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Preparation method of aptamer biosensor for detecting thrombin

A nucleic acid aptamer and biosensor technology, which is applied in the field of nanomaterials and electroanalytical chemical detection, to achieve the effect of improving performance

Inactive Publication Date: 2014-07-16
EAST CHINA JIAOTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The object of the present invention is to provide a method for the detection of thrombin in view of the low sensitivity of the nucleic acid aptamer sensor for the detection of thrombin and the complicated operation of the method for preparing the sensor. Method for preparing nucleic acid aptamer sensor from hydrotalcite-carbon nanotube composite nanomaterial and using it to detect thrombin

Method used

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  • Preparation method of aptamer biosensor for detecting thrombin
  • Preparation method of aptamer biosensor for detecting thrombin
  • Preparation method of aptamer biosensor for detecting thrombin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Nucleic acid aptamer sensors were prepared from hydrotalcite-carbon nanotube composite nanomaterials.

[0031] (1) Mix 10 mL of solution A and 10 mL of suspension B in a beaker to form suspension C.

[0032] Among them, the concentrations of cobalt nitrate and aluminum nitrate in solution A are 0.02mol / L and 0.02 mol / L respectively, and the molar ratio of cobalt nitrate and aluminum nitrate is 1:1; potassium nitrate, potassium chloride and carbon nano The concentrations of the tubes were 0.1 mol / L, 0.05 mol / L, and 0.5 mg / mL, respectively.

[0033] (2) The clean glassy carbon electrode was electrodeposited in the mixed suspension C at the potential of -0.7 V for 80 s to obtain a glassy carbon electrode modified by hydrotalcite-carbon nanotube composite nanoparticles.

[0034] (3) The hydrotalcite-carbon nanotube modified electrode was sequentially placed in 0.5 μmol / L thrombin aptamer for 12 h and soaked in 0.5 mg / mL bovine serum albumin for 40 min to prepare the nuclei...

Embodiment 2

[0036] Nucleic acid aptamer sensors were prepared from hydrotalcite-carbon nanotube composite nanomaterials.

[0037] (1) Mix 15 mL of solution A and 15 mL of suspension B in a beaker to form suspension C.

[0038] Wherein, the concentrations of cobalt nitrate and aluminum nitrate in solution A are 0.03mol / L and 0.02 mol / L respectively, and the molar ratio of cobalt nitrate and aluminum nitrate is 1.5:1; Potassium nitrate, potassium chloride and carbon nano The concentrations of the tubes were 0.2 mol / L, 0.1 mol / L, and 1.5 mg / mL, respectively.

[0039] (2) The clean glassy carbon electrode was electrodeposited in the mixed suspension C at the potential of -0.8 V for 70 s to obtain a glassy carbon electrode modified by hydrotalcite-carbon nanotube composite nanoparticles.

[0040] (3) The hydrotalcite-carbon nanotube modified electrode was sequentially placed in 1.0 μmol / L thrombin aptamer for 12 h and soaked in 1.0 mg / mL bovine serum albumin for 20 min to prepare the nucleic ...

Embodiment 3

[0042] Nucleic acid aptamer sensors were prepared from hydrotalcite-carbon nanotube composite nanomaterials.

[0043] (1) Mix 10 mL of solution A and 10 mL of suspension B in a beaker to form suspension C.

[0044] Wherein, the concentrations of cobalt nitrate and aluminum nitrate in solution A are 0.03mol / L and 0.01 mol / L respectively, and the molar ratio of cobalt nitrate and aluminum nitrate is 3:1; Potassium nitrate, potassium chloride and carbon nano The concentrations of the tubes were 0.3 mol / L, 0.1 mol / L, and 1.0 mg / mL, respectively.

[0045] (2) The clean glassy carbon electrode was electrodeposited in the mixed suspension C at the potential of -0.9 V for 60 s to obtain a glassy carbon electrode modified by hydrotalcite-carbon nanotube composite nanoparticles.

[0046](3) The hydrotalcite-carbon nanotube modified electrode was sequentially placed in 1.0 μmol / L thrombin aptamer for 12 h and soaked in 1.0 mg / mL bovine serum albumin for 40 min to prepare the nucleic aci...

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Abstract

The invention relates to a preparation method of an aptamer biosensor for detecting thrombin. The method is characterized in that a hydrotalcite-carbon nanotube composite nano material modified electrode is prepared through a simple one-step electrodeposition method, and then the aptamer is loaded onto the surface of the hydrotalcite-carbon nanotube composite nano material modified electrode by adopting a strong static adsorption effect to prepare an aptamer modified electrode. By utilizing the synergistic effect of the hydrotalcite-carbon nanotube composite nano material and a specificity binding effect of the aptamer on a substrate, the thrombin can be sensitively, accurately and specifically detected. Other substrate molecules can be detected only by changing the sequence of the aptamer. The aptamer biosensor is suitable for determining the thrombin by utilizing the electrochemical impedance method. The appended drawing is the impedance response drawing of the aptamer biosensor for the thrombin.

Description

technical field [0001] The present invention relates to a method for preparing a nucleic acid aptamer biosensor for detecting thrombin, in particular to a method for preparing a nucleic acid aptamer biosensor for detecting thrombin by using a hydrotalcite-carbon nanotube composite nanomaterial , which belongs to the technical field of nanomaterials and electroanalytical chemical detection. Background technique [0002] Thrombin is an indispensable physiological protease in the human body. It has the effect of rapid hemostasis and can change the fibrinogen in plasma from soluble to insoluble. It plays a key role in pathological and physiological processes such as wound healing and coagulation, so the determination of thrombin has very important practical significance in clinical application and pathological analysis. In recent years, the electrochemical nucleic acid aptamer sensor for detecting thrombin has been favored by many researchers because of its simplicity, rapidi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26G01N33/573
Inventor 杨绍明陈爱喜李红李瑞琴尚培玲
Owner EAST CHINA JIAOTONG UNIVERSITY
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