide in acetonitrile and Specific antibodies were co-incubated to obtain antibodies modified by electrochemical signals. Add the antibody modified by the electrochemical signal to the cells to be detected, and incubate together to carry out the binding of the antigen and antibody, centrifuge to remove the unbound...

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Abstract

The invention relates to a method for detecting antigen content in a sample based on immunohistochemical spatial electrochemical detection, comprising the following steps: modifying electrochemical signals (ferrocene / quantum dots, etc.) For the bound antibody, an electrochemical instrument is used to detect the electrochemical signal in the solution, and the strength of the signal reflects the content of the antigen in the cancer tissue / cell. Compared with the prior art, the present invention can realize the simultaneous detection of different antigens by using different electrochemical signals to modify antibodies; through the determination of the standard curve, the electrochemical signal value directly corresponds to the number of antigens, realizing full quantitative detection; Compared with immunohistochemistry, the detection steps are simple to operate and require less technical skills for the operator; the modified electrode can greatly improve the sensitivity and realize the detection of micro-antigens.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for detecting antigen content in a sample based on immunohistochemical space electrochemistry. Background technique [0002] Immunohistochemical detection, referred to as immunohistochemical detection, is the application of the basic principle of immunology - antigen-antibody reaction, that is, the principle of specific combination of antigen and antibody, through chemical reaction to make the chromogenic agent (fluorescein, enzyme, metal, etc.) of the labeled antibody ions, isotopes, etc.) to determine tissue cell antigens (polypeptides, proteins, pathogens, etc.) . According to the types of markers, immunohistochemistry can be roughly divided into immunofluorescence cytochemistry, immunoenzyme cytochemistry, and immunocolloidal gold techniques. [0003] In conventional pathological diagnosis of tumors, in some cases it is difficult to make an accurate diagnos...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N27/48
CPCG01N27/48G01N33/54346
Inventor 王平秦洁玲王佳
Owner TONGJI UNIV
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