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Preparation method and application of electrochemiluminescence aptamer sensor for detecting tumor marker MUC1

An aptamer sensor and tumor marker technology, which is applied in the direction of chemiluminescence/bioluminescence, and analysis by making materials undergo chemical reactions, can solve the problems of undisclosed electrochemiluminescence aptamer sensors, etc., and achieve high sensitivity, The effect of high sensitivity and structural design improvement

Inactive Publication Date: 2015-05-27
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no disclosure of any electrochemiluminescent aptamers for detecting tumor marker MUC1 based on magnetic graphene oxide, electrochemiluminescent bodies and nucleic acid aptamers with specific recognition ability for tumor marker MUC1 at home and abroad. Related research reports on the preparation method and application of the sensor

Method used

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  • Preparation method and application of electrochemiluminescence aptamer sensor for detecting tumor marker MUC1
  • Preparation method and application of electrochemiluminescence aptamer sensor for detecting tumor marker MUC1
  • Preparation method and application of electrochemiluminescence aptamer sensor for detecting tumor marker MUC1

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specific Embodiment 1

[0034] A method for preparing an electrochemiluminescence aptasensor for detecting tumor marker MUC1, comprising the following steps:

[0035] (1) Synthesis of multifunctional graphene oxide materials, the preparation process is as follows figure 1 shown

[0036] a. Synthesis of magnetic graphene oxide: Add 40 mL of 1 mg / mL graphene oxide (GO) into a clean three-necked flask, sonicate for 1 h (1 h for sonication, the power generally used is about 200W); Add 25 mL of 0.06 mol / L ferric chloride solution and 25 mL of 0.12 mol / L ferric chloride solution under nitrogen atmosphere, stir and reflux at 80°C, and slowly add 12 mL of 25% ammonia water dropwise to the pH of the solution = 10.5, reflux treatment for 4 h; after cooling, wash with secondary water until the pH of the solution = 7, and set the volume to 50 mL to obtain a magnetic graphene oxide material solution, which is stored at 4 °C for use;

[0037] b. Take 200 μL of the magnetic graphene oxide material solution obtain...

specific Embodiment 2

[0040] With above-mentioned embodiment 1, its main difference is:

[0041] In step a of step (1): add 30 mL of 1 mg / mL graphene oxide into a clean three-necked flask, sonicate for 1 h; then add 20 mL of 0.07 mol / L ferrous chloride under nitrogen atmosphere solution and 20 mL of 0.15 mol / L ferric chloride solution, stirred and refluxed at 70°C, while slowly adding 10 mL of 30% ammonia water to the pH of the solution = 10, and refluxed for 5 h; after cooling, washed with secondary water To the pH of the solution = 7, constant volume to 50 mL, to obtain a magnetic graphene oxide material solution;

[0042] In step b of step (1), take the magnetic graphene oxide material solution and sonicate it in a glass bottle, then add 200 μL of coupling reagent, and at the same time add dilute hydrochloric acid solution to pH = 4, incubate with shaking for 1 h, and wash with water twice 3 times; then add 30 μL 0.1 mol / L electrochemiluminescent solution (the solute is N-(4-aminobutyl)-N-ethyl...

specific Embodiment 3

[0044] With above-mentioned embodiment 1, its main difference is:

[0045] In step a of step (1): add 50 mL of 1 mg / mL graphene oxide into a clean three-necked flask, sonicate for 1 h; then add 30 mL of 0.05 mol / L ferrous chloride under nitrogen atmosphere solution and 30 mL of 0.10 mol / L ferric chloride solution, stirred and refluxed at 90°C, while slowly adding 15 mL of 20% ammonia water to the pH of the solution = 11, and refluxed for 3 h; after cooling, washed with secondary water To the pH of the solution = 7, constant volume to 50 mL, to obtain a magnetic graphene oxide material solution;

[0046] In step b of step (1), take the magnetic graphene oxide material solution and sonicate it in a glass bottle, then add 400 μL of coupling reagent, mix well, and at the same time add dilute hydrochloric acid solution to pH = 6, and incubate with shaking for 1 h. Wash 3 times with secondary water; then add 50 μL 0.001 mol / L luminol electrochemiluminescent solution and 50 μL 10 μm...

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Abstract

The invention discloses a preparation method and an application of an electrochemiluminescence aptamer sensor for detecting a tumor marker MUC1. The preparation method is characterized by comprising a step of synthesizing magnetic graphene oxide; simultaneously marking an electrochemiluminescence body and an aptamer on the surface of the magnetic graphene oxide material so as to obtain a multifunctional graphene oxide material; and a step of finally dropping and applying 5-10 mu L of multifunctional graphene oxide material to the surface of a magnetic glassy carbon electrode so as to obtain an electrochemiluminescence aptamer sensor for detecting the tumor marker MUC1. The electrochemiluminescence aptamer sensor can be used for determining the concentration of the tumor marker MUC1 in a to-be-detected sample on the basis of a quantitative relation between electrochemiluminescence intensity and the concentration of a tumor marker MUC1 solution, and the sensor has the advantages of being simple and rapid in detection method, sensitive and accurate in detection result, and high in specificity.

Description

technical field [0001] The present invention relates to an electrochemiluminescent aptamer sensor, in particular to a preparation method and application of an electrochemiluminescent aptamer sensor for detecting tumor marker MUC1. Background technique [0002] In recent years, the incidence of cancer in the world has become increasingly serious, and the number of patients dying of malignant tumors has increased year by year. Early diagnosis and treatment of tumors is the key to reducing mortality. At present, the clinical diagnosis of cancer is mainly realized by monitoring the content of tumor markers in the human body. . Tumor markers refer to a class of biochemical substances produced by tumor cells themselves or by the host’s response to tumors in the body during tumorigenesis and proliferation, including tumor-associated protein markers, hormone markers, and enzyme markers. , gene markers and embryonic antigens can be detected in tumor tissue, blood, body fluids or exc...

Claims

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Application Information

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IPC IPC(8): G01N21/76
Inventor 李温柔郭智勇张宁陈贝贝沙玉红武琳
Owner NINGBO UNIV
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