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A three-dimensional graphene-based proportional signal amplification aptamer sensor and its preparation method and application

A signal amplification and graphene-based technology, applied in instruments, scientific instruments, measuring devices, etc., can solve the problems of high detection cost, poor selectivity, and low sensitivity, and achieve good biocompatibility, improved accuracy, and improved sensitivity Effect

Active Publication Date: 2019-11-22
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing detection methods still have some shortcomings, such as low sensitivity, poor selectivity, high detection cost, etc.

Method used

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  • A three-dimensional graphene-based proportional signal amplification aptamer sensor and its preparation method and application
  • A three-dimensional graphene-based proportional signal amplification aptamer sensor and its preparation method and application
  • A three-dimensional graphene-based proportional signal amplification aptamer sensor and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Embodiment 1, the preparation of gold nano-three-dimensional graphene composite material (Au-RGO)

[0076] (1) ultrasonically disperse 5 mg of graphene oxide in 1 ml of ultrapure water for 2 hours to obtain a uniformly dispersed graphene oxide suspoemulsion;

[0077] (2) Dilute the graphene oxide suspoemulsion in (1) to 6 mL, then add 200 μL tetrachloroauric acid solution (1%, w / w) and mix well; then add 25 mg polyethylene glycol to 4 mL ultrapure water, After it is fully dissolved, it is added into the mixed solution of graphene oxide and tetrachloroauric acid, and ultrasonically treated for 1 hour to make it fully mixed.

[0078] (3) The mixed solution in (2) was transferred to a 25mL hydrothermal reaction device, reacted in an oven at 160°C for 12h, and then cooled to room temperature. After the reaction product was taken out, it was centrifuged and washed three times with ultrapure water, and the obtained black precipitate was retained.

[0079] (4) Add 8 mL of ul...

Embodiment 2

[0081] Embodiment 2, the preparation of gold nano-three-dimensional graphene composite material (Au-RGO)

[0082] (1) Ultrasonic dispersion of 4 mg graphene oxide in 1 ml ultrapure water for 1 h to obtain a uniformly dispersed graphene oxide suspoemulsion;

[0083] (2) Dilute the graphene oxide suspoemulsion in (1) to 8 mL, then add 150 μL tetrachloroauric acid solution (1%, w / w) and mix well; then add 20 mg polyethylene glycol to 2 mL ultrapure water, After it is fully dissolved, it is added to the mixed solution of graphene oxide and tetrachloroauric acid, and ultrasonically treated for 0.5 h to make it fully mixed.

[0084] (3) Transfer the mixed solution in (2) into a 25mL hydrothermal reaction device, react in an oven at 170°C for 15h, and then cool to room temperature. After the reaction product was taken out, it was centrifuged and washed three times with ultrapure water, and the obtained black precipitate was retained.

[0085] (4) Add 5 mL of ultrapure water to the pr...

Embodiment 3

[0086] Example 3, preparation of gold nanoparticles and methylene blue-labeled MUC1 aptamer-gold nanocomposites:

[0087] (1) get 80mL concentration and be that the tetrachloroauric acid solution of 1mM is in the heating reflux device, be heated to vigorous boiling under the magnetic sub-stirring condition;

[0088] (2) Rapidly add 10 mL of trisodium citrate solution with a concentration of 38.8 mM to (1), and then heat to reflux for 10 min;

[0089] (3) After stopping the heating, cool to room temperature with stirring, and transfer the obtained wine-red gold nanometer into a brown reagent bottle and place it in a 4°C refrigerator for later use.

[0090] Such as image 3 As shown, the diameter of the synthesized uniform gold nanoparticles is about 15nm.

[0091] (4) Take 1 mL of the gold nanoparticles prepared in (3), and add 150 μL of 50 μM methylene blue-labeled MUC1 aptamer solution to it, and incubate at 25° C. for 16 h with slight shaking on a shaker;

[0092] (5) Tra...

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Abstract

The invention discloses a preparation method of a proportional signal amplification aptamer sensor based on three-dimensional graphene. In this method, the gold nano-3D graphene composite is used as the modified electrode substrate material, and then ferrocene-labeled mucin aptamers are grafted onto the gold nanoparticles on the surface of the 3D graphene, and bovine serum albumin is used to control the non-specific properties of the sensing interface. The heterosexual sites were blocked. After the ferrocene-labeled aptamer captures mucin on the electrode surface, methylene blue-labeled mucin aptamer-gold nanocomposite is incubated with it and then used for electrochemical detection. The present invention adopts the aptamer to construct the sensing interface, which is beneficial to enhance the stability and selectivity of the sensor; at the same time, two electroactive substances, ferrocene and methyl blue, are introduced into the sensing system, which can respectively play the role of internal reference and Signal amplification. The construction method of the proportional signal amplification aptamer sensor has the advantages of high sensitivity, good stability, simple method, high cost-effectiveness and the like.

Description

technical field [0001] The invention relates to a three-dimensional graphene-based proportional signal amplification aptamer sensor and a preparation method and application thereof, and belongs to the technical field of novel functionalized nanomaterials and electrochemical biosensing. Background technique [0002] Tumor is a major disease that poses a serious threat to human life and health. The detection of tumor markers plays a key role in the timely detection and early treatment of tumors. Mucin 1 (Mucin1, MUC1) is a high molecular weight (>200KD) glycoprotein, because of its abnormal expression in tumor tissues, it has been used in the early diagnosis and biological treatment of tumors. [0003] MUC1 is abnormally expressed on the surface of most malignant tumor epithelial cells, such as breast cancer, lung cancer, pancreatic cancer, ovarian cancer, prostate cancer, bladder cancer, colorectal cancer and so on. In normal human epithelial cells, the expression of MUC...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3278G01N27/48G01N27/3275
Inventor 梁琼麟杨少红
Owner TSINGHUA UNIV
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