Three-dimensional graphene based proportional signal amplification aptasensor, preparation method and application thereof

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

Active Publication Date: 2018-09-04
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the existing detection methods still have some shortcomings,

Method used

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  • Three-dimensional graphene based proportional signal amplification aptasensor, preparation method and application thereof
  • Three-dimensional graphene based proportional signal amplification aptasensor, preparation method and application thereof
  • Three-dimensional graphene based proportional signal amplification aptasensor, preparation method and application thereof

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 three-dimensional graphene based proportional signal amplification aptasensor. The method adopts a gold nano-three-dimensional graphene compound as the modified electrode substrate material, then grafts ferrocene-labeled mucin aptamer to gold nanoparticles on a three-dimensional graphene surface, and employs bovine serum albumin to close non-specific sites on a sensing interface, and when the ferrocene-labeled aptamer captures mucin to an electrode surface, a methyl blue labeled mucin aptamer-gold nano compound is employed for co-incubation, thus obtaining the aptasensor for electrochemical detection. According to the invention, the aptamer is employed to construct a sensing interface, and is conducive to enhancing the stability and selectivity of the sensor; at the same time, the two electroactive substances ferrocene and methyl blue are introduced into the sensing system to play an internal reference role and a signal amplification role respectively. The construction method of the proportional signal amplification aptasensor has the advantages of high sensitivity, good stability, simple method, high cost-effectiveness, etc.

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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IPC IPC(8): G01N27/327G01N27/48
CPCG01N27/3278G01N27/48G01N27/3275
Inventor 梁琼麟杨少红
Owner TSINGHUA UNIV
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