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Aptamer electrochemical biosensor, as well as preparation method and application thereof

A biosensor and aptamer technology, applied in the field of biosensors, can solve problems such as simulating enzyme activity that are rarely studied, and achieve the effects of low cost, simple synthesis, and wide linear range

Active Publication Date: 2015-03-04
ANHUI NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There are many related studies devoted to the use of DNA-templated metal nanomaterials (such as: gold nanoclusters (AuNCs), silver nanoclusters (AgNCs)) as the detection signal of DNA biosensors, but most of them are Using the optical properties of metal nanoclusters, few studies have explored the use of metal nanoclusters to mimic enzyme activities.

Method used

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  • Aptamer electrochemical biosensor, as well as preparation method and application thereof
  • Aptamer electrochemical biosensor, as well as preparation method and application thereof
  • Aptamer electrochemical biosensor, as well as preparation method and application thereof

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Embodiment Construction

[0052] The present invention will be described in detail below according to the accompanying drawings, which is a preferred embodiment among various implementations of the present invention.

[0053]The preparation of an aptamer electrochemical biosensor based on DNA hybridization chain reaction and silver nanoclusters' simulated enzyme catalysis and the application of amplified detection aptamers include the following steps:

[0054] a. Dissolve the purchased DNA sequences (thiolated DNA: S1, aptamer-specific binding DNA: S2, hairpin DNA 1: HP1, hairpin DNA 2: HP2) in PBS (pH 7.4) buffer solution , and store at low temperature for later use.

[0055] b. Polish the gold electrode with 0.3 and 0.5 μm aluminum powder in turn, and then put it into HNO 3 :H 2 O(v / v)=1:1 solution, ethanol solution, and ultrapure water were used for ultrasonic cleaning, and the ultrasonic cleaning time was 3 to 5 minutes respectively.

[0056] c. Drop-coat the buffer solution containing probes S1...

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Abstract

The invention relates to an aptamer electrochemical biosensor, as well as a preparation method and application therof. The preparation method comprises the following steps: (1) respectively dissolving DNA sequences into a PBS buffer solution; (2) firstly polishing and washing a gold electrode; (3) dispensing the buffer solution containing a probe S1 and a probe S2 onto the gold electrode for culturing; (4) dispensing the PBS buffer solution onto an S2-S1 / GE electrode for culturing; (5) placing S2-S2 / GE into the PBS buffer solution to be cultured; washing the electrode to obtain a Aptasensor; (6) culturing the acquired Aptasensor in the buffer solution, and thus obtaining a decorative electrode HP2-HP1-Aptasensor of a super-sandwiched structure by virtue of DNA hybrid chain reaction; and (7) culturing the HP2-HP1-Aptasensor in the buffer solution to obtain a silver nano-cluster decorated electrode AgNCs / HP2-HP1-Aptasensor. According to the preparation method of the biosensor, the used nano material is simple to synthesize, the energy consumption is low, the cost is low, the biological compatibility is good, the mimic enzyme catalytic activity of the nano material is studied and used as an electrochemical detection signal; moreover, a signal detected by the prepared biosensor is amplified by utilizing the DNA hybrid chain reaction.

Description

technical field [0001] The invention relates to the technical field of biosensors, in particular to assembling silver nano-clusters with simulated enzyme activity on electrochemical biosensors by DNA hybridization chain reaction and using the sensors to amplify and detect target aptamer lysozyme. Background technique [0002] In recent years, the study of DNA is an extremely important aspect in life science research, and the study of DNA biosensors has become a hot spot. There are many related studies devoted to the use of DNA-templated metal nanomaterials (such as: gold nanoclusters (AuNCs), silver nanoclusters (AgNCs)) as the detection signal of DNA biosensors, but most of them are Using the optical properties of metal nanoclusters, few studies have explored the use of metal nanoclusters to mimic enzyme activities. Among the strategies for amplifying the detection signal of DNA biosensors, DNA hybridization chain reaction can be widely used as a good signal amplification ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/26
Inventor 王广凤陈玲
Owner ANHUI NORMAL UNIV
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