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Thrombin aptamer sensor based on Ti3C2 and preparation method of thrombin aptamer sensor

An aptamer sensor and thrombin technology, 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., and achieve the effect of fast reaction, easy storage, and simple synthesis

Active Publication Date: 2020-03-24
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Thrombin aptamer sensor based on Ti3C2 and preparation method of thrombin aptamer sensor
  • Thrombin aptamer sensor based on Ti3C2 and preparation method of thrombin aptamer sensor
  • Thrombin aptamer sensor based on Ti3C2 and preparation method of thrombin aptamer sensor

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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. Ti 3 C2 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 thrombin aptamer sensor based on Ti3C2 and a preparation method of the thrombin aptamer sensor which belong to the technical field of biological analysis and detection, andsolve the technical problems that an antibody used in a traditional thrombin detection method is difficult to prepare and store, and the related reactions are complex and time-consuming. The thrombinaptamer sensor based on Ti3C2 is composed of a single layer of Ti3C2 and a fluorescein-labeled thrombin aptamer combined to the surface of the Ti3C2, and the sequence of the fluorescein labeled thrombin aptamer is 5 '-FAM-GGT TGG TGT TGT TGG-3'. According to the invention, the thrombin sensor is constructed by adopting a Ti3C2 material, so that the application of Ti3C2 in the biological field is broadened, and an excellent nano detection platform is constructed; the thrombin aptamer is used as a thrombin recognition agent, and compared with a traditional antibody, the thrombin aptamer is low in price, is easy and convenient to synthesize, is easy to store and is rapid in reaction. The Ti3C2-based thrombin aptamer sensor prepared by the preparation method provided by the invention 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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IPC IPC(8): G01N21/64
CPCG01N21/6428G01N21/6486G01N2021/6417G01N2021/6432
Inventor 王晓峰崔洪源
Owner JILIN UNIV
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