Based on ti 3 c 2 Thrombin aptamer sensor and preparation method thereof

An aptamer sensor and thrombin technology, which is applied in instruments, scientific instruments, fluorescence/phosphorescence, etc., can solve the problems of complex and time-consuming reactions, difficult preparation and storage of antibodies, etc.

Active Publication Date: 2021-07-06
JILIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the disadvantages of these methods are that the antibodies used are difficult to prepare and store, and the related reactions are complicated and time-consuming

Method used

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  • Based on ti <sub>3</sub> c <sub>2</sub> Thrombin aptamer sensor and preparation method thereof
  • Based on ti <sub>3</sub> c <sub>2</sub> Thrombin aptamer sensor and preparation method thereof
  • Based on ti <sub>3</sub> c <sub>2</sub> Thrombin aptamer sensor and preparation method thereof

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preparation example Construction

[0039] The present invention also provides a Ti-based 3 C 2 The preparation method of the thrombin aptamer sensor comprises the following steps:

[0040] (1) Monolayer Ti 3 C 2Preparation of:

[0041] Ti is prepared by sintering a mixture of titanium powder, aluminum powder, and graphite under argon 3 AlC 2 , will Ti 3 AlC 2 The powder was added to the LiF / HCl solution for etching under continuous stirring. After the reaction was complete, the acidic mixture was washed with ultrapure water by repeated centrifugation until the pH of the mixture exceeded 5. Finally, the slurry was sonicated and centrifuged to obtain a monolayer Ti 3 C 2 solution;

[0042] (2) Bind fluorescein-labeled thrombin aptamer to Ti 3 C 2 surface:

[0043] Add the fluorescein-labeled thrombin aptamer to the Ti prepared in step (1) at room temperature 3 C 2 In solution, after the reaction is complete, the composition is based on Ti 3 C 2 Thrombin aptamer sensor.

[0044] Preferably, the m...

Embodiment 1

[0062] 1) Monolayer Ti 3 C 2 Synthesis: Ti was prepared by sintering a mixture of 7.368g titanium powder, 1.523g aluminum powder and 1.109g graphite under argon at 1650°C for 2 hours 3 AlC 2 . Fired Ti 3 AlC 2 The powder was ground in a mortar and passed through a 400-mesh sieve. 1gTi 3 AlC 2 It was added into a 12M LiF / 9M HCl solution for etching under continuous stirring, and reacted at room temperature for 24 hours. After the etching was completed, the centrifugation was repeated, and the mixture was washed with ultrapure water at 8000 rpm for 5 minutes per cycle until the pH of the mixture exceeded 5. Finally, the slurry was sonicated for more than 10 min and centrifuged at 3500 rpm for 1 h to obtain monolayer Ti 3 C 2 solution. To confirm Ti 3 C 2 concentration, filter 10mL Ti with a cellulose membrane with a pore size of 0.22 μm 3 C 2 Dispersions. Ti 3 C 2 After drying, the Ti 3 C 2 The film was peeled off and weighed, and Ti was calculated as 3 C 2...

Embodiment 2

[0067] 1) Monolayer Ti 3 C 2 Synthesis of Ti: Ti was prepared by sintering a mixture of 3.684 g titanium powder, 0.762 g aluminum powder and 0.555 g graphite at 1650 °C under argon for 2 hours 3 AlC 2 . Fired Ti 3 AlC 2 The powder was ground in a mortar and passed through a 400-mesh sieve. 1gTi 3 AlC 2 Add it into 8M LiF / 6M HCl solution under continuous stirring for etching, and react at room temperature for 24 hours. After the etching was completed, the centrifugation was repeated, and the mixture was washed with ultrapure water at 8000 rpm for 5 minutes per cycle until the pH of the mixture exceeded 5. Finally, the slurry was sonicated for more than 10 min and centrifuged at 3500 rpm for 1 h to obtain monolayer Ti 3 C 2 solution. To confirm Ti 3 C 2 concentration, filter 10mL Ti with a cellulose membrane with a pore size of 0.22 μm 3 C 2 Dispersions. Ti3 C 2 After drying, the Ti 3 C 2 The film was peeled off and weighed, and Ti was calculated as 3 C 2 co...

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Abstract

The invention relates to a Ti-based 3 C 2 A thrombin aptamer sensor and a preparation method thereof belong to the technical field of biological analysis and detection. It solves the technical problems that the antibodies used in traditional thrombin detection methods are difficult to prepare and store, and the related reactions are complicated and time-consuming. The Ti-based 3 C 2 The thrombin aptamer sensor is composed of monolayer Ti 3 C 2 and its binding to Ti 3 C 2 The composition of the fluorescein-labeled thrombin aptamer on the surface; the sequence of the fluorescein-labeled thrombin aptamer is: 5′-FAM-GGT TGG TGT GGT TGG-3′. The present invention uses Ti 3 C 2 Material builds thrombin sensor, broadens Ti 3 C 2 In the application in the biological field, an excellent nano-detection platform has been constructed; the thrombin aptamer is used as the thrombin recognition agent, which is cheap, easy to synthesize, easy to store, and fast in response compared with traditional antibodies. The invention provides the Ti-based 3 C 2 The thrombin aptamer sensor has the characteristics of high specificity and high sensitivity, and the detection limit of thrombin is 2.5×10 ‑11 mol / L.

Description

technical field [0001] The invention belongs to the technical field of biological analysis and detection, and in particular relates to a Ti-based 3 C 2 Thrombin aptamer sensor and preparation method thereof. Background technique [0002] Thrombin is a Na + Activated allosteric serine protease that functions as a central protease in the coagulation cascade. Thrombin and the inactive prothrombin play crucial roles in physiology and pathology. Thrombin has been linked to many diseases such as blood clots, Alzheimer's disease, central nervous system damage and cancer. Therefore, highly sensitive and specific thrombin detection methods are essential for the research and clinical diagnosis of these diseases. [0003] Currently, a variety of methods have been developed for the detection of thrombin, including coagulation-based assays, enzyme activity-based assays, and immunoassays. However, the disadvantages of these methods are that the antibodies used are difficult to prepa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6486G01N2021/6417G01N2021/6432
Inventor 王晓峰崔洪源
Owner JILIN UNIV
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