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Kit for detecting carcino-embryonic antigen, detection method and application thereof

A carcinoembryonic antigen and detection method technology, applied in the field of electrochemical detection, can solve the problems of easy poisoning of the working electrode, application limitations of electrochemical immunosensors, difficulty in meeting the high sensitivity and precision of CEA, and shorten the immune reaction time, High sensitivity, reduce the effect of poisoning

Inactive Publication Date: 2019-08-16
HUBEI UNIV FOR NATITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the electrochemical signal is easily affected by the electrode activity, and the working electrode is easily poisoned in complex samples, the application of electrochemical immunosensors in actual complex samples is often limited.
In addition, current immunoassay methods are difficult to meet the high sensitivity and precision required for CEA detection

Method used

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  • Kit for detecting carcino-embryonic antigen, detection method and application thereof
  • Kit for detecting carcino-embryonic antigen, detection method and application thereof
  • Kit for detecting carcino-embryonic antigen, detection method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0068] Preparation example 1 Preparation of nano-gold bifunctional probe

[0069] Such as figure 1 As shown, the preparation process of nano-gold bifunctional probe is as follows:

[0070] (1) Preparation of gold nanoparticles (GSH-AuNPs) with glutathione as a ligand

[0071] The gold nanoparticles used in the experiment were prepared by a quasi-biosynthetic method. The gold nanoparticles use glutathione as a ligand, which has very good colloidal stability, and its surface is rich in carboxyl and amino groups, which can be directly used for modification and coupling. The specific operation steps are as follows:

[0072] Add 0.10 mL of glutathione (GSH, 0.24 mol / L) to 1 mL of 1% chloroauric acid, stir at room temperature for 2 minutes, then slowly add 1 mol / L NaOH dropwise to the solution until the pH of the solution is between 2.5 -3.0, a large number of light yellow precipitates appeared in the solution. Then use a centrifuge to centrifuge at a speed of 8000 rpm for 1 mi...

preparation example 2

[0080] Preparation Example 2 Preparation of Immunomagnetic Beads

[0081] The CEA capture antibody (i.e. primary antibody, also referred to as Ab for short) was used for EDC / NHS activation 1 ) is covalently coupled to the surface of carboxyl-functionalized magnetic beads to obtain immunomagnetic beads. The specific operation process is as follows:

[0082] Take 10 μL carboxyl-functionalized superparamagnetic microspheres (30 mg / mL, 300 nm) and wash twice with 0.1 mol / L PBS at pH 6.0. Then it was dispersed into 1 mL of PBS (0.1 mol / L pH 6.0) solution containing 0.1 mol / L LEDC and 0.1 mol / L NHS, and activated on a constant temperature shaker at 37°C with a rotation speed of 160 rpm for 60 min. After washing four times with 0.1mol / L PBS with a pH of 7.2, resuspend in 400 μL of PBS buffer (0.1mol / L, pH 7.2), and add 10 μL of 0.55 mg / mL CEA capture antibody (Ab 1 ), and the reaction was incubated on a shaker for 10 h. Then wash four times with 0.1mol / L PBS with pH 7.2 to remove...

preparation example 3

[0083] Processing and Modification of Preparation Example 3 Magnetic Glassy Carbon Electrode

[0084] A magnetic glassy carbon electrode (M-GCE) with a diameter of 3 mm was polished to a mirror surface on the suede with 0.3 μm and 0.05 μm Al2O3 powder, and then ultrasonically cleaned with absolute ethanol and ultrapure water for half a minute, and a large amount of Rinse with ultrapure water and place in a phosphate buffer solution (0.1M, pH7.0) containing 1mM potassium ferricyanide and 1mM potassium ferrocyanide, and perform cyclic voltammetry scanning in the potential range of -0.2-0.6V , when the potential difference of the oxidation-reduction peak of potassium ferricyanide is below 80mV, it indicates that the electrode surface is in good condition. Then immerse the cleaned magnetic glassy carbon electrode in an acetonitrile solution containing 0.1mol / L ethylenediamine and 0.1mol / L tetrabutylammonium perchlorate (TBAP), and use platinum wire as the counter electrode and ref...

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Abstract

The invention relates to the field of electrochemical detection and discloses a kit for detecting a carcino-embryonic antigen, a detection method and application thereof. The kit disclosed by the invention comprises a nanogold dual-function probe and immunomagnetic beads, wherein the nanogold dual-function probe comprises nanogold particles, a second antibody and a detection marker, wherein the second antibody and the detection marker are connected to the nanogold particles; the immunomagnetic beads comprise magnetic beads and a first antibody connected to the magnetic beads; and the first antibody and the second antibody are respectively and independently an antibody of an anti-cancer embryo antigen. The kit and the detection method in the invention have the advantages of being high in sensitivity, high in detection speed and good in repeatability.

Description

technical field [0001] The invention relates to the field of electrochemical detection, in particular to a kit for detecting carcinoembryonic antigen, a detection method and an application thereof. Background technique [0002] Cancer is one of the main causes of human death, and early clinical diagnosis can effectively reduce the mortality of patients. Tumor markers refer to specific substances present in tumor cells themselves or secreted by tumor cells, which can reflect the existence and growth of tumors. Carcinoembryonic antigen (CEA) is one of the most widely used tumor markers. According to the content of CEA in serum, it can be used for clinical research and early diagnosis of cancer. Therefore, the sensitive detection of CEA has attracted widespread attention of scientists. So far, researchers have developed a series of analytical techniques for highly sensitive detection of CEA, such as fluorescence analysis (FIA), enzyme-linked immunosorbent assay (ELISA), electr...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/574
CPCG01N33/54326G01N33/54346G01N33/57473
Inventor 齐宝平尚冰冰
Owner HUBEI UNIV FOR NATITIES
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