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A sensor based on molybdenum disulfide and its preparation method and application

A molybdenum disulfide and sensor technology, applied in the field of biomedicine, can solve the problems of preventing detection sensitivity and low cost, and achieve the effects of easy acquisition, low cost, and avoiding false signals

Active Publication Date: 2018-02-27
镇江永晨科技有限公司
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0003] MoS 2 The preparation is simple and the cost is relatively low, so it has been widely used in biological detection, and MoS 2 When interacting with DNA, there is an important feature that single-stranded DNA can be adsorbed to MoS through the π-π effect. 2 On the surface, aptamer is a single-stranded DNA that can specifically bind to thrombin. When thrombin specifically binds to aptamer, aptamer will detach from MoS 2 On the surface, based on this feature, some researchers have specifically detected thrombin through fluorescently labeled aptamers (Xiang, X., Shi, J.B., Huang, F.H., Zheng, M.M., Deng, Q.C.Xu, J.Q., MoS 2 nanosheet-based fluorescent biosensor for protein detection via terminal protection of small-molecule-linked DNA and exonuclease III-aided DNA recycling amplification 2015. Biosensors and Bioelectronics 74, 227–232.), but this method has a disadvantage: aptamer and the detected thrombin can Non-specific adsorption to MoS 2 surface, thereby preventing an increase in detection sensitivity

Method used

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  • A sensor based on molybdenum disulfide and its preparation method and application
  • A sensor based on molybdenum disulfide and its preparation method and application
  • A sensor based on molybdenum disulfide and its preparation method and application

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Experimental program
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Embodiment 1

[0030] (1) Preparation of MoS 2 : Prepared according to the method of Eda (Eda, et.al.Photoluminescence fromchemically exfoliated MoS 2 .Nano Letters, 2011, 11, 5111-5116.):

[0031] 3g natural MoS 2 The crystals were immersed in argon-filled 3mL 1.6M n-butyllithium flask for 2 days for lithium intercalation; then, filtered and rinsed with n-hexane to produce Li x MoS 2 Remove impurities. Li x MoS 2 Sonicated in ultrapure water for 1 h, the obtained suspension was centrifuged, rinsed with water, and then filtered with a mixed cellulose ester membrane to obtain a precipitate denatured in an argon-protected glove box at 300 ° C for 1 hour to obtain MoS 2 Soluble in water, ultrasonic 2h standby.

[0032] (2) Preparation of gold nanoparticles: Gold nanoparticles were prepared with reference to the method of Frens (Frens G, Controlled Nucleation for the Regulation of the Particle Size in Monodisperse Gold Suspensions,. Nature PhySci, 1973, 241, 20-22), and the obtained AuNP av...

Embodiment 2

[0041] Steps (1)-(4) are the same as in Example 1.

[0042] (5) Fluorescence quenching: adding nucleotide aptamers to MoS 2- In the aqueous solution of AuNP, Tween80 was added to obtain a mixed solution. After standing at room temperature for 30 minutes, the mixed solution was heated in a water bath at 70°C for 2.5 hours, centrifuged at 10000r for 15 minutes, and then the precipitate was dispersed in 0.1M PBS. That is, the preparation of MoS based on the covalent binding of nucleotide aptamers 2- AuNP sensor for detecting thrombin; MoS 2- AuNPs are capable of quenching the fluorescence of nucleotide aptamer-end-labeled FAMs.

[0043] Among them, MoS 2- The concentration of nucleotide-containing aptamers in the aqueous solution of AuNP was 10 nM, and the MoS contained 2- The concentration of AuNP is 60 μg / mL;

[0044] The MoS added to the nucleotide aptamer 2- The aqueous solution of AuNP is 1 mL, and the volume of Tween80 added is 10 μL; the mass percentage of Tween80 is...

Embodiment 3

[0048] Steps (1)-(4) are the same as in Example 1.

[0049] (5) Fluorescence quenching: adding nucleotide aptamers to MoS 2- In the aqueous solution of AuNP, Tween80 was added to obtain a mixed solution. After standing at room temperature for 30 minutes, the mixed solution was heated in a water bath at 70°C for 2.5 hours, centrifuged at 10000r for 15 minutes, and then the precipitate was dispersed in 0.1M PBS. , which is prepared as a covalently bound MoS based on nucleotide aptamers 2- AuNP sensor for detecting thrombin; MoS 2- AuNPs are capable of quenching the fluorescence of nucleotide aptamer-end-labeled FAMs.

[0050] Among them, MoS 2- The concentration of nucleotide-containing aptamers in the aqueous solution of AuNP was 10 nM, and the MoS contained 2- The concentration of AuNP is 60 μg / mL;

[0051] The MoS added to the nucleotide aptamer 2- The AuNP solution is 1 mL, and the volume of Tween80 added is 10 μL; the mass percentage of Tween80 is 10%;

[0052] The f...

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Abstract

The invention relates to a molybdenum disulfide based sensor as well as a preparation method and an application thereof and belongs to a protein detection method in the biomedical field. According to the technical scheme, the method mainly comprises steps as follows: firstly, an MoS2-AuNP compound is prepared; a nucleotide aptamer of thrombin is synthesized; the synthesized nucleotide aptamer is added to an aqueous solution of MoS2-AuNP, Tween 80 is added, a mixed solution is obtained, reacts for 30 min at the room temperature, then is heated for 2.5 h in a water bath with the temperature of 70 DEG C and is centrifuged at the speed of 10,000 r for 15 min, and then, precipitates are dispersed in a 0.1 M PBS (phosphate buffer solution); and the sensor is prepared. The characteristic that MoS2-AuNP can quench fluorescence is sufficiently utilized, an FAM (carboxyfluorescein)-marked nucleotide aptamer sequence is combined with thrombin specificity, trace detection of thrombin can be realized in a high-sensitivity, rapid and low-cost manner through quenching and recovery of FAM fluorescence, and the actual LOD (limit of detection) of the MoS2-AuNP-nucleotide aptamer sensor to thrombin is increased to 0.4 pM due to addition of Tween 80.

Description

technical field [0001] The present invention relates to a sensor based on molybdenum disulfide and its preparation method and application, in particular to a thiol-based nucleotide aptamer (aptamer) covalently bound to molybdenum disulfide-gold nanocomposites (MoS 2- AuNP) sensor and application thereof belong to the protein detection method in the field of biomedicine. Background technique [0002] Human thrombin is the main effector protease of the coagulation chain reaction. In addition to its ability to cleave fibrinogen, it was also found to be able to specifically cleave the crown of the V3 loop of human immunodeficiency virus-1 (HIV-1) and promote HIV-1 In addition, the up-regulation of thrombin and the expression of thrombin receptors on the cell surface may be related to the occurrence of HIV-1-related encephalopathy, and thrombin plays a key role in many diseases of the coagulation system. Overexpression of thrombin can lead to thrombosis, but underexpression can ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573G01N33/533
CPCG01N33/533G01N33/573
Inventor 高力李琴邓泽斌时海霞李娆琪陈克平张春霞周阳严丽荣
Owner 镇江永晨科技有限公司
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