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A biosensor based on nucleic acid aptamer, preparation method and application

A biosensor and nucleic acid aptamer technology, applied in the field of biochemistry, can solve the problems of easy photobleaching, short fluorescence lifetime, easy to be interfered, etc., and achieve high target specificity, high detection accuracy, and easy miniaturization. Effect

Active Publication Date: 2016-08-17
ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, optical aptamer sensors also have disadvantages such as short fluorescence lifetime, prone to photobleaching and interference.

Method used

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  • A biosensor based on nucleic acid aptamer, preparation method and application
  • A biosensor based on nucleic acid aptamer, preparation method and application
  • A biosensor based on nucleic acid aptamer, preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0053] In this example, the nucleic acid aptamer-based biosensor self-assembles on the gold film with octadecylthiol, which is bound to carboxyl-functionalized graphene oxide through intermolecular force, and the carboxyl-functionalized graphene oxide is activated The treatment converts the carboxyl group into an activated ester group, and then forms an amide bond with the amino acid on the nucleic acid aptamer molecule through the activated ester group to immobilize the nucleic acid aptamer on the surface of the carboxyl-functionalized graphene oxide.

[0054] The preparation method of the nucleic acid aptamer-based biosensor in this embodiment comprises the following steps:

[0055] (1) Preparation of PBS buffer solution and electrolyte solution: 4.6548g Na 2 HPO 4 12H 2 O, 0.4g KH 2 PO 4 , 16gNaCl and 0.4g KCl were dissolved in 200mL deionized water, and mixed to obtain a PBS buffer solution (biological buffer solution) with a pH value of 7.4; 4.6548g NaCl was added to ...

Embodiment 2

[0065] In this example, the nucleic acid aptamer-based biosensor self-assembles on the gold film with octadecylthiol, which is bound to carboxyl-functionalized graphene oxide through intermolecular force, and the carboxyl-functionalized graphene oxide is activated The treatment converts the carboxyl group into an activated ester group, and then forms an amide bond with the amino acid on the nucleic acid aptamer molecule through the activated ester group to immobilize the nucleic acid aptamer on the surface of the carboxyl-functionalized graphene oxide.

[0066] The preparation method of the nucleic acid aptamer-based biosensor in this embodiment comprises the following steps:

[0067](1) Preparation of PBS buffer solution and electrolyte solution: 4.6548g Na 2 HPO 4 12H 2 O, 0.4g KH 2 PO 4 , 16gNaCl and 0.4g KCl were dissolved in 200mL deionized water, and mixed to obtain a PBS buffer solution (biological buffer solution) with a pH value of 7.4; 4.6548g NaCl was added to e...

Embodiment 3

[0077] In this example, the nucleic acid aptamer-based biosensor self-assembles on the gold film with octadecylthiol, which is bound to carboxyl-functionalized graphene oxide through intermolecular force, and the carboxyl-functionalized graphene oxide is activated The treatment converts the carboxyl group into an activated ester group, and then forms an amide bond with the amino acid on the nucleic acid aptamer molecule through the activated ester group to immobilize the nucleic acid aptamer on the surface of the carboxyl-functionalized graphene oxide.

[0078] The preparation method of the nucleic acid aptamer-based biosensor in this embodiment comprises the following steps:

[0079] (1) Preparation of PBS buffer solution and electrolyte solution: 4.6548g Na 2 HPO 4 12H 2 O, 0.4g KH 2 PO 4 , 16gNaCl and 0.4g KCl were dissolved in 200mL deionized water, and mixed to obtain a PBS buffer solution (biological buffer solution) with a pH value of 7.4; 4.6548g NaCl was added to ...

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Abstract

The invention discloses a nucleic acid aptamer-based biosensor, a preparation method and an application, belonging to the technical field of biochemistry. The biosensor is self-assembled with octadecylthiol on the gold film, and octadecylthiol is combined with carboxyl-functionalized graphene oxide through intermolecular force, and the carboxyl-functionalized graphene oxide is activated to convert the carboxyl group into an activated ester group. The nucleic acid aptamer is immobilized on the surface of carboxyl-functionalized graphene oxide by activating the ester group and the amino acid on the nucleic acid aptamer molecule to form an amide bond; it has good electrochemical activity and can better fix the nucleic acid aptamer , can be used for the detection of various target proteins, and can be used in fields such as biomedicine, food safety and environmental monitoring. The preparation method of the biosensor is simple and easy to operate, in which NaOH and ClCH 2 COONa converts the hydroxyl and epoxy groups on graphene oxide into carboxyl groups. It has a high content of COOH functional groups, has good electrochemical activity, and can better immobilize nucleic acid aptamers.

Description

technical field [0001] The invention specifically relates to a nucleic acid aptamer-based biosensor, a preparation method and application of the biosensor, and belongs to the technical field of biochemistry. Background technique [0002] As a quasi-two-dimensional layered structure, graphene oxide contains a series of oxygen functional groups on the surface, and a large number of hydroxyl and carboxyl active groups between the layers. Graphene oxide and graphene have similar advantages, such as high electrical conductivity, high electron mobility, high specific surface area, and high transparency. Graphene oxide has strong practicability, can be quickly functionalized by chemical methods, and has high biocompatibility. Because the graphene oxide sheet has a large number of hydrophilic acidic functional groups on its surface, it has good wetting properties and surface activity, so that it can be dispersed in pure water to form a stable colloidal suspension. Studies have fou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/42G01N27/327
Inventor 张治红张肖静马永鹏马闯康萌萌何领好王明花蔡立芳彭东来
Owner ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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