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Biosensor based on aptamer and manufacturing method and application thereof

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

Active Publication Date: 2014-01-22
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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  • Biosensor based on aptamer and manufacturing method and application thereof
  • Biosensor based on aptamer and manufacturing method and application thereof
  • Biosensor based on aptamer and manufacturing method and application thereof

Examples

Experimental program
Comparison scheme
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 , 16g NaCl and 0.4g KCl were dissolved in 200mL deionized water, and mixed to obtain a PBS buffer (biological buffer solution) with a pH value of 7.4; add 4.6548g Na to every 200ml of w...

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 biosensor based on an aptamer and a manufacturing method and application thereof and belongs to the field of biochemical technique. According to the biosensor based on the aptamer, n-octadecyl mercaptan is automatically assembled on a gold film, the n-octadecyl mercaptan is combined with carboxyl group graphene oxide through intermolecular force, activating treatment is carried out on the carboxyl group graphene oxide to enable carboxyl to be converted into an active ester group, and an amido bond is formed through the active ester group and amino acid on molecule of the aptamer to enable the aptamer to be fixed on the surface of the carboxyl group graphene oxide. According to the biosensor based on the aptamer, the electrochemical activity is good, the aptamer can be well fixed, and the biosensor can be used for detection of various kinds of target protein and can be applied to the fields such as biomedicine, food safety and environmental monitoring. The manufacturing method of the biosensor based on the aptamer is simple and easy to operate. Due to the fact that hydroxyl and epoxy group which are on the graphene oxide are converted into the carboxyl due to the adoption of NaOH and ClCH2COONa, the COOH functional group content is high, the good electrochemical activity is achieved, and the aptamer can be well fixed.

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 Applications(China)
IPC IPC(8): G01N27/42G01N27/327
Inventor 张治红闫福丰康萌萌杨光彭东来王培远田志壮
Owner ZHENGZHOU UNIVERSITY OF LIGHT INDUSTRY
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